方药用于代谢性肝病的一区基础研究：机制与因果链完整、通路抑制与回补实验、靶点关联验证

Metabolic dysfunction-associated steatotic liver disease (MASLD), formerly known as nonalcoholic fatty liver disease, continues to rise with rapid economic development and poses significant challenges to human health. No effective drugs are clinically approved. MASLD is regarded as a multifaceted pathological process encompassing aberrant lipid metabolism, insulin resistance, inflammation, gut microbiota imbalance, apoptosis, fibrosis, and cirrhosis. In recent decades, herbal medicines have gained increasing attention as potential therapeutic agents for the prevention and treatment of MASLD, due to their good tolerance, high efficacy, and low toxicity. In this review, we summarize the pathological mechanisms of MASLD; emphasis is placed on the anti-MASLD mechanisms of Chinese herbal formula (CHF), especially their effects on improving lipid metabolism, inflammation, intestinal flora, and fibrosis. Our goal is to better understand the pharmacological mechanisms of CHF to inform research on the development of new drugs for the treatment of MASLD.

The prevalence of metabolic dysfunction-associated steatotic liver disease (MASLD) has increased significantly in recent decades and is projected to increase further due to the rising obesity rates. MASLD patients are at higher risk of developing advanced liver diseases “cirrhosis and hepatocellular carcinoma” as well as liver- or cardiovascular-related mortality. Existing lipid-lowering therapies failed to reduce the risk of mortality in these patients. Therefore, there is an urgent need for pharmacotherapies that can control and even reverse this disease. Fanlian Huazhuo Formula (FLHZF) is a combination herbal preparation, and its various individual constituents regulate hepatic lipid metabolism, adipose tissue inflammation, and gut microbiota. Despite, these useful effects, limited information is available on its benefits in diet-induced hepatosteatosis. In this article, we discuss the research findings recently published about the therapeutic effects of FLHZF in suppressing MASLD development and underlying mechanisms. Utilizing a series of in vitro and in vivo experiments, the authors demonstrated for the first time that FLHZF suppresses MASLD in male mice possibly by inhibiting hepatic de novo lipogenesis pathways and reducing hepatocyte death. This study paves the way for future investigations aimed at investigating FLHZF’s role in inhibiting lipogenesis particularly using radioactively-labeled glucose and acetate, and governing hepatocyte mitochondrial function, gut microbiome profile, and its effects in other models of MASLD, and female mice.

The TiaoGanXiaoZhi formula (TGXZ), a traditional Chinese medicine, has been shown to alleviate the progression of metabolic‐associated fatty liver disease (MAFLD) clinically. However, its underlying mechanism remains unclear. This study aimed to investigate the effects and mechanisms of TGXZ in treating MAFLD in mouse models.The MAFLD mouse model was established using a high‐fat diet and 5% fructose water over 16 weeks. At Week 8, mice exhibited significant steatosis, inflammation, and insulin resistance. A total of 42 mice were divided into the normal feed diet (NFD) group (n = 18) and the high‐fat feed diet (HFD) group (n = 24). Six mice from each group were killed at Week 8 for serological and histopathological assessments. The remaining mice were allocated into NFD (n = 6), HFD (n = 6), HFD + TGXZ (n = 6), NFD + TGXZ (n = 6), and HFD + Placebo (n = 6) groups. TGXZ (or placebo) was administered at a clinical equivalent dose of 7.699 g/(kg·d) to the respective groups, while NFD and HFD groups received distilled water. Daily gavage started in Week 9. At Week 16, after fasting, body weight and liver condition were recorded, and mice were euthanized with pentobarbital sodium. Liver tissue was collected for further analysis, and the remaining tissue and feces were stored at −80°C. Data were graphed using GraphPad Prism 8.0.0 and analyzed with SPSS Statistics 25.0. Results are expressed as mean ± standard deviation. Statistical comparisons were made using Student's t‐test for two groups and one‐way ANOVA for more than two groups, with significance set at p < 0.05.Compared to the MAFLD mouse model group, TGXZ treatment significantly downregulated the weight of white adipose tissue (1.61 ± 0.66 vs. 3.06 ± 0.34 g, p < 0.0010), liver weight (1.22 ± 0.16 vs. 1.98 ± 0.39 g, p = 0.0031), and the levels of alanine aminotransferase (20.6 ± 3.4 vs. 46.1 ± 12.3 U/L, p < 0.0010), aspartate aminotransferase (99.9 ± 19.1 vs. 168.4 ± 34.3 U/L, p = 0.0014), cholesterol (2.95 ± 0.56 vs. 4.38 ± 0.34 mmol/L, p = 0.0053), triglycerides (2.25 ± 0.41 vs. 4.18 ± 0.67 mmol/L, p < 0.0010), low‐density lipoprotein (0.66 ± 0.11 vs. 1.41 ± 0.52 mmol/L, p = 0.0073), and total bile acid (0.71 ± 0.41 vs. 2.18 ± 0.61 mmol/L, p = 0.0017), except for high‐density lipoprotein (2.41 ± 0.81 vs. 2.55 ± 0.31 mmol/L, p = 0.5655). The liver transcriptome, fecal microbiota sequencing, and fecal lipidomics analysis demonstrated that TGXZ treatment improved the expression of genes related to lipid metabolism, alleviated intestinal microbiota disorders, and mitigated lipid disturbances caused by MAFLD.Our study demonstrated that TGXZ treatment effectively alleviated the progression of MAFLD. The inhibitory effects of TGXZ on MAFLD may be attributed to its regulation of gut microbiota, lipid metabolism, and hepatic inflammation.

Objective As a metabolic disease, one important feature of non-alcoholic fatty liver disease (NAFLD) is the disturbance of the intestinal flora. Spleen-strengthening and liver-draining formula (SLF) is a formula formed according to the theory of “One Qi Circulation” (Qing Dynasty, 1749) of Traditional Chinese Medicine (TCM), which has shown significant therapeutic effect in patients with NAFLD in a preliminary clinical observation. In this study, we aim to explore the mechanism of SLF against NAFLD, especially its effect on glucolipid metabolism, from the perspective of intestinal flora. Methods A prospective, randomized, controlled clinical study was designed to observe the efficacy and safety of SLF in the treatment of NAFLD. The study participants were randomly and evenly divided into control group and treatment group (SLF group). The control group made lifestyle adjustments, while the SLF group was treated with SLF on top of the control group. Both groups were participated in the study for 12 consecutive weeks. Furthermore, the feces of the two groups were collected before and after treatment. The intestinal flora of each group and healthy control (HC) were detected utilizing 16S rRNA gene sequencing. Results Compared with the control group, the SLF group showed significant improvements in liver function, controlled attenuation parameter (CAP), and liver stiffness measurement (LSM), meanwhile, patients had significantly lower lipid and homeostasis model assessment of insulin resistance (HOMA-IR) with better security. Intestinal flora 16S rRNA gene sequencing results indicated reduced flora diversity and altered species abundance in patients with NAFLD. At the phylum level, Desulfobacterota levels were reduced. Although Firmicutes and Bacteroidetes did not differ significantly between HC and NAFLD, when grouped by alanine transaminase (ALT) and aspartate transaminase (AST) levels in NAFLD, Firmicutes levels were significantly higher in patients with ALT or AST abnormalities, while Bacteroidetes was significantly lower. Clinical correlation analysis showed that Firmicutes positively correlated with gender, age, ALT, AST, LSM, and Fibroscan-AST (FAST) score, while the opposite was true for Bacteroidetes. At the genus level, the levels of Alistipes, Bilophila, Butyricimonas, Coprococcus, Lachnospiraceae_NK4A136 group Phascolarctobacterium, Ruminococcus, UCG-002, and UCG-003 were reduced, whereas abundance of Tyzzerella increased. There was no statistically significant difference in Firmicutes and Bacteroidota levels in the SLF group before and after treatment, but both bacteria tended to retrace. At the genus level, Coprococcus (Lachnospiraceae family), Lachnospiraceae_NK4A136 group (Lachnospiraceae family), and Ruminococcus (Ruminococcaceae family) were significantly higher in the SLF group after treatment, and there was also a tendency for Bilophila (Desulfovibrionaceae family) to be back-regulated toward HC. Conclusions SLF can improve liver function and glucolipid metabolism in patients with NAFLD and lower down liver fat content to some extent. SLF could be carried out by regulating the disturbance of intestinal flora, especially Coprococcus, Lachnospiraceae_NK4A136 group, and Ruminococcus genus.

Objective This study aimed to evaluate the therapeutic effect of the traditional Chinese medicine compound Huatan Qushi Huoxue prescription (HQHF) on Metabolic Associated Fatty Liver Disease (MAFLD) and explore its underlying mechanisms through a multi-method approach. Methods A high-fat diet-induced MAFLD mouse model was established to assess HQHF’s effects on liver pathology, biochemical markers, and oxidative stress. The bioactive components and potential targets of HQHF were identified through network pharmacology. MAFLD-related signaling pathways were explored by integrating data from the Gene Expression Omnibus (GEO) database, followed by molecular docking analyses. For experimental validation, key inflammatory cytokines and proteins (Interleukin-6 (IL-6), Interleukin-1α (IL-1α), Interleukin-1β (IL-1β), Matrix Metallopeptidase 9 (MMP9), Chemokine Ligand 2 (CCL2), and Intercellular Adhesion Molecule 1 (ICAM1)) were measured using Enzyme-Linked Immunosorbent Assay (ELISA), immunohistochemistry (IHC), and immunofluorescence. Results Histopathological analysis showed that HQHF significantly alleviated liver steatosis and inflammatory cell infiltration. Biochemical analysis indicated that HQHF reduced serum and liver triglyceride levels, serum total cholesterol, liver enzymes (Aspartate Aminotransferase, AST, and Alanine Aminotransferase, ALT), and malondialdehyde (MDA), while increasing glutathione (GSH) levels. Network pharmacology identified 33 bioactive components and 89 key targets, which were enriched in pathways including Mitogen-Activated Protein Kinase (MAPK), Toll-like Receptor (TLR), Interleukin-17 (IL-17), and Advanced Glycation End product-Receptor for Advanced Glycation End product (AGE-RAGE). Six core genes (MMP9, IL1α, CCL2, IL1β, IL6, ICAM-1) were highlighted. Molecular docking confirmed strong binding of major HQHF compounds (e.g., quercetin, baicalin) to these targets, and experimental assays further validated their downregulation in HQHF-treated mice. Conclusion HQHF exerts lipid-lowering and anti-inflammatory effects in MAFLD by regulating multiple targets and pathways related to lipid metabolism and inflammation.

BACKGROUND Non-alcoholic fatty liver disease (NAFLD) has become a prevalent issue and a consequence of metabolic syndrome impact on human health. Both of anti-atherosclerosis and anti-hepatic fibrosis capabilities of herbal medicine Ger-Gen-Chyn-Lian-Tang (GGCLT) has attracted attention, but their molecular regulatory mechanisms in a NAFLD model have not been elucidated. The aim of the present study was to explore the bioactivity of db/db mice following treatment with GGCLT. METHODS NAFLD phenotype of db/db mice were treated with GGCLT and lipogenesis, mitochondria dysfunction, mitophagy, macrophage polarization and adipose tissue browning were then evaluated using qRT-PCR and/or Western blot analysis, immunofluorescence, and immunohistochemistry assays, respectively. RESULTS GGCLT not only decreased serum levels of TG and free fatty acids, but glucose and insulin tolerance test in db/db mice. In parallel, GGCLT reduced lipogenesis and hypoxia-inflammation cascades in NAFLD progression. GGCLT reduced lipid accumulation and was accompanied by the enhanced mitochondria biogenesis, M2 macrophage, and decreased M1 macrophage. The latter two events contributing to the anti-inflammation are resulting from mitochondria dynamics, and the lipotoxicity lowering effect of GGCLT of NAFLD mice is mediated by promoting mitophagy in Parkin-dependent and -independent pathways, by mitochondrial fusion over fission manner. GGCLT also inactivated lipogenesis and decreased lipid accumulation in epididymal white adipose tissue with a higher M2/M1 macrophage ratio. CONCLUSIONS Besides in the liver, modulating of mitochondrial biogenesis and adipose tissue browning were characterized by increased Tmem26, Tfam, and Prdm16 expression by GGCLT in EWAT also contributes to the beneficial action in NAFLD.

Non-alcoholic fatty liver disease (NAFLD) has emerged as a global public health concern, affecting over one-quarter of the global population. It is closely associated with the prevalence of obesity and metabolic syndrome. Current treatment options for NAFLD are limited and often have side effects. Traditional Chinese medicine (TCM) offers a promising alternative with its holistic approach and use of multi-component herbal formulations. A recent study explored the potential of the TCM formula, "Fanlian Huazhuo Formula (FLHZF)", to alleviate high-fat diet-induced NAFLD by regulating autophagy and lipid synthesis signaling pathways. TCM has shown advantages in the prevention and treatment of NAFLD due to its efficacy and minimal side effects. However, the complex multicomponent and multitarget characteristics of formulas such as FLHZF present challenges in research. Future studies should focus on utilizing modern techniques to deepen our understanding of the mechanisms of action and active ingredients of Chinese herbal medicines, thereby promoting their modernization.

ABSTRACT Obesity is a complex metabolic disorder characterized by excessive fat accumulation and is closely associated with non‐alcoholic fatty liver disease (NAFLD), a condition that increases the risk of metabolic complications such as insulin resistance, type 2 diabetes, and cardiovascular diseases. JI017 is a recently optimized multi‐herbal formula composed of Angelica gigas, Zingiber officinale , and processed Aconitum carmichaeli. Clinical records of these three herbs indicate their potentially synergistic anti‐obesity effects. This study was conducted to verify the effects of JI017 in an animal model of obesity‐associated NAFLD. The effect of JI017 on glucose metabolism, white adipose tissue (WAT) adipogenesis, brown adipose tissue (BAT) thermogenesis, liver oxidative stress, and gut microbiota composition was assessed. The results showed that JI017 improved glucose sensitivity, reduced WAT mass by suppressing PPARγ and C/EBPα expression, and increased BAT thermogenesis through upregulation of UCP1 and PGC‐1α. Additionally, JI017 reduced liver oxidative stress by increasing HO‐1 and NRF2 expression and modulated the gut microbiota by restoring the Firmicutes/Bacteroidetes ratio. These findings on the multi‐targeted effects of JI017 suggest its potential as a promising therapeutic approach for metabolic diseases including NAFLD.

BACKGROUND Metabolic dysfunction-associated steatotic liver disease (MASLD) prevalence is rising, increasing risks of cirrhosis and hepatocellular carcinoma. The herbal formula Jiangtang Tiaozhi (JTTZF) clinically alleviates MASLD and hepatic steatosis. PURPOSE This study aimed to investigate the therapeutic effects and underlying mechanisms of JTTZF in treating MASLD via the crosstalk between peroxisomes and lipid droplets in hepatocytes. METHODS C57BL/6 J mice on a high-fat diet (HFD) modeling MASLD received daily JTTZF or water for 12 weeks. Serum biomarkers, histology, and liver multi-omics analyses assessed metabolic status, steatosis, targets, and pathways. Oleic acid/palmitic acid-induced HepG2 cells treated with JTTZF-containing serum or ABCD2 overexpression. Mass spectrometry identified JTTZF bioavailable components. Molecular techniques evaluated effects on β-oxidation enzymes, lipid droplet proteins, peroxisomal proteins, and oxidative stress. RESULTS JTTZF significantly improved glucose and lipid metabolism and hepatic steatosis in HFD-fed mice and reduced lipid droplets in liver and in HepG2 cells. DHE staining and ROS flow cytometry confirmed that JTTZF alleviated ROS levels. Multi-omics and validation using RT-PCR, western blotting, and immunohistochemistry revealed that JTTZF regulated enzymes for very long-chain fatty acid metabolism and fatty acid metabolism reprogramming in hepatocellular. Gene overexpression validated JTTZF downregulated the peroxisomal ABCD2 transporter. Mechanistically, JTTZF enhanced lipid droplet metabolism and reduced ROS via the ABCD2/PEX2/ATGL axis, ameliorating insulin resistance and MASLD. CONCLUSIONS This study establishes a hepatocellular fatty acid metabolic reprogramming network in MASLD. Targeting the ABCD2/PEX2/ATGL axis represents a promising therapeutic strategy, with JTTZF showing potential as a therapeutic agent for MASLD.

With the prevalence of obesity and other components of metabolic syndrome, Non-alcoholic fatty liver disease (NAFLD) has become increasingly common. In recent years, much attention has been paid to various plant sources, hoping to find a treatment for NAFLD in plants. The Livsooth authentic herbal formula (LAH, 樂悠本草), a botanical drug formula combined with Puerariae lobatae radix, Lonicerae japonicae flos, Hoveniae semen, and Siraitiae fructus. This study used a network pharmacology approach to predict the potential mechanisms of LAH against NAFLD. Gene Ontology (GO) and KEGG pathway enrichment analyses have identified potential biochemical and signaling pathways. Subsequently, the potential mechanism of action of LAH on NAFLD predicted by network pharmacology analysis was validated in a high-fat diet (HFD)-induced NAFLD model in C57BL/6 mice. Our results demonstrated that LAH ameliorated hepatocyte steatosis in liver tissue by activating the AMPK pathway and decreasing serum triglycerides, low-density lipoprotein, glucose, and cholesterol. Besides, LAH increased the hepatic antioxidant enzymes activities, suggested that LAH improved oxidative stress markers in HFD induced NAFLD mice. In vitro experiments confirmed that the active component of LAH, puerarin, regulates lipid accumulation through the AMPK pathway. In conclusion, our study shows that network pharmacology predictions are consistent with experimental validation. LAH can be a candidate supplement for the prevention of NAFLD.

Introduction Alcoholic liver disease (ALD) is a pathological state of the liver caused by longterm alcohol consumption. Recent studies have shown that the modulation of the gut microbiota and its metabolic products, specifically the short-chain fatty acids (SCFAs), exert a critical role in the evolution and progression of ALD. The Liuweizhiji Gegen-Sangshen beverage (LGS), as a functional beverage in China, is derived from a traditional Chinese herbal formula and has been clinically applied for ALD treatment, demonstrating significant efficacy. However, the underlying mechanisms of LGS for alleviating ALD involving gut microbiota regulation remain unknown. Methods In this study, an ALD murine model based on the National Institute on Alcohol Abuse and Alcoholism (NIAAA) method was established. Results The results showed that oral LGS treatment dose-dependently alleviated alcoholinduced liver injury and inflammation in mice through decreasing levels of ALT, AST and proinflammatory cytokines (TNF-α, IL-6, IL-1β). LGS significantly improved liver steatosis, enhanced activities of alcohol metabolizing enzymes (ALDH and ADH), and reduced the CYP2E1 activity. Notably, regarding most detected indices, the effect of LGS (particularly at medium and high dose) was comparable to the positive drug MTDX. Moreover, LGS had a favorable effect on maintaining intestinal barrier function through reducing epithelial injury and increasing expression of occludin. 16S rRNA sequencing results showed that LGS remarkably modulated gut microbiota structure in ALD mice via recovering alcohol-induced microbial changes and specifically mediating enrichment of several bacterial genera (Alloprevotella, Monoglobus, Erysipelatoclostridium Parasutterella, Harryflintia and unclassified_c_Clostridia). Further study revealed that LGS increased production of SCFAs of hexanoic acid in cecum, promoted alcohol-mediated reduction of GRP43 expression in ileum, and increased serum GLP-1 level. Discussion Overall, LGS exerts a remarkable protective effect on ALD mice through the gut microbiota mediated specific hexanoic acid production and GPR43/GLP-1 pathway.

No abstract available

OBJECTIVE Metabolic syndrome is a complex medical condition that has become an alarming epidemic, but an effective therapy for this disease is still lacking. The use of the herbal formula Huangqisan (HQS) to treat diabetes is documented in the Chinese medical literature as early as 1117 A.D.; however, its therapeutic effects and underlying mechanisms remain elusive. METHODS To investigate the beneficial effects of HQS on metabolic disorders, high-fat diet-induced obesity (DIO), leptin receptor dysfunction (db/db) and low-density lipoprotein receptor-knockout (LDLR-/-) mice were used. Obese mice were treated with either HQS or vehicle. Blood, liver tissue, white fat tissue and brown adipose tissue were harvested at the end of the treatment. Metabolic disease-related parameters were evaluated to test effects of HQS against diabetes, obesity and hyperlipidemia. Aortic arches from LDLR-/- mice were analyzed to investigate the effects of HQS on atherosclerosis. RNA-sequence, quantitative real-time polymerase chain reaction and Western blot were performed to investigate the mechanisms of HQS against metabolic disorder. RESULTS HQS lowered body weight, fasting blood glucose and serum lipid levels and improved glucose tolerance and insulin sensitivity in DIO mice and db/db mice (P < 0.05). HQS also blocked atherosclerotic plaque formation in LDLR-/- mice. HQS suppressed de novo lipid synthesis by reducing the expression of messenger RNA for sterol regulatory element-binding factor 1, stearyl coenzyme A desaturase 1 and fatty acid synthase, and enhancing adenosine 5'-monophosphate-activated protein kinase signaling in both in vivo and in vitro experiments, indicating potential mechanisms for HQS's activity against diabetes. CONCLUSION HQS is effective for reversing metabolic disorder and has the potential to be used as therapy for metabolic syndrome.

Postmenopausal women have an increased risk of developing nonalcoholic fatty liver disease (NAFLD). We formulated a combination of three herb mixtures (HPC03) and observed lipid-lowering efficacy. HepG2 cells were treated with oleic acid to induce an NAFLD model (in vitro). Also, we investigated potential of HPC03 in an ovariectomize- (OVX-) induced NAFLD model (in vivo). We separated the mice into six groups, as follows: SHAM, OVX, OVX + β-estradiol, and OVX + HPC03 (50, 100, and 200 mg/kg). Rats were administered with/without HPC03 for 12 weeks. HPC03 dose dependently inhibited the lipid accumulation involved in lipogenesis in HepG2 cells. The body weight, fat mass, and weights of the liver were decreased in the OVX group than that in the other groups. HPC03 had decreased adiposity that was induced by OVX. HPC03 treatment reduced liver lipid deposition and prevented the increase in serum and liver triglyceride export when there was a deficiency in estradiol. HPC03 improves OVX-induced fatty liver and lipid metabolism. These findings suggest that HPC03 from postmenopausal women has a protective effect during NAFLD conditions.

Non-alcoholic fatty liver disease (NAFLD), often accompanied by insulin resistance, obesity, and hyperlipidemia, is a challenging metabolic disorder to treat. Ge-Lian Qi-Shen Decoction, a traditional Chinese herbal formula, has been clinically used to alleviate symptoms associated with NAFLD, but its underlying mechanisms remain unclear. A NAFLD model was established in C57BL/6J mice using a high-fat diet (HFD). The effects of 4-week GQD intervention at different doses on NAFLD-related symptoms were assessed using biochemical analyses, pathological sections, and oral glucose tolerance tests. ELISA and qPCR were employed to investigate the impact of GQD on serum GLP-1 levels and intestinal Gcg gene expression in NAFLD mice. The direct stimulatory effects of GQD on GLP-1 secretion were examined in NCI-H716 cells and HFD-fed mice. UPLC-MS/MS was used to analyze the composition of ileal contents in GQD-treated mice, and the regulatory effects of 24 identified compounds on GLP-1 secretion were evaluated. Additionally, 16S rDNA sequencing, metabolomics and fecal microbiota transplantation were utilized to explore the role of gut microbiota in GQD’s anti-NAFLD effect. GQD improved HFD-induced hepatic steatosis, impaired glucose tolerance, and elevated blood lipid levels in a dose-dependent manner. It increased serum GLP-1 levels, reduced energy intake, and enhanced glucose tolerance in mice. A single dose of GQD directly elevated serum GLP-1 levels in HFD-fed mice and improved glucose tolerance in a GLP-1-dependent manner. In NCI-H716 cells, GQD promoted intracellular calcium influx and GLP-1 release by activating two G-protein-coupled receptors (GPCRs): bitter taste receptors and TGR5. Compounds such as berberine, coptisine, nuciferine, liensinine, higenamine, aurantio-obtusin, and obtusifolin in GQD activated bitter taste receptors, while maslinic acid and cycloastragenol activated TGR5, facilitating GLP-1 secretion. Furthermore, GQD gavage increased the levels of Muribaculaceae and Akkermansia in mouse feces, leading to elevated concentrations of short-chain fatty acids (SCFAs) such as acetate, propionate, butyrate, and valerate. These SCFAs potentially activated fatty acid-related GPCRs, such as GPR41, in the colon, thereby enhancing colonic Gcg expression. FMT experiment showed that gut microbiota can partially mediate the effect of GQD in increasing GLP-1 levels thus alleviating NAFLD. Some alkaloids, anthraquinones, and triterpenoids in GQD can activate GPCRs, including bitter taste receptors and TGR5, in intestinal endocrine cells, promoting GLP-1 secretion. Simultaneously, GQD regulates gut microbiota composition and metabolism, increasing SCFA levels and Gcg gene expression, leading to sustained elevation of GLP-1 levels. These combined effects contribute to the alleviation of NAFLD symptoms.

INTRODUCTION Steatotic Liver Disease (SLD), including Alcohol-Associated Liver Disease (ALD) and Metabolic Dysfunction-Associated Steatotic Liver Disease (MASLD), is one of the most prevalent chronic liver diseases globally, affecting approximately a quarter of the world's population. It poses a significant burden on global health, but effective treatments are limited. The Yindan-Pinggan (YDPG) formula is a modified traditional Chinese herbal remedy intended to treat liver diseases, but its role in SLD is unclear. This study aimed to explore the effects and mechanisms of YDPG against ALD and MASLD in mice. METHODS The efficacy of YDPG was evaluated by measuring body weight and liver index and assessing hepatic steatosis via H&E and Oil Red O staining. Commercial assay kits were used to detect indicators related to liver injury, lipid metabolism, and glucose metabolism. Intraperitoneal insulin and glucose tolerance tests were performed to assess insulin homeostasis. RT-qPCR, ELISA, and Western blot analysis were applied to measure mRNA or protein expression levels. ChIP-qPCR was performed to assess PPARγ recruitment at the promoters of its target genes. RESULTS YDPG markedly ameliorated ALD and MASLD in mice, with multiple mechanisms potentially contributing to its protective effects against SLD, which these mechanisms include: (1) inhibiting oxidative stress to mitigate hepatic injury via activation of Nrf2 signaling; (2) promoting bile salt excretion via activation of the hepatic FXR-BSEP axis; (3) suppressing hepatic lipid accumulation through dual activation of the intestinal FXR-FGF15 pathway in the ALD model; and (4) suppressing de novo lipogenesis via PPARγ-inhibition in the MASLD model. DISCUSSION This research showed that YDPG demonstrated therapeutic potential for SLD, and the underlying mechanism of YDPG against ALD included alleviation of oxidative stress, anti-inflammatory effects, activation of the intestinal FXR-FGF15 axis, and action against MASLD via inhibition of lipogenesis in hepatocytes. These findings strengthened the evidence that YDPG could be a potential treatment strategy for SLD and offered opportunities for therapeutic advances in the field. CONCLUSION YDPG holds potential for the treatment of ALD, likely by activating FXR and Nrf2 signaling in hepatocytes, enhancing fatty acid oxidation through the intestinal FXR-FGF15 axis, and inhibiting hepatic PPARγ signaling, which exerts a hepatic protective effect against MASLD.

Non-alcoholic fatty liver disease (NAFLD) is characterized by hepatic lipid accumulation and metabolic dysregulation, while effective therapeutic options remain limited. Hepatoprotective Formula 919 (HF919), a traditional Chinese herbal formulation, has shown hepatoprotective effects, but its mechanisms in NAFLD remain incompletely understood. To evaluate the effects of HF919 on hepatic steatosis and metabolic disturbance in NAFLD and the possible involvement of AMPK-associated metabolic regulation. Candidate absorbed constituents of HF919 were characterized by UPLC-HRMS and prioritized by in silico screening based on predicted oral bioavailability and drug-likeness. Network pharmacology, public human liver transcriptomic analyses, and molecular docking were integrated to examine disease-relevant candidate targets and pathways. The in vivo effects of HF919 were evaluated in a high-fat diet-induced NAFLD rat model using histological, biochemical, molecular, and immunohistochemical analyses. Fecal short-chain fatty acid (SCFA) profiling was additionally performed as an exploratory metabolic analysis. HF919 alleviated hepatic steatosis, reduced hepatic lipid accumulation, and improved serum and hepatic lipid-related indices in HFD-induced NAFLD rats. HF919 treatment increased hepatic p-AMPK abundance and Ppara expression and reduced the expression of Tlr4, Nfkb2, and Tgfb1 . Hepatic F4/80-positive staining was significantly reduced, accompanied by increased expression of the macrophage-associated genes Mertk and Arg1 . Public liver transcriptomic analyses showed lower AMPK- and fatty-acid-oxidation-associated transcriptional features in NASH and in more severe histopathological categories, supporting the disease relevance of these metabolic alterations. Integration of HF919-predicted targets with disease-related transcriptomic changes identified metabolic and injury-related candidate pathways, while molecular docking showed favorable predicted docking scores for selected ligand–target pairs. Exploratory SCFA analysis showed shifts in several fecal SCFAs toward the control-associated pattern and associations with selected hepatic metabolic and molecular parameters. HF919 ameliorated hepatic steatosis and metabolic abnormalities in HFD-induced NAFLD rats. The accompanying increase in hepatic p-AMPK abundance and Ppara expression, together with disease-context evidence from human liver transcriptomic datasets, supports the possible involvement of AMPK-associated metabolic regulation. Changes in selected injury-related and macrophage-associated markers and fecal SCFA profiles provided additional secondary or exploratory observations. Further mechanistic studies are needed to establish the causal contribution of AMPK signaling to the effects of HF919.

BACKGROUND Worldwide, non-alcoholic fatty liver disease (NAFLD) is a common chronic liver disease closely associated with obesity, diabetes and other metabolic diseases. Shenling Baizhu powder (SLBZP), a formulation of a variety of natural medicinal plants, has hepatoprotective properties and clinical efficacy in treating non-infectious intestinal disease. SLBZP has improved NAFLD symptoms; however, its mechanism of action is unknown. METHODS We established an NAFLD model in rats given a high-fat diet (HFD), administered different interventions and measured serum biochemical indices and inflammatory factors. Liver tissues were stained with hematoxylin and eosin (HE) and oil red O, and colon tissues were analyzed by immunohistochemistry. The expression profiles of liver TLR4 pathway related protein was confirmed by western blotting. Changes in intestinal microbiota composition were analyzed using a 16S rDNA sequencing technique. RESULTS Of note, SLBZP effectively reduced body weight in HFD-fed rats (p < 0.05). Serum biochemical analysis indicated that SLBZP decreased the serum level of total cholesterol (TC) and improved liver function. Additionally, SLBZP decreased the serum level of endotoxin, tumor necrosis factor α (TNF-α), interleukin-1β (IL-β) (p < 0.05), and decreased the expression of TLR4 pathway related protein. Pathological examination showed that SLBZP alleviates hepatic steatosis and repairs colon mucosa. Microbiome analysis revealed that SLBZP improved the abundance of intestinal microbiota. In taxonomy-based analysis, compared with control rats, SLBZP-treated rats showed obvious changes in intestinal microbiota composition. Moreover, SLBZP increased the relative abundance of short-chain fatty acid (SCFA)-producing bacteria, including Bifidobacterium and Anaerostipes. CONCLUSION Taken together, these results suggest that the effects of SLBZP against NAFLD may be related to the increased abundance of beneficial gut microbiota and decreased levels of LPS in the portal vein.

As one of the most common liver diseases, non-alcoholic fatty liver disease (NAFLD) has attracted widespread international attention given its rising incidence. The current mainstay of treatment for NAFLD is controlled diet with controlled exercise. Significantly, the development of NAFLD is proposed to be related to the liver-gut axis. The gut barrier function may be disrupted in the case of gut microbial dysbiosis, which may increase the entry of bacteria, lipopolysaccharides and endotoxins into the liver, causing an inflammatory response, increasing the load on the liver, and triggering lipid deposition, thereby advancing the progression of NAFLD. Simultaneously, liver injury can further alter intestinal permeability, inducing systemic inflammatory response, creating a vicious cycle that may subsequently exacerbate the development of NAFLD. Accordingly, the present review focused on the exploration of the relationship between the liver-gut axis and NAFLD, dissection of the biological mechanism of NAFLD induced by liver-gut axis dysfunction, and comprehensive elucidation of the research progress of Chinese medicine on the regulation of the liver-gut axis for treating NAFLD. It is expected that these findings may provide fresh and insightful evidence for the development of suitable preventive and therapeutic strategies of NAFLD.

Non-alcoholic fatty liver disease (NAFLD) has become one of the most common chronic diseases in the world, and the effective treatment of NAFLD has been listed as a key problem to be solved urgently in contemporary medicine. Polysaccharides in traditional Chinese medicine (TCM) have a wide range of pharmacological activities. A large number of preclinical studies have confirmed that TCM polysaccharides can interfere with the occurrence and development of NAFLD at multiple interrelated levels, such as improving lipid metabolism and insulin resistance, regulating oxidative stress, alleviating immune inflammatory response, and regulating intestinal microbiota, thus showing great potential as a new anti-NAFLD drug. This paper summarizes the prevention and treatment effect and mechanism of TCM polysaccharides on NAFLD, which provides a basis for the application of TCM polysaccharides in plant medicine and modern medicines, and provides a reference for promoting the development and utilization of TCM polysaccharide resources and the research and development of new drugs for NAFLD.

Non‐alcoholic fatty liver disease (NAFLD) is a metabolic liver disease characterized by the accumulation of excessive liver lipids without alcohol‐induced damage. It has emerged as a significant global health issue. A recent research has indicated that dysregulation of lipid decomposition, uptake, production, oxidation, and secretion causes alterations in various types of lipids, leading to organelle dysfunction and metabolic signaling pathway impairment. Scholars propose that turbid toxin is crucial in the pathogenesis of NAFLD. This article elucidates the meaning of turbid toxin in conjunction with the current literature and analyzes the modern pathophysiological mechanism underlying the development of NAFLD, including turbid toxic substance production, related organelle involvement, biological processes, and pathways, ultimately leading to disease outcomes. The relationship between traditional Chinese medicine and the turbid toxin is also explored to establish a foundation for further mechanistic research on NAFLD.

No abstract available

Lipid metabolism disorders (LMD) can cause a series of metabolic diseases, including hyperlipidemia, obesity, non-alcoholic fatty liver disease (NAFLD) and atherosclerosis (AS). Its development is caused by more pathogenic factors, among which intestinal flora dysbiosis is considered to be an important pathogenic mechanism of LMD. In recent years, the research on intestinal flora has made great progress, opening up new perspectives on the occurrence and therapeutic effects of diseases. With its complex composition and wide range of targets, traditional Chinese medicine (TCM) is widely used to prevent and treat LMD. This review takes intestinal flora as a target, elaborates on the scientific connotation of TCM in the treatment of LMD, updates the therapeutic thinking of LMD, and provides a reference for clinical diagnosis and treatment.

Nonalcoholic fatty liver disease (NAFLD) has become the world's largest chronic liver disease, while there is still no specific drug to treat NAFLD. Traditional Chinese Medicine (TCM) have been widely used in hepatic diseases for centuries in Asia, and TCM’s holistic concept and differentiation treatment of NAFLD show their advantages in the treatment of this complex metabolic disease. However, the multi-compounds and multi-targets are big obstacle for the study of TCM. Here, we summarize the pharmacological actions of active ingredients from frequently used single herbs in TCM compounds. The combined mechanism of herbs in TCM compounds are further discussed to explore their comprehensive effects on NAFLD. This article aims to summarize multiple functions and find the common ground for TCM treatment on NAFLD, thus providing enrichment to the scientific connotation of TCM theories and promotes the exploration of TCM therapies on NAFLD.

No abstract available

Non-alcoholic fatty liver disease (NAFLD) is a chronic liver disease with high incidence and is closely related to metabolic syndrome. If not controlled, it may eventually become hepatocellular carcinoma (HCC). Ferroptosis, a non-apoptotic form of programmed cell death (PCD), is closely related to NAFLD and HCC, and the mechanisms of action involved are more complex. Some studies have demonstrated that many drugs inhibit ferroptosis and protect liver steatosis or carcinogenesis. The role of Traditional Chinese Medicine (TCM), especially herbs or herbal extracts, has received increasing attention. However, there are relatively few review articles on the regulation of NAFLD by TCM through ferroptosis pathway. Here, we summarize the TCM intervention mechanism and application affecting NAFLD/NAFLD-HCC via regulation of ferroptosis. This article focuses on the relationship between ferroptosis and NAFLD or NAFLD-HCC and the protective effect of TCM on both by targeting ferroptosis. It not only summarizes the mechanism of early prevention and treatment of NAFLD, but also provides reference ideas for the development of TCM for the treatment of metabolic diseases and liver diseases.

Metabolic syndrome mainly includes obesity, type 2 diabetes (T2DM), alcoholic fatty liver (NAFLD) and cardiovascular diseases. According to the ancient experience philosophy of Yin-Yang, monarch-minister compatibility of traditional Chinese medicine, prescription is given to treat diseases, which has the advantages of small toxic and side effects and quick effect. However, due to the diversity of traditional Chinese medicine ingredients and doubts about the treatment theory of traditional Chinese medicine, the mechanism of traditional Chinese medicine is still in doubt. Gastrointestinal tract is an important part of human environment, and participates in the occurrence and development of diseases. In recent years, more and more TCM researches have made intestinal microbiome a new frontier for understanding and treating diseases. Clinically, nonalcoholic fatty liver disease (NAFLD) and diabetes mellitus (DM) often co-occur. Our aim is to explain the mechanism of interaction between gastrointestinal microbiome and traditional Chinese medicine (TCM) or traditional Chinese medicine formula to treat DM and NAFLD. Traditional Chinese medicine may treat these two diseases by influencing the composition of intestinal microorganisms, regulating the metabolism of intestinal microorganisms and transforming Chinese medicinal compounds.

Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease around the world, and it often coexists with insulin resistance-related diseases including obesity, diabetes, hyperlipidemia, and hypertension, which seriously threatens human health. Better prevention and treatment strategies are required to improve the impact of NAFLD. Although needle biopsy is an effective tool for diagnosing NAFLD, this method is invasive and difficult to perform. Therefore, it is very important to develop more efficient approaches for the early diagnosis of NAFLD. Traditional Chinese medicine (TCM) can play a certain role in improving symptoms and protecting target organs, and its mechanism of action needs to be further studied. Metabolomics, the study of all metabolites that is thought to be most closely associated with the patients’ characters, can provide useful clinically biomarkers that can be applied to NAFLD and may open up new methods for diagnosis. Metabolomics technology is consistent with the overall concept of TCM, and it can also be used as a potential mechanism to explain the effects of TCM by measuring biomarkers by metabolomics. Based on PubMed/MEDLINE and other databases, this paper retrieved relevant literature NAFLD and TCM intervention in NAFLD using metabolomics technology in the past 5 years were searched, and the specific metabolites associated with the development of NAFLD and the potential mechanism of Chinese medicine on improving symptoms were summarized.

As a classical traditional Chinese patent medicine, Shugan Yipi Granule is widely used in China to treat non-alcoholic fatty liver disease (NAFLD) recently. Our previous study confirmed that Shugan Yipi Granule are effective in NAFLD. However, its underlying mechanism is still unknown. This study aims to investigate the mechanism of Shugan Yipi Granule on NAFLD based on network pharmacology prediction, liquid chromatography-mass spectrometry (LC-MS) analysis and in vitro verification. We obtained the active ingredients and targets of Shugan Yipi Granule and NAFLD from 6 traditional Chinese medicine databases, and the crucial components and targets screened by protein-protein interaction (PPI) network were used for molecular docking. Plasma metabolomics of NAFLD patients treated with Shugan Yipi Granule for one month was analyzed using LC-MS methods and MetaboAnalyst 4.0 to obtain significant differential metabolites and pathways. Finally, free fatty acid (FFA) induced HepG2 cells were treated with different concentrations of quercetin and kaempferol, then oil red o (ORO) and triglyceride (TG) level were tested to verify the lipid deposition of the cell. Network pharmacology analysis showed that the main active ingredients of Shugan Yipi Granule include quercetin, kaempferol and other 58 ones, as well as 188 potential targets. PI3K/Akt signaling pathway was found to be the most relevant pathway for the treatment of NAFLD. Non-targeted metabolomics showed that quercetin and kaempferol were significantly up-regulated differential metabolites and were involved in metabolic pathways such as thyroid hormone signaling. In vitro results showed that quercetin, kaempferol were effective in reducing lipid deposition and TG content by inhibiting cellular fatty acid uptake. Ultimately, with the network pharmacology and serum metabolomics analysis, quercetin and kaempferol were found to be the important active ingredients and significantly up-regulated differential metabolites of Shugan Yipi Granule against NAFLD, which we inferred that they may regulate NAFLD through PI3K/Akt signaling pathway and thyroid hormone metabolism pathway. The in vitro experiment verification results showed that quercetin and kaempferol attenuated the lipid accumulation and TG content by inhibiting the fatty acid uptake in the FFA-induced HepG2 cell. Current study provides the necessary experimental basis for subsequent in-depth mechanism research.

BACKGROUND Wuling capsule is a traditional Chinese medicine composed of four herbals. It has been widely used to treat chronic active hepatitis and has shown significant efficacy in hyperlipidemia. However, the treatment of NAFLD disease has not been studied in depth. METHODS Firstly, the potential bioactive compounds in Wuling capsules were identified by TCMSP (https://old.tcmsp-e.com/tcmsp.php). Secondly， the pathway and GO function were analyzed by using the DAVID database (https://david.ncifcrf.gov/). Then, the molecular docking techniques were used to confirm the accuracy of binding between key targets and components. Furthermore, the experimental pharmacology validation was conducted using RT-qPCR and WB of the NAFLD model. RESULTS A total of 138 active compounds and 40 common potential targets associated with NAFLD were identified through network pharmacology. The pathway and functional enrichment analysis showed that the Wuling capsule was associated with the PI3K-AKT and HIF-1α signaling pathways. In vivo experiments showed that the Wuling capsule could reduce IL-6, TNF-α, and HIF-1α proteins and up-regulate STAT3 and VEGFA levels (P < 0.05), thus alleviating liver inflammation. CONCLUSIONS With the support of network pharmacology and animal experiments, the study preliminarily investigated the effect of the Wuling capsule on liver inflammation by regulating the HIF-1α signaling pathway, thereby protecting liver function and treating NAFLD.

BackgroundNonalcoholic fatty liver disease (NAFLD) represents one of the most common forms of liver disease worldwide, and it is always regarded as a consequence of a sedentary, food-abundant lifestyle, sitting for an extended time, and a low physical activity level, which often coincide with chronic and long-lasting psychological stress. A Chinese medicine Sinisan (SNS) may be a potential formula for treating this kind of disease.MethodsIn this study, a long-term chronic restraint stress protocol was used to investigate the mechanism underlying stress-induced NALFD. To investigate the effect of SNS treatment on stress-induced NAFLD, we measured the liver and serum values of total cholesterol (TC), triglyceride (TG), liver free fatty acids (FFA), low-density lipoprotein, superoxide dismutase, tumor necrosis factor-α, malondialdehyde, interleukin (IL)-6, and serum values of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase. Results are shown as a mean ± standard deviation. Significant differences between the groups were evaluated using the Student t-test. For multiple comparisons, one-way analysis of variance (ANOVA) was used. If the results of ANOVA indicated significant differences, post hoc analysis was performed with the Tukey test or Dunnett test, and p < 0.05 was considered statistically significant.ResultsLong-term chronic stress led to steatosis and non-alcoholic steatohepatitis. Additionally, SNS treatment significantly increased body weight gain (p < 0.01) and sucrose preference (p < 0.001), and it reduced the liver values of TC, TG, and FFA (p < 0.05). SNS also reduced the serum values of AST and ALT (p < 0.001), and the liver value of IL-6 (p < 0.01).ConclusionsThis study’s results demonstrate that psychological stress may be a significant risk factor of NAFLD. Furthermore, the traditional Chinese medicine formula SNS may have some beneficial effect in antagonizing psychological stress and stress-related NAFLD.

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The prevalence of nonalcoholic fatty liver disease (NAFLD) is increasing as obesity and diabetes become more common. There are no drugs approved for NAFLD yet. Qige decoction (QGD), a traditional Chinese medicine (TCM) formula, is used for NAFLD and hyperlipidemia treatment in TCM and has shown hypolipidemic and hepatoprotective effects. This study tried to interpret the pharmacology and molecular mechanisms of QGD in NAFLD rats. Firstly, the therapeutic effects of QGD on high-fat diet (HFD)-induced NAFLD rats were evaluated. Then, integration of lipidomics and transcriptomics was conducted to explore the possible pathways and targets of QGD against NAFLD. QGD at low dosage (QGL) administration reduced serum total cholesterol (TC), triglyceride (TG), and low-density lipoprotein cholesterol (LDL-C) (P < 0.05). Liver histopathology indicated that QGL could alleviate hepatic steatosis. The main differential lipids (DELs) affected by QGD were glycerolipids. KEGG enrichment analysis suggested that the main pathways by which QGD improved NAFLD may be cholesterol metabolism, glycerolipid metabolism, and insulin resistance. Transcriptome sequencing identified 179 upregulated and 194 downregulated mRNAs after QGD treatment. An interaction network based on DELs and differential genes (DEGs) suggested that QGD inhibited hepatic steatosis mainly by reducing hepatic insulin resistance and triglyceride biosynthesis via the PPP1R3C/SIK1/CRTC2 and PPP1R3C/SIK1/SREBP1 signal axis, respectively. These findings indicated that QGD could protect against NAFLD induced by HFD. The improvement of hepatic insulin resistance and the reduction of triglyceride biosynthesis might be the potential mechanisms.

Nonalcoholic fatty liver disease (NAFLD) is a clinicopathological syndrome of abnormal lipid deposition in the liver mediated by nonalcohol intake. The Gexia Zhuyu decoction, a classic traditional Chinese medicine compound, is widely used in the clinical treatment of NAFLD. However, its specific efficacy and underlying mechanisms have not been elucidated yet. This study aimed to quantitatively evaluate the efficacy of the Gexia Zhuyu decoction using pharmacodynamics and to explore its molecular mechanisms in conjunction with proteomics. High-fat diets and methionine choline-deficient diets were used to induce various NAFLD progression stages in mouse models. The effects of oral Gexia Zhuyu decoction administration on NAFLD were evaluated by measuring the serum and liver indicators of the treated mice before and after drug intervention and by comparing the changes in liver tissue. Liver TRPM4 mRNA and protein levels were measured using reverse transcription-polymerase chain reaction and Western blotting, respectively. Experimental data showed that serum ALT, AST, and liver triglyceride (TG) levels in each disease stage group of drug intervention mice decreased, and high-density lipoprotein (HDL) and superoxide dismutase (SOD) levels increased. Liver TG levels decreased after drug intervention in the liver fibrosis mice, but serum TG levels increased. Furthermore, cellular fatty changes, inflammatory changes, and fibrous tissue proliferation were all relieved. The TRPM4 protein and mRNA levels in the liver tissue were decreased, and the microRNA (miRNA)-24 expression was increased. The Gexia Zhuyu decoction has a clear therapeutic effect at each stage of NAFLD. It likely acts by altering miRNA-24 expression and regulating the target TRPM4 protein pathway to achieve NAFLD treatment.

Abstract Metabolic dysfunction-associated steatotic liver disease (MASLD) is a chronic liver disease with rising prevalence and disease burden. However, despite recent therapeutic advances, effective and broadly applicable treatment options remain limited, prompting continued efforts to develop novel therapeutic agents. Traditional Chinese Medicine (TCM) has attracted growing interest in MASLD management because its bioactive compounds can target multiple pathogenic processes. However, many TCM-derived compounds are limited by poor solubility, low bioavailability, and insufficient tissue specificity. Nanotechnology-based formulations enable controlled release and targeted delivery, offering a strategy to improve the utilization and therapeutic efficacy of TCM-derived active ingredients against MASLD. Based on a structured literature search across four databases, 45 representative studies were included and narratively synthesized according to nanoplatform type, design features, and mechanism-related therapeutic actions. Compared with previous broader reviews on TCM nanomedicine or MASLD-related nanotherapies, this review particularly emphasizes MASLD-oriented TCM nanoformulations from the perspectives of platform classification, design features, and mechanism-related therapeutic actions. We also discuss current challenges and future directions for clinical translation.

Abstract Metabolic dysfunction–associated steatohepatitis (MASH) is a progressive form of metabolic dysfunction–associated steatotic liver disease (MASLD) characterized by steatosis, inflammation, hepatocellular injury, and fibrosis. The pathogenesis of MASH is complex and involves multiple factors, such as lipotoxicity, insulin resistance (IR), genetic susceptibility, endoplasmic reticulum (ER) stress, mitochondrial dysfunction, and dysregulation of the hepatic immune microenvironment. Currently, the clinical treatment for MASH remains challenging. Traditional Chinese medicine (TCM) has shown broad prospects in the prevention and treatment of MASH owing to its multi-target and multi-pathway regulation. This review summarizes the pathophysiological basis of MASH relevant to TCM intervention and discusses the effects of TCM formulas, active compounds, and comprehensive treatment strategies on lipid metabolism, IR, inflammation, autophagy and ferroptosis, intestinal barrier function, immune regulation, fibrosis, and clinical outcomes. We also discuss current clinical evidence, safety considerations, and remaining limitations, with the aim of providing a clearer basis for the clinical application and further investigation of TCM in MASH management.

No abstract available

Berberine (BBR), the core alkaloid of the traditional Chinese medicine Coptis chinensis (Huanglian), exhibits a notable pharmacological paradox: it possesses an oral bioavailability of less than 1% yet exerts broad therapeutic effects across the full spectrum of liver diseases, from metabolic dysfunction-associated steatotic liver disease (MASLD) through fibrosis to hepatic malignancies. This review constructs an integrated mechanistic framework to interpret BBR's holistic pharmacological characteristics, which distinguish it from conventional single-target drugs. In MASLD, BBR engages a convergent AMPK-SIRT-Nrf2 network to ameliorate insulin resistance, lipid dysregulation, oxidative stress, and hepatic inflammation. Combined with the complementary actions of its bioactive metabolites, it achieves parent-metabolite coordinated efficacy that embodies the holistic principle of traditional Chinese medicine. In liver fibrosis, BBR may block hepatic stellate cell activation via the TGF-β1/Smad3 pathway and eliminate activated HSCs through multiple programmed cell death pathways, particularly ferroptosis, which may overcome apoptosis resistance. In hepatic malignancies, BBR suppresses tumor progression, metastasis, and chemoresistance via apoptotic, ferroptotic, and immunomodulatory mechanisms. Current clinical evidence validates its metabolic benefits in MASLD, whereas clinical data regarding fibrosis and hepatic malignancies remain insufficient. Improving its poor bioavailability through structural modification and nanocarrier delivery is critical for clinical translation. This review provides a modern pharmacological rationale for the clinical application of Coptis chinensis-based formulas against progressive liver diseases.

BACKGROUND Metabolic dysfunction-associated steatotic liver disease (MASLD) is the leading cause of liver diseases worldwide and imposes an increasingly heavy burden on society. After years of research, salvianolic acid B (Sal B), the main component of traditional Chinese medicine Salvia miltiorrhiza, exhibits significant efficacy in alleviating fatty liver; however, its precise mechanism of action against MASLD remains unclear. PURPOSE This study aimed to investigate the underlying pharmacological mechanism of Sal B in treating MASLD, using both in vivo and in vitro models. STUDY DESIGN A systematic pharmacology approach combining multi-species phenotypic screening with proteomics and molecular target verification was employed to elucidate the mechanism of Sal B against MASLD. METHODS MASLD models were established in thioacetamide‑treated zebrafish, MCD diet‑fed mice, and oleate/palmitate‑exposed AML12 hepatocytes. The therapeutic effects of Sal B were assessed by histology, proteomics, Seahorse metabolic analysis, calcium imaging, flow cytometry, and molecular binding assays such As spr, CETSA, and competitive labeling. The involvement of Piezo1 and the C215 residue was examined by siRNA‑mediated knockdown, NEM competition, and expression of wild‑type versus C215S mutant protein. RESULTS Sal B treatment was associated with reduced hepatic lipid accumulation, inflammation, oxidative stress, and energy metabolism disturbances in all three models. Proteomic analysis suggested modulation of a PPAR‑centered metabolic network. Sal B appeared to interact with the Piezo1 extracellular domain, and the C215S mutation markedly decreased this interaction. In Piezo1‑overexpressing HEK293T cells, Sal B attenuated YODA1‑induced Ca²⁺ influx; this effect was not observed following NEM pretreatment or Piezo1 knockdown. Sal B also reduced YODA1‑elicited mitochondrial Ca²⁺ overload and ROS production in AML12 cells, effects that were less evident when Piezo1 was knocked down. Moreover, Piezo1 knockdown alone produced changes in PPAR‑related genes similar to those induced by Sal B, and adding Sal B to knockdown cells did not lead to further alterations. CONCLUSION The available evidence suggests that Sal B can inhibit Piezo1 channel activity, possibly through interaction with cysteine 215 in its extracellular domain, and that this may contribute to alleviation of mitochondrial Ca²⁺ overload, oxidative stress, and metabolic dysregulation in MASLD models. Importantly, the identification of C215 as the binding site is based on computational prediction and SPR assays using the truncated extracellular domain; direct functional confirmation with full‑length C215S mutant protein remains to be performed. These findings point to C215 as a candidate binding residue for Sal B and support further investigation of Piezo1‑targeted strategies for MASLD.

Metabolic dysfunction-associated steatotic liver disease (MASLD), formerly discussed largely under the non-alcoholic fatty liver disease (NAFLD) framework, is now one of the most common chronic liver diseases worldwide. Its progression from simple steatosis to metabolic dysfunction-associated steatohepatitis (MASH), fibrosis, cirrhosis, and hepatocellular carcinoma (HCC) reflects the interaction of lipid overload, insulin resistance, oxidative stress, immune activation, hepatic stellate cell activation, and oncogenic remodeling. Although resmetirom and glucagon-like peptide-1 receptor agonist-based strategies have recently advanced the treatment landscape for selected patients with MASH and fibrosis, many patients remain outside the indications of current drug therapy, and safe adjunctive strategies across earlier disease stages remain an unmet need. Ganzaoning granule is a traditional Chinese medicine formulation derived from the anti-HCC precursor formula Ganfujian and has been used clinically in China for fatty liver disease and related chronic liver conditions. This narrative review summarizes the available evidence on Ganzaoning, including its phytochemical profile, network pharmacology findings, preclinical studies, and clinical observations in MASLD/NASH populations. Existing studies suggest that Ganzaoning or its related active fractions may modulate lipid metabolism through the AMP-activated protein kinase (AMPK)/peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1α) axis, attenuate inflammatory signaling involving nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) and advanced glycation end-product (AGE)-receptor for advanced glycation end-product (RAGE) pathways, and influence fibrosis- and hepatocarcinogenesis-related molecular processes. However, much of the mechanistic evidence remains preclinical or inferred from network pharmacology, and the clinical evidence is limited by single-center designs, short follow-up, composite endpoints, and limited use of histological or advanced imaging outcomes. We therefore position Ganzaoning as a biologically plausible candidate adjunctive therapy rather than an established MASLD treatment. Future research should prioritize standardized product quality control, pharmacokinetic and pharmacodynamic characterization, herb-drug interaction assessment, and multicenter, double-blind, placebo-controlled trials using accepted MASLD endpoints.

ETHNOPHARMACOLOGICAL RELEVANCE Rubia cordifolia L. (known as Qiancao in Chinese herb) is a commonly used herbal medicine in clinical practice. It exhibits tropism to the Liver Meridian and is traditionally recognized for its effects in cooling blood to stop bleeding and invigorating blood circulation to dispel stasis. Alizarin, its main anthraquinone component, has shown potential for clinical application. The nucleotide oligomerization domain-like receptors containing pyrin domain 3 (NLRP3) inflammasome is a promising therapeutic target for metabolic dysfunction-associated steatotic liver disease (MASLD). This study aimed to investigate the efficacy and mechanism of alizarin in treating MASLD via inhibition of the NLRP3 inflammasome. MATERIALS AND METHODS We examined the effect of alizarin on NLRP3 inflammasome activation. The roles of alizarin in regulating NLRP3 transcription and inflammasome assembly were assessed. Its influence on lipid-induced NLRP3 activation was evaluated in mouse primary hepatocytes and Kupffer cells. The therapeutic potential of alizarin in ameliorating MASLD through NLRP3 inhibition was further verified in vivo. RESULTS Alizarin inhibited NLRP3 inflammasome activation by reducing the cleavage of pro-IL-1β and pro-caspase-1 in mouse bone marrow-derived macrophages (BMDMs). Similar inhibitory effects were observed in BMDMs, primary hepatocytes, and Kupffer cells stimulated with LPS plus cholesterol or oleic acid/palmitic acid (OA/PA). Alizarin did not affect LPS-induced phosphorylation of P65, JNK, ERK, or P38. Mechanistically, it suppressed NLRP3 inflammasome assembly by inhibiting the interaction between NLRP3 and ASC. In a methionine-choline deficient (MCD) diet-induced mouse model of MASLD, alizarin improved liver function, reduced serum inflammatory markers, lowered histopathological inflammation scores, and attenuated liver fibrosis, likely through regulation of the NLRP3 inflammasome. CONCLUSIONS Alizarin inhibits NLRP3 inflammasome assembly and may serve as a potential therapeutic agent for MASLD by targeting the NLRP3 inflammasome.

ETHNOPHARMACOLOGICAL RELEVANCE Artemisia capillaris Thunb. (ACT) is a plant in the Asteraceae family. Its traditional effects are to clear away dampness and heat, promote gallbladder and reduce jaundice. Traditional Chinese medicine believes that MASLD is a damp-heat syndrome. The group's previous study showed that Artemisia capillaris Thunb. Water Extract (ACTE) has an improved effect on MASLD. AIM OF THE STUDY AND METHODS In order to further understand its mechanism of action, this study established a mouse MASLD model and a HepG2 cell lipid droplet model, combined small RNA sequencing and miRNA transfection experiments, to explore the mechanism of ACTE to improve MASLD by modulating miRNA-targeted mRNA. Non-targeted metabolomics method was used to detect and analyze ACTE. RESULTS This study screened miR-34a-5p and confirmed its target mRNA-Sirtuin 1 (Sirt1). MASLD induced high expression of miR-34a-5p and low expression of Sirt1, and ACE reversed these changes. When overexpressing miR-34a-5p or knocking down Sirt1, the effect of ACE in reducing PO (palmitic acid and oleic acid complex)-induced lipid accumulation in HepG2 cells was attenuated. ACTE reduces the expression of FASN, SCD1, ACC, and SREBP-1c, promotes the expression of CPT-1 and HSL, thereby reducing lipid accumulation. CONCLUSIONS ACTE activates Sirt1 by inhibiting the expression of miR-34a-5p, thereby reducing liver lipid accumulation and improving HFD-induced MASLD. These findings highlight the potential of ACTE in reducing weight, controlling obesity, and improving lipid metabolism disorders.

Type 2 diabetes mellitus (T2DM) and nonalcoholic fatty liver disease (NAFLD) are the most problematic metabolic diseases in the world. NAFLD encompasses a spectrum of severity, ranging from simple steatosis to non-alcoholic steatohepatitis (NASH) and fibrosis, increasing the risk of cirrhosis and hepatocellular carcinoma. Importantly, NAFLD is closely linked to obesity and tightly interrelated with insulin resistance and T2DM. T2DM and NAFLD (T2DM-NAFLD) are called as the Xike Rixijing Disease and Tonglaga Indigestion Disease respectively, in Mongolian medicine. Xike Rixijing Disease maybe develop into Tonglaga Indigestion Disease. Forturnately many Mongolian medicines show efficient treatment of T2DM-NAFLD, such as Agriophyllum squarrosum, Haliyasu (dried powder of camel placenta), Digeda-4 (herbs of Lomatogonium carinthiacum, rhizomata of Coptis chinensis, ripe fruits of Gardenia jasminoides, herbs of Dianthus superbus), Guangmingyan Siwei Decoction Powder (Halite, ripe fruits of Terminalia chebula, rhizomata of Zingiber officinale, fruit clusters of Piper longum), Tonglaga-5 (ripe fruits of Punica granatum, barks of Cinnamomum cassia, ripe fruits of Amomum kravanh, fruit clusters of Piper longum, flowers of Carthamus tinctorius), Tegexidegeqi (rhizomata of Inula helenium, ripe fruits of Gardenia jasminoides, rhizomata of Platycodon grandiflorum, rhizomata of Coptis chinensis, heartwood of Caesalpinia sappan), Ligan Shiliu Bawei San (ripe fruits of Punica granatum, barks of Cinnamomum cassia, ripe fruits of Amomum kravanh, fruit clusters of Piper longum, flowers of Carthamus tinctorius, ripe fruits of Amomum tsao-ko, rhizomata of Zingiber officinale), etc. Principles of Mongolian medicine in treating diseases: by balancing “three essences or roots” and “seven elements”, strengthening liver and kidney function, transporting nutrients to enhance physical strength and disease resistance, and combined with drugs for comprehensive conditioning treatment. However, their molecular mechanisms remain unclear. In this review, we prospect that Mongolian medicines might be a promising treatment for T2DM-NAFLD by activating P2X7R/NLRP3/NF-κB inflammatory pathway via lipid-sensitive nuclear receptors (i.e., FXR and LXR).

ETHNOPHARMACOLOGICAL RELEVANCE Shugan Xiaozhi (SGXZ) decoction is a traditional Chinese medicine used for treating nonalcoholic steatohepatitis (NASH). It has been used clinically for over 20 years and proved to be effective; however, the molecular mechanism underlying the effects of SGXZ decoction remains unclear. AIM OF THE STUDY We analyzed the chemical components, core targets, and molecular mechanisms of SGXZ decoction to improve NASH through network pharmacology and in vivo experiments. MATERIALS AND METHODS The chemical components, core targets, and related signaling pathways of SGXZ decoction intervention in NASH were predicted using network pharmacology. Molecular docking was performed to verify chemical components and their core targets. The results were validated in the NASH model treated with SGXZ decoction. Mouse liver function was assessed by measuring ALT and AST levels. TC and TG levels were determined to evaluate lipid metabolism, and lipid deposition was assessed via oil red O staining. Mouse liver damage was determined via microscopy following hematoxylin and eosin staining. Liver fibrosis was assessed via Masson staining. Western blot (WB) and immunohistochemical (IHC) analyses were performed to detect inflammation and the expression of apoptosis-related proteins, including IL-1β, IL-6, IL-18, TNF-α, MCP1, p53, FAS, Caspase-8, Caspase-3, Caspase-9, Bax, Bid, Cytochrome c, Bcl-2, and Bcl-XL. In addition, WB and IHC were used to assess protein expression associated with the TLR4/MyD88/NF-κB pathway. RESULTS Quercetin, luteolin, kaempferol, naringenin, and nobiletin in SGXZ decoction were effective chemical components in improving NASH, and TNF-α, IL-6, and IL-1β were the major core targets. Molecular docking indicated that these chemical components and major core targets might interact. KEGG pathway analysis showed that the pathways affected by SGXZ decoction, primarily including apoptosis and TLR4/NF-κB signaling pathways, interfere with NASH. In vivo experiments indicated that SGXZ decoction considerably ameliorated liver damage, fibrosis, and lipid metabolism disorder in MCD-induced NASH mouse models. In addition, WB and IHC verified the underlying molecular mechanisms of SGXZ decoction as predicted via network pharmacology. SGXZ decoction inhibited the activation of apoptosis-related pathways in MCD-induced NASH mice. Moreover, SGXZ decoction suppressed the activation of TLR4/MyD88/NF-κB pathway in MCD-induced NASH mice. CONCLUSION SGXZ decoction can treat NASH through multiple targets and pathways. These findings provide new insights into the effective treatment of NASH using SGXZ decoction.

Nonalcoholic fatty liver disease (NAFLD), particularly the histological phenotype nonalcoholic steatohepatitis (NASH), is considered the primary cause of chronic liver diseases. Shugan Xiaozhi decoction (SX), a composite Chinese medicinal formula, has shown therapeutic potential for NASH, although the underlying mechanisms remain incompletely defined. In this study, the effects and mechanisms of SX in rats with NASH and fibrosis induced by a high-fat and high-cholesterol (HFHC) diet were investigated. NASH rats with fibrosis were induced by an HFHC diet, with different doses of SX administered. Biochemical indices, histological changes, and gene expression were assessed to evaluate lipid metabolism, inflammation, fibrosis of the liver. The histological changes of white and brown adipose tissue were also determined. Hepatic transcriptomics and network pharmacology based on the blood components were employed to elucidate the biological processes and signaling pathways, which was subsequently verified, and the material basis of SX was investigated through docking simulations. SX treatment effectively corrected disordered lipid profiles, mitigated hepatic phagocyte infiltration, and alleviated damage in the white and brown adipose tissues. It also downregulated the expression of NLRP3, caspase-1, IL-1β, α-SMA, Col1a1, and TGF-β, to improve hepatic inflammation and fibrosis. Mechanistically, transcriptomics and network pharmacology analyses revealed that SX ameliorated hepatic oxidative stress and activated the AMPK pathway, and the downstream effector Nrf2 and inactivated of SREBP1/FAS signaling. Three putative phytochemicals, namely, gallic acid, protocatechuic acid, and rhein with the highest binding affinity for AMPKα were also identified. SX ameliorates HFHC diet-induced NASH by regulating hepatic oxidative stress and AMPK pathway, suggesting the potential therapeutic value of SX in NASH treatment.

BACKGROUND Nonalcoholic steatohepatitis (NASH) is a prominent cause of liver-related death that poses a threat to global health and is characterized by severe hepatic steatosis, lobular inflammation, and ballooning degeneration. To date, no Food and Drug Administration-approved medicine is commercially available. The Chaihu Guizhi Ganjiang Decoction (CGGD) shows potential curative effects on regulation of blood lipids and blood glucose, mitigation of organism inflammation, and amelioration of hepatic function. However, the overall regulatory mechanisms underlying its effects on NASH remain unclear. PURPOSE This study aimed to investigate the efficiency of CGGD on methionine- and choline-deficient (MCD)-induced NASH and unravel its underlying mechanisms. METHODS A NASH model of SD rats was established using an MCD diet for 8 weeks, and the efficacy of CGGD was evaluated based on hepatic lipid accumulation, inflammatory response, and fibrosis. The effects of CGGD on the intestinal barrier, metabolic profile, and differentially expressed genes (DEGs) profile were analyzed by integrating gut microbiota, metabolomics, and transcriptome sequencing to elucidate its mechanisms of action. RESULTS In MCD-induced NASH rats, pathological staining demonstrated that CGGD alleviated lipid accumulation, inflammatory cell infiltration, and fibrosis in the hepatic tissue. After CGGD administration, liver index, liver weight, serum alanine aminotransferase (ALT), and aspartate aminotransferase (AST) contents, liver triglycerides (TG), and free fatty acids (FFAs) were decreased, meanwhile, it down-regulated the level of proinflammatory mediators (TNF-α, IL-6, IL-1β, MCP-1), and up-regulated the level of anti-inflammatory factors (IL-4, IL-10), and the expression of liver fibrosis markers TGFβ, Acta2, Col1a1 and Col1a2 were weakened. Mechanistically, CGGD treatment altered the diversity of intestinal flora, as evidenced by the depletion of Allobaculum, Blautia, norank_f_Erysipelotrichaceae, and enrichment of the probiotic genera Roseburia, Lactobacillus, Lachnoclostridium, etc. The colonic histopathological results indicated that the gut barrier damage recovered in the CGGD treatment group, and the expression levels of colonic short-chain fatty acids (SCFAs)-specific receptors FFAR2, FFAR3, and tight junction (TJs) proteins ZO-1, Occludin, Claudin-1 were increased compared with those in the model group. Further metabolomic and transcriptomic analyses suggested that CGGD mitigated the lipotoxicity caused by glycerophospholipid and eicosanoid metabolism disorders by decreasing the levels of PLA2G4A, LPCAT1, COX2, and LOX5. In addition, CGGD could activate the inhibitory lipotoxic transcription factor PPARα, regulate the proteins of FABP1, APOC2, APOA2, and LPL to promote fatty acid catabolism, and suppress the TLR4/MyD88/NFκB pathway to attenuate NASH. CONCLUSION Our study demonstrated that CGGD improved steatosis, inflammation, and fibrosis on NASH through enhancing intestinal barrier integrity and alleviating PPARα mediated lipotoxicity, which makes it an attractive candidate for potential new strategies for NASH prevention and treatment.

ETHNOPHARMACOLOGICAL RELEVANCE Nonalcoholic fatty liver disease (NAFLD) is a manifestation of metabolic syndrome in the liver and the leading cause of chronic liver disease worldwide. Digeda-4 decoction (DGD-4) is a commonly prescribed Mongolian herbal drug for treating acute and chronic liver injury and fatty liver. However, the mechanisms underlying the improvement of dislipidemia and liver injury via treatment with DGD-4 remain unclear. Disassembling a prescription is an effective approach to studying the effects and mechanisms underlying Mongolian medicine prescriptions. By disassembling a prescription, it is feasible to discover effective combinations of individual herbs to optimize a given prescription. Accordingly, we disassembled DGD-4 into two groups: the single Lomatogonium rotatum (L.) Fries ex Nym (LR) (DGD-1) and non-LR (DGD-3). AIM OF THIS STUDY To study whether DGD-4 and its disassembled prescriptions have protective effects against tyloxapol (TY)-induced NAFLD and to explore the underlying mechanisms of action and compatibility of prescriptions. MATERIAL AND METHODS NAFLD mice were developed by TY induction. Biochemical horizontal analyses, enzyme-linked immunosorbent assay, and liver histological staining were performed to explore the protective effects of DGD-4 and its disassembled prescriptions DGD-3 and DGD-1. Furthermore, we performed immunohistochemical analyses and Western blotting to further explore the expression of target proteins. RESULTS DGD-4 and its disassembled prescriptions could inhibit TY-induced dislipidemia and liver injury. In addition, DGD-4 and its disassembled prescriptions increased the levels of p-AMPKα and p-ACC, but decreased the levels of SREBP1c, SCD-1, SREBP-2, and HMGCS1 proteins. The activation of lipid metabolic pathways SIRT1, PGC-1α, and PPARα improved lipid accumulation in the liver. Moreover, DGD-4 could inhibit hepatocyte apoptosis and treat TY-induced liver injury by upregulating the Bcl-2 expression, downregulating the expression of Bax, caspase-3, caspase-8, and the ratio of Bax/Bcl-2, and positively regulating the imbalance of oxidative stress (OxS) markers (such as superoxide dismutase [SOD], catalase [CAT], malondialdehyde [MDA], and myeloperoxidase [MPO]). DGD-1 was superior to DGD-3 in regulating lipid synthesis-related proteins such as SREBP1c, SCD-1, SREBP-2, and HMGCS1. DGD-3 significantly affected the expression of lipid metabolic proteins SIRT1, PGC-1α, PPARα, apoptotic proteins Bcl-2, Bax, caspase-3, caspase-8, and the regulation of Bax/Bcl-2 ratio. However, DGD-1 showed no regulatory effects on Bax and Bcl-2 proteins. CONCLUSION This study demonstrates the protective effects of DGD-4 in the TY-induced NAFLD mice through a mechanism involving improvement of dyslipidemia and apoptosis by regulating the AMPK/SIRT1 pathway. Although the Monarch drug DGD-1 reduces lipid accumulation and DGD-3 inhibits apoptosis and protects the liver from injury, DGD-4 can be more effective overall as a therapy when compared to DGD-1 and DGD-3.

ErChen and YinChen decoction (ECYCD) is an effective traditional Chinese medicine and has been widely used in traditional Chinese medicine to treat nonalcoholic steatohepatitis (NASH), with good curative effects. However, the specific mechanisms underlying these effects are unclear. In this study, we determined the efficacy of ECYCD in a high-fat diet-induced NASH rat model, established by 8-week administration of a high-fat diet. ECYCD was administered daily for 4 weeks, after which the rats were euthanized. The results demonstrated that ECYCD ameliorated high-fat diet-induced NASH, as evidenced by decreased liver indexes, reduced hepatic lipid deposition and liver injury, lower serum biochemistry markers (including low-density lipoprotein), and reduced HOMA-IR scores. Moreover, levels of free fatty acids, tumor necrosis factor, and malondialdehyde were decreased, whereas glutathione was increased in the liver. Serum high-density lipoprotein was also increased in the liver, and ECYCD regulated the c-Jun N-terminal kinase 1 (JNK1) signaling pathway by decreasing the levels of JNK1 protein, JNK1 mRNA, activator protein- (AP-) 1 protein, AP-1 mRNA, and phospho-insulin receptor substrate- (IRS-) 1ser307 and increasing phopsho-PKBser473 levels. These results suggested that ECYCD could ameliorate high-fat diet-induced NASH in rats through JNK1 signaling. ECYCD may be a safe therapeutic option for the treatment of NASH.

ETHNOPHARMACOLOGICAL RELEVANCE Non-alcoholic fatty liver diseases (NAFLD), including nonalcoholic steatohepatitis (NASH) and cirrhosis, are strongly associated with insulin resistance and glucose intolerance. Gan-tang-yi decoction (GTYD), as a famous classical prescription effectively reduced the elevated level of blood glucose, and improved insulin resistance in cirrhotic patients. However, its specific chemical compositions and molecular mechanisms are still unclear. AIM OF THE STUDY The current study aimed to investigate the active ingredients of GTYD and molecular mechanisms underlying the effect of GTYD against insulin resistance of NASH and cirrhosis. MATERIALS AND METHODS Active ingredients of GTYD were evaluated by UPLC-Q-TOF/MS in both positive and negative ion modes. The TCMSP database and SwissTarget Prediction database were utilized to identify the major active components and possible targets of GTYD. The diabetes-related targets were screened in the databases of GenCLiP 3, OMIM, and GeneCards. The intersection of these databases was utilized to identify possible GTYD targets for insulin resistance. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed. Protein-protein interaction (PPI) network and hub target proteins were obtained from the STRING database. The hub targets were validated by RT-qPCR analysis. Furthermore, an insulin-resistant rat model of NASH and cirrhosis was created by subcutaneously injecting CCl4 combined with high fat and high sucrose diet to verify the therapeutic effects and the molecular mechanisms of GTYD in vivo. RESULTS A total of 91 compounds were identified from GTYD by UPLC/Q-TOF-MS/MS. After removing duplicates, there were 230 targets associated with both disease and medicines. Targets of GTYD involve many biological processes, such as the positive regulation of cellular metabolic and cellular lipid metabolic processes. KEGG enrichment showed that PI3K/Akt and AGE/RAGE pathways played a prominent role in the treatment. RT-qPCR from the GTYD-treated rats revealed that TNF-α, IL-6, VEGFA, STAT3, IL-1B, and MAPK3 mRNA levels in the insulin resistance of NASH and cirrhosis rats were down-regulated. Up-regulated IRS2/PI3K/Akt and down-regulated AGE/RAGE and MAPK/NF-κB signal pathways were closely related to the therapeutic effect of GTYD against insulin resistance of NASH and cirrhosis rats by biology methods. CONCLUSIONS GTYD functions as a hepato-protective formula in the insulin resistance of NASH and cirrhosis and exerts its effects via IRS2/PI3K/Akt and AGEs/RAGE signaling pathways.

ETHNOPHARMACOLOGICAL RELEVANCE Gan-Jiang-Ling-Zhu (GJLZ) decoction, a classical Chinese herbal prescription, can be applied for the treatment of metabolic diseases including liver steatosis. Although GJLZ decoction has been widely applied clinically for thousands of years, the mechanism of GJLZ decoction behind treatment of nonalcoholic steatohepatitis (NASH) remains relatively unelucidated. AIM OF THE STUDY To elucidate the efficacy of GJLZ decoction in the treatment of NASH and to investigate its underlying mechanisms from an epigenetic perspective. MATERIALS AND METHODS The quality control of chemical components in GJLZ decoction was conducted. C57BL/6J mice with NASH were induced by feeding them a choline-deficient-high-fat-diet (CDHFD), along with GJLZ decoction intervention for 4 weeks. Then NASH phenotypes including histological steatosis, inflammation, hepatic apoptosis, fibrosis, serum liver enzyme and lipid level were measured. N6-methyladenosine (m6A) and transcriptome sequencing were performed. Levels and functions of methyltransferases and different genes were performed by quantitative polymerase chain reaction, immunofluorescence, gene knockdown, oil red O staining and western blotting. RESULTS GJLZ decoction significantly reduced liver weight, liver index and improved hepatic steatosis, and inflammation, as well as inhibited hepatic apoptosis and fibrosis. Moreover, GJLZ decoction significantly reduced the levels of lactate dehydrogenase, aminotransferase, triglyceride, aspartate aminotransferase, and inhibited levels of interleukin 6 and tumor necrosis factor α. Transcriptome and m6A sequencing revealed the landscape of transcriptome and m6A modification influenced by NASH and the following GJLZ decoction intervention. Eleven differential genes were identified, and GJLZ markedly promoted m6A level of UDP glucuronosyltransferase family 2 member A3 (Ugt2a3), to promote its expression. Additionally, GJLZ significantly promoted methyltransferase 14 (METTL14) expression, whereas METTL14 knockdown aggravated hepatocellular steatosis. Finally, METTL14 knockdown significantly reduced the level of Ugt2a3 by promoting its degradation, whereas, Ugt2a3 overexpression could markedly inhibit hepatocellular steatosis. CONCLUSIONS GJLZ decoction demonstrates potential in alleviating CDHFD-induced NASH by modulating the METTL14-m6A-Ugt2a3 axis, offering a novel therapeutic approach for NASH treatment.

Yinchen Linggui Zhugan decoction (YLZD) is an effective and classical traditional herbal prescription for treating the nonalcoholic fatty liver disease (NAFLD) and has been proven to be effective in the regulation of lipid metabolism disorder and attenuate inflammation for a NAFLD rat model. However, the exact underlying mechanism has not been elucidated. In the current study, a NAFLD rat model was established using a high-fat diet (HFD) for 10 weeks, followed by YLZD treatment with 1.92 g/kg/day for 4 weeks to explore the mechanisms of YLZD. Our results showed that YLZD decreased the hepatic lipid deposition, restored the liver tissue pathological lesions, inhibited the expression of oxidative stress, and decreased the inflammatory cytokines levels. Meanwhile, the genes and proteins expressions of SIRT1/Nrf2 signaling pathway together with downstream factors including HO-1 and NQO1 were elevated in the YLZD treated NAFLD rats. For further elaborating the upstream mechanism, short-chain fatty acids (SCFAs) in serum and feces were measured by liquid chromatograph mass spectrometer and gas chromatograph mass spectrometer, and the differences in gut microbiota of rats in each group were analyzed through high-throughput sequencing of 16S rRNA. The results demonstrated that the contents of butyric acid (BA) and total SCFAs in YLZD-treated NAFLD rats were significantly increased in serum and feces. 16S rRNA sequencing analysis illustrated that YLZD intervention led to a modification of the gut microbiota composition, with a decrease of Oribacterium, Lactobacillus and the ratio of Firmicutes/Bacteroides, as well as the increase in SCFAs-producing bacteria such as Christensenellaceae, Clostridia, Muribaculaceae, and Prevotellaceae. Spearman rank correlation analysis indicated that BA and total SCFAs were negatively co-related with oxidative stress-related factors and inflammatory cytokines, while they were positively co-related with SIRT1/Nrf2 pathway related genes and proteins. Furthermore, in vitro study confirmed that BA effectively reduced oxidative stress by activating SIRT1/Nrf2 signaling pathway in L02 cells. Together, the present data revealed YLZD could ameliorate HFD-induced NAFLD in rats by the modulation of SIRT1/Nrf2 signaling pathway and gut microbiota.

Background: Qinglian Hongqu decoction (QLHQD), a traditional Chinese herbal remedy, shows potential in alleviating metabolic issues related to nonalcoholic fatty liver disease (NAFLD). However, its precise mode of action remains uncertain. Objective: This study aims to evaluate the efficacy and mechanisms of QLHQD in treating NAFLD. Methods: This study utilized a NAFLD mouse model to assess the effects of QLHQD on lipid metabolism, including blood lipids and hepatic steatosis, as well as glucose metabolism, including blood glucose levels, OGTT results, and serum insulin. Network pharmacology, bioinformatics, and molecular docking were used to explore how QLHQD may improve NAFLD treatment. Key proteins involved in these mechanisms were validated via WB and immunohistochemistry. Additionally, the expression of downstream pathway targets was examined to further validate the insulin resistance mechanism by which QLHQD improves NAFLD. Results: Animal studies demonstrated that QLHQD alleviated lipid abnormalities, hepatic steatosis, blood glucose levels, the insulin resistance index, and the OGTT results in NAFLD mice (P < 0.05 or 0.01). Network pharmacology and bioinformatics analyses indicated that the effects of QLHQD on NAFLD might involve bile acid secretion pathways. Subsequent validation through Western blotting, immunohistochemistry, and qPCR demonstrated that QLHQD may influence fat metabolism and insulin sensitivity in NAFLD mice via the FXR/TGR5/GLP-1 signaling pathway. Conclusion: QLHQD significantly alleviates glucose and lipid metabolism disorders in a high-fat diet-induced NAFLD mouse model. Its mechanism of action may involve the activation of the FXR/TGR5/GLP-1 signaling pathway in the gut, which reduces lipid accumulation and insulin resistance.

Background Hepatoprotective effects of Chinese herbal formula Jiangzhi Ligan Decoction (JZLGD) against nonalcoholic fatty liver disease (NAFLD) have been demonstrated, but its mechanism is not clear. The aim of this study was to evaluate the protective effects of against high-fat diet (HFD) induced NAFLD in rat, and to further explore the potential molecular mechanism. Methods SD rats were assigned to five different groups: normal control group, NAFLD model group and JZLGD-treated NAFLD group (3 doses of JZLGD: 2.3, 4.6, 9.2 g/kg of body weight, respectively). All the rats were fed a HFD for 18 weeks except the normal control group(a normal diet). After 12 weeks, rats in JZLGD-treated NAFLD group were administered different doses of JZLGD by oral gavage once daily for another period of 6 weeks, and the rest were given the same dosage of normal saline. After the intervention, blood and liver from each sample were carefully removed and analyzed accordingly. Results We found that JZLGD significantly reduced the liver index, levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum. Furthermore, pathological examinations showed that JZLGD markedly reduced liver lipid droplets and improved liver lipid accumulation, NAFLD activity score and ballooning pathology scoring were also decreased. The detection of cytokines showed that JZLGD could significantly reduced the levels of serum inflammatory factors IL-1β, IL-18, tumor necrosis factor α (TNF-α) and IL-6, protected HFD rats from inflammation. In addition, NAFLD treatment, exhibited significant reduction in serum triglyceride (TG), total cholesterol (TC), low density lipoprotein (LDL) and free fatty acid (FFA) when compared to NAFLD control rats. JZLGD intervention also reduced the level of serum lipopolysaccharide (LPS) and the expression of NOD-like receptor family pyrin domain containing 3 (NLRP3), apoptosis-associated speck-like protein containing a CARD (ASC), caspase-1, caspase-11, GSDMD, GSDMD N-terminus (GSDMD-N), IL-1β, IL-18 in the liver. Conclusion These results demonstrate the hepatoprotective effects of JZLGD in NAFLD mice, the effects may be mediated via downregulation of NLRP3 /caspase-1/GSDMD mediated canonical pyroptosis pathway and LPS /caspase-11/GSDMD mediated non-canonical pyroptosis pathway.

ETHNOPHARMACOLOGICAL RELEVANCE Erchen decoction (ECD) is a traditional Chinese herbal formula widely used in clinical practice for the treatment of nonalcoholic fatty liver disease (NAFLD). However, the molecular mechanisms underlying its therapeutic effects have not been fully clarified. AIM OF THE STUDY To explore the pharmacological efficacy and potential molecular mechanisms of ECD in the treatment of NAFLD. MATERIALS AND METHODS UPLC-Q-E-Orbitrap-MS was used to identify the chemical composition of the ECD and ECD-containing serum (ECDS). Network pharmacology, molecular docking, molecular dynamics simulation, transcriptomics, and metabolomics were integrated to predict the underlying mechanisms. In vivo and in vitro models-including high-fat diet- and glucose-fructose water-fed mice, as well as oleic acid- and palmitic acid-induced HepG2 cells-were used to evaluate the therapeutic effects of ECD. An inhibition strategy was applied to investigate the functions of the validated pathways. RESULTS ECD attenuated NAFLD progression both in vivo and in vitro. Thirty-eight active ingredients were identified in the ECDS. Network pharmacological analysis predicted that PPARG and BCL-2 might be central targets, whereas AMPK signalling and apoptosis are likely the main pathways mediating the anti-NAFLD effects of ECD. Molecular docking and molecular dynamics simulation suggested that multiple ECDS ingredients exhibited favourable binding affinities with AMPK and BCL-2, potentially offering therapeutic effects in NAFLD treatment. RNA sequencing analysis revealed that the differentially expressed genes were enriched primarily in the AMPK pathway. Liver metabolomics analysis revealed that the differentially expressed metabolites were associated mainly with oxidative phosphorylation and AMPK signalling. In experimental validation, ECD treatment increased phosphorylated AMPK levels, thereby promoting PPARα activation and upregulating key enzymes involved in β-oxidation (CPT1A and ACOX1) while downregulating PPARγ and its downstream genes associated with fatty acid synthesis (SREBP-1 and FASN), thus improving hepatic lipid metabolism both in vivo and in vitro. Moreover, the combination of ECD with the AMPK inhibitor compound C counteracted the lipid synthesis-reducing and lipid degradation-promoting effects of ECD both in vivo and in vitro. Additionally, ECD reduced the expression of inflammatory cytokines (TNF-α, IL-6, and IL-1β), suppressed ROS production, increased the mitochondrial membrane potential, and inhibited cell apoptosis by modulating the expression of BCL-2, Bax, cleaved PARP and cleaved caspase-3 both in vivo and in vitro. CONCLUSION ECD attenuates NAFLD progression by targeting AMPK-mediated hepatic lipid metabolism and reducing hepatic apoptosis, providing new insights into the anti-NAFLD mechanism of ECD.

ETHNOPHARMACOLOGICAL RELEVANCE Artemisia argyi (AA), a herbal medicine traditionally used in Asian countries, to treat inflammatory conditions such as eczema, dermatitis, arthritis, allergic asthma and colitis. However, the mechanism of action of this plant with regard to hepatitis and other liver-related diseases is still unclear. AIM This study aimed to investigate the effects of AA ethanol extract on NASH-related fibrosis and gut microbiota in a choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD)-induced mouse model. METHODS Male C57BL/6J mice were fed CDAHFD, with or without AA ethanol extract treatment. Biochemical markers, lipid profiles, hepatic mRNA expression levels of key genes, and the fibrosis area were assessed. In vitro, TGF-β-stimulated human hepatic stellate LX-2 cells and mouse primary hepatic stellate cells (mHSCs) were used to elucidate the effects of AA ethanol extract on fibrosis and steatosis. 16S rRNA sequencing, QIIME2, and PICRUST2 were employed to analyze gut microbial diversity, composition, and functional pathways. RESULTS Treatment with the AA ethanol extract improved plasma and liver lipid profiles, modulated hepatic mRNA expression levels of antioxidant, lipolytic, and fibrosis-related genes, and significantly reduced CDAHFD-induced hepatic fibrosis. Gut microbiota analysis revealed a marked decrease in Acetivibrio ethanolgignens abundance upon treatment with the AA ethanol extract, and its functional pathways were significantly correlated with NASH/fibrosis markers. The AA ethanol extract and its active components (jaceosidin, eupatilin, and chlorogenic acid) inhibited fibrosis-related markers in LX-2 and mHSC. CONCLUSION The AA ethanol extract exerted therapeutic effects on CDAHFD-induced liver disease by modulating NASH/fibrosis-related factors and gut microbiota composition. Notably, AA treatment reduced the abundance of the potentially profibrotic bacterium (A. ethanolgignens). These findings suggest that AA is a promising candidate for treating NASH-induced fibrosis.

Gegen Qilian Decoction (GGQLD) is a well-established classic Chinese medicine prescription in treating nonalcoholic steatohepatitis (NASH). However, the molecular mechanism of GGQLD action on NASH is still not clear. This study aimed to assess the anti-NASH effect of GGQLD, and to explore its molecular mechanisms in vivo and in vitro. In HFD-fed rats, GGQLD decreased significantly serum triglyceride (TG), cholesterol (CHO), total bile acid (TBA), low-density lipoprotein (LDL), free fatty acid (FFA) and lipopolysaccharide (LPS) levels, increased levels of differentially expressed proteins (DEPs) Ahcy, Gpx1, Mat1a, GNMT, and reduced the expression of ALDOB. In RAW264.7 macrophages, GGQLD reduced the expression levels of inflammatory factors TNF-α and IL-6 mRNA, and diminished NASH by increasing differentially expressed genes (DEGs) CBS, Mat1a, Hnf4α and Pparα to reduce oxidative stress or lipid metabolism. The results of DEGs verification also showed that GGQLD up-regulated expressions of Hnf4α, Pparα and Cbs genes. In HepG2 cells, GGQLD decreased IL-6 levels and intracellular TG content, and inhibited FFA-induced expression of toll-like receptor 4 (TLR4). In summary, GGQLD abates NASH associated liver injuries via anti-oxidative stress and anti-inflammatory response involved inhibition of TLR4 signal pathways. These findings provide new insights into the anti-NASH therapy by GGQLD.

Tiaoganquzhi Decoction (TGQZD) is a traditional Chinese herbal formulation demonstrated to be a clinically effective treatment for nonalcoholic fatty liver disease (NAFLD), although details concerning its clinical mechanism are poor. This study aimed to explore the mechanism of TGQZD on improvement of inflammatory damage and dyslipidemia caused by NAFLD through the CGI-58/ROS/NLRP3 inflammasome pathway. In our research, the in vivo protective effects of TGQZD on HFD-induced liver injury in rats and in vitro using lipopolysaccharide (LPS)+palmitate (PA)-stimulated HepG-2 cells model. Histological changes were evaluated by hematoxylin-eosin and Oil Red O staining. Inflammatory cytokines and protein expression were analyzed by ELISA, Real time PCR and western blotting. Liver function, blood lipids, free fatty acids (FFA), and reactive oxygen species (ROS) were determined by biochemical detection. Our results indicated that TGQZD exhibited anti-inflammatory activity, reduced the severity of NAFLD and ameliorated the pathological changes. Further, TGQZD improved liver function and lipid metabolism in NAFLD rats. TGQZD lowered serum aspartate aminotransferase, alanine aminotransferase, triglyceride, and total cholesterol levels. TGQZD suppressed the formulation of FFA and ROS. It also reduced the expression and release of the inflammatory cytokine interleukin-1β by promoting CGI-58 expression and inhibiting the expression of FFA, TNF-α, and the NLRP3 inflammasome induced by ROS. TGQZD exhibited anti-inflammatory effects via the CGI-58, ROS and NLRP3 inflammasome pathway in vivo and in vitro, respectively. Our findings demonstrated that TGQZD is a useful and effective therapeutic agent for treating NAFLD via promotion of CGI-58 to inhibit the expression of ROS-induced NLRP3 inflammasome.

ETHNOPHARMACOLOGICAL RELEVANCE Qing-Zhi-Tiao-Gan-Tang or Qing-Zhi-Tiao-Gan Decoction (QZTGT) is based on the compatibility theory of traditional Chinese medicine (TCM), that is a combination of three classical formulae for the treatment of nonalcoholic fatty liver disease (NAFLD). Its pharmacodynamic material basis is made up of quinones, flavanones, and terpenoids. AIM OF THE STUDY This study aimed to look for a promising recipe for treating nonalcoholic steatohepatitis (NASH), a more advanced form of NAFLD, and to use a transcriptome-based multi-scale network pharmacological platform (TMNP) to find its therapy targets. MATERIALS AND METHODS A classical dietary model of NASH was established using MCD (Methionine- and choline-deficient) diet-fed mice. Liver coefficients like ALT, AST, serum TC, and TG levels were tested following QZTGT administration. A transcriptome-based multi-scale network pharmacological platform (TMNP) was used to further analyze the liver gene expression profile. RESULTS The composition of QZTGT was analyzed by HPLC-Q-TOF/MS, a total of 89 compounds were separated and detected and 31 of them were found in rat plasma. QZTGT improved liver morphology, inflammation and fibrosis in a classical NASH model. Transcriptomic analysis of liver samples from NASH animal model revealed that QZTGT was able to correct gene expression. We used transcriptome-based multi-scale network pharmacological platform (TMNP) to predicted molecular pathways regulated by QZTGT to improve NASH. Further validation indicated that "fatty acid degradation", "bile secretion" and "steroid biosynthesis" pathways were involved in the improvement of NASH phenotype by QZTGT. CONCLUSIONS Using HPLC-Q-TOF/MS, the compound composition of QZTGT, a Traditional Chinese prescription, was separated, analyzed and identified systematically. QZTGT mitigated NASH symptoms in a classical dietary model of NASH. Transcriptomic and network pharmacology analysis predicted the potential QZTGT regulated pathways. These pathways could be used as therapeutic targets for NASH.

Objective Sijunzi, Lizhong, and Fuzilizhong decoction were traditional Chinese classic formulations, which are widely used in clinical treatment, and the underlying mechanism is unclear. In this study, we aim to investigate the molecular mechanisms underlying the protective effects of Sijunzi, Lizhong, and Fuzilizhong on nonalcoholic fatty liver disease (NAFLD). Methods Male Wistar rats were fed a high-fat diet for four weeks to induce NAFLD and were thereafter administered Sijunzi (8 g/kg/d), Lizhong (10 g/kg/d), or Fuzilizhong (10 g/kg/d) by gavage for four weeks. Hepatic damage, lipid accumulation, inflammation, autophagy, and peroxisome proliferator-activated receptor-α signaling were evaluated. Results The high-fat diet-fed rats showed typical symptoms of NAFLD, including elevated levels of hepatic damage indicators, increased hepatic lipid deposition and fibrosis, severe liver inflammation, and prominent autophagy. Upon administration of Sijunzi, Lizhong, and Fuzilizhong, liver health was improved remarkably, along with ameliorated symptoms of NAFLD. In addition, NAFLD-suppressed peroxisome proliferator-activated receptor-α signaling was reactivated after treatment with the three types of decoctions. Conclusions The results collectively signify the effective therapeutic and protective functions of Sijunzi, Lizhong, and Fuzilizhong against NAFLD and demonstrate the potential of Chinese herbal medication in mitigating the symptoms of liver diseases. Novelty of the Work. Traditional Chinese herbal medicine has been used for centuries to treat various diseases, but the molecular mechanisms of individual ingredients have rarely been studied. The novelty of our work lies in elucidating the specific signaling pathways involved in the control of NAFLD using three common Chinese herbal decoctions. We suggest that natural herbal formulations can be effective therapeutic agents to combat against NAFLD.

ETHNOPHARMACOLOGICAL RELEVANCE The Huanglian-Wuzhuyu herb pair (HWHP) is a classic Chinese herbal formula consisting of the root of Coptis chinensis Franch and dried, nearly mature scented fruit of Tetradium ruticarpum (A.Juss.) T.G.Hartley. It is widely utilized to treat gastrointestinal and liver diseases such as diarrhea, dysentery, cholestasis, hepatocellular carcinoma, and nonalcoholic steatohepatitis (NASH). However, the mechanism of HWHP in treating NASH remains poorly understood. AIM OF THE STUDY This study investigated the mechanisms of HWHP in NASH treatment via network pharmacology and validated the results through in vivo experiment using mouse models. MATERIALS AND METHODS The compounds and targets corresponding to the active ingredients of HWHP were obtained from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) database. The genes associated with NASH were obtained from the DisGeNET database. Cytoscape software was employed to construct a "drug-ingredient-target-disease" network. Furthermore, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were employed to analyze the related signaling pathways affected by HWHP. Moreover, AutoDock software was used to assess the potential binding affinity between the key targets of the hub pathway and the bioactive compounds. Subsequently, in vivo experiment was conducted to verify the findings of network pharmacology. RESULTS A total of 41 active compounds and 198 targets of HWHP were screened, of which 51 common targets were related to NASH. GO functional enrichment analysis revealed that HWHP may affect NASH by modulating inflammatory response. KEGG pathway enrichment suggested that the NOD-like receptor (NLR) signaling pathway may play an important role in treating NASH. Molecular docking results demonstrated that most HWHP components were successfully docked to NLRP3 with good binding energy. In vivo experiments revealed that HWHP alleviated liver inflammation, improved liver steatosis, reduced TC, TG, LDL-C, ALT, and AST, decreased mRNA expressions of IL-6, IL-18, and TNF-α in the liver, and lowered the expressions of NLRP3, pro-IL-1β, and ASC protein. Also, immunohistochemical findings presented downregulation of caspase-1 and IL-1β by HWHP. CONCLUSIONS The results disclosed that HWHP ameliorates NASH in mice by reducing inflammation and liver steatosis via inhibition of NLRP3 inflammasome. This study revealed the mechanism of HWHP in treating NASH through experiments.

Obesity and metabolic dysfunction-associated steatotic liver disease (MASLD) are major contributors to the rise in metabolic disorders, particularly in developed countries. Despite the need for effective therapies, natural product-based interventions remain underexplored. This study investigated the therapeutic effects of Endarachne binghamiae, a type of brown algae, hot water extract (EB-WE) in ameliorating obesity and MASLD using high-fat diet (HFD)-induced ICR mice for an acute obesity model (4-week HFD feeding) and C57BL/6 mice for a long-term MASLD model (12-week HFD feeding). EB-WE administration significantly reduced body and organ weights and improved serum lipid markers, such as triglycerides (TG), total cholesterol (T-CHO), HDL (high-density lipoprotein), LDL (low-density lipoprotein), adiponectin, and apolipoprotein A1 (ApoA1). mRNA expression analysis of liver and skeletal muscle tissues revealed that EB-WE upregulated Ampkα and Cpt1 while downregulating Cebpα and Srebp1, suppressing lipogenic signaling. Additionally, EB-WE activated brown adipose tissue through Pgc1α and Ucp1, contributing to fatty liver alleviation. Western blot analysis of liver tissues demonstrated that EB-WE enhanced AMPK phosphorylation and modulated lipid metabolism by upregulating PGC-1α and UCP-1 and downregulating PPAR-γ, C/EBP-α, and FABP4 proteins. It also reduced oxidation markers, such as OxLDL (oxidized low-density lipoprotein) and ApoB (apolipoprotein B), while increasing ApoA1 levels. EB-WE suppressed lipid peroxidation by modulating oxidative stress markers, such as SOD (superoxide dismutase), CAT (catalase), GSH (glutathione), and MDA (malondialdehyde), in liver tissues. Furthermore, EB-WE regulated the glucose regulatory pathway in the liver and muscle by inhibiting the expression of Sirt1, Sirt4, Glut2, and Glut4 while increasing the expression of Nrf2 and Ho1. Tentative liquid chromatography–tandem mass spectrometry (LC-MS/MS) analysis for EB-WE identified bioactive compounds, such as pyropheophorbide A and digiprolactone, which are known to have antioxidant or metabolic regulatory activities. These findings suggest that EB-WE improves obesity and MASLD through regulation of metabolic pathways, glucose homeostasis, and antioxidant activity, making it a promising candidate for natural product-based functional foods and pharmaceuticals targeting metabolic diseases.

Metabolic-associated steatotic liver disease (MASLD), the hepatic manifestation of metabolic syndrome, represents a growing global health concern. The intricate pathogenesis of MASLD, driven by genetic, metabolic, epigenetic, and environmental factors, leads to considerable clinical variability. Dysregulation of hepatic lipid metabolism, particularly cholesterol homeostasis, is a critical factor in the progression of MASLD and its more severe form, metabolic dysfunction-associated steatohepatitis (MASH). This review elucidates the multifaceted roles of cholesterol metabolism in MASLD, focusing on its absorption, transportation, biosynthesis, efflux, and conversion. We highlight recent advancements in understanding these processes and explore the therapeutic potential of natural products such as curcumin, berberine, and resveratrol in modulating cholesterol metabolism. By targeting key molecular pathways, these natural products offer promising strategies for MASLD management. Finally, this review also covers the clinical studies of natural products in MASLD, providing new insights for future research and clinical applications.

Abstract Metabolic dysfunction-associated steatotic liver disease (MASLD) is characterized by progressive mitochondrial dysfunction that disrupts hepatocellular metabolism, redox homeostasis, and inter-organelle communication. Hepatic metabolic zonation, maintained by spatially specialized mitochondrial networks, coordinates β-oxidation, oxidative phosphorylation, and lipid synthesis under physiological conditions. Chronic nutrient excess and insulin resistance disrupt this zonal organization, particularly in pericentral hepatocytes, leading to oxidative imbalance, defective mitochondrial quality control (MQC), and lipid accumulation. Mitochondrial injury is not confined to hepatocytes. The release of mitochondrial DNA (mtDNA), cardiolipin, and other mitochondrial danger-associated molecular patterns activates Kupffer cells and hepatic stellate cells through TLR9- and cGAS-STING-dependent pathways, thereby amplifying inflammatory and fibrogenic responses. Recent studies indicate that selected natural compounds improve mitochondrial function by enhancing AMPK-SIRT1-PGC-1α-dependent biogenesis, promoting PINK1/Parkin-mediated mitophagy, and attenuating mito-DAMP–driven innate immune activation. This review integrates liver metabolism and mitochondrial stress signaling pathways, elucidates the mechanistic framework of liver-mitochondrial interactions in MASLD, and explores pharmacological strategies targeting organelles to restore liver metabolic homeostasis.

BACKGROUND Metabolic dysfunction-associated steatotic liver disease (MASLD), is gradually emerging as one of the most prevalent liver diseases worldwide. Previous research demonstrated the involvement of myeloid differentiation factor 2 (MD2), a co-receptor of TLR4, as a key mediator in MASLD pathogenesis. The current study identifies JM-9 as a novel MD2 inhibitor, and focuses on evaluating its potential therapeutic effects in mitigating MASLD progression. METHODS Drug affinity responsive target stability (DARTS) assay and surface plasmon resonance assay were utilized to evaluate the MD2-targeting specificity of JM-9. In vitro, hepatocytes and macrophages were stimulated with palmitic acid (PA) followed by JM-9 treatment. In vivo, a high-fat diet (HFD)-induced MASLD model was established and subjected to JM-9 administration during the last 2 months. RESULTS JM-9 directly bound the Phe76 residue of MD2 to disrupt the PA-induced MD2/TLR4 complex formation, thus further restoring AMPK phosphorylation and inhibiting NF-κB activation to reducing lipid accumulation and inflammation, respectively. In the HFD-mediated MASLD mouse model, JM-9 alleviated the binding between MD2 and TLR4 in the liver and counteracted hepatic TBK1 and p65 activation and AMPK suppression, thereby mitigating liver inflammation, steatosis, and fibrosis. CONCLUSION JM-9, as a novel MD2 inhibitor, holds potential as a viable treatment option for MASLD by playing a dual role in regulating both lipid metabolism and inflammation response.

Metabolic dysfunction-associated steatotic liver disease (MASLD) has a multifactorial and complex pathogenesis. Notably, the disorder of Bile acid (BA) metabolism and lipid metabolism-induced lipotoxicity are the main risk factors of MASLD. Lupeol, traditional regional medicine from Xinjiang, has a long history of use for its anti-inflammatory, anti-tumor, and immune-modulating properties. Recent research suggests its potential as a therapeutic option for MASLD due to its proposed binding capacity to the nuclear BA receptor, Farnesoid X receptor (FXR), hence could represent a therapeutic option for MASLD. In this study, a natural triterpenoid drug lupeol improved BA metabolism and MASLD in mice through the FXR signaling pathway and the gut-liver axis. Furthermore, lupeol effectively restored gut healthiness and improved intestinal immunity, barrier integrity, and inflammation, as indicated by the reconstructed gut flora. Compared with fenofibrate (Feno), lupeol treatment significantly reduced weight gain, fat deposition, and liver injury, decreased serum total cholesterol (TC) and triglyceride (TG) levels, and alleviated hepatic steatosis and liver inflammation. BA analysis showed that lupeol treatment accelerated BA efflux and decreased uptake of BA by increasing hepatic FXR and bile salt export pump (BSEP) expression. Gut microbiota alterations could be related to enhanced fecal BA excretion in lupeol-treated mice. Therefore, consumption of lupeol may prevent HFD-induced MASLD and BA accumulation, possibly via the FXR signaling pathway and regulating the gut microbiota.

Abstract Metabolic dysfunction-associated steatotic liver disease (MASLD) is a progressive liver disorder with a rising prevalence. It begins with lipid accumulation in hepatocytes and gradually progresses to Metabolic-associated steatohepatitis (MASH), fibrosis, cirrhosis, and potentially hepatocellular carcinoma (HCC). The pathophysiology of MASLD is complex and involves multiple factors, with oxidative stress playing a crucial role. Oxidative stress drives the progression of MASLD by causing cellular damage, inflammatory responses, and fibrosis, making it a key pathogenic mechanism. The Nuclear Factor Erythroid 2-Related Factor 2 / Heme Oxygenase-1 (Nrf2/HO-1) signaling axis provides robust multi-organ protection against a spectrum of endogenous and exogenous insults, particularly oxidative stress. It plays a pivotal role in mediating antioxidant, anti-inflammatory, and anti-apoptotic responses. Many studies indicate that activating the Nrf2/HO-1 signaling pathway can significantly mitigate the progression of MASLD. This article examines the role of the Nrf2/HO-1 signaling pathway in MASLD and highlights natural compounds that protect against MASLD by targeting Nrf2/HO-1 activation. The findings indicate that the Nrf2/HO-1 signaling pathway holds great promise as a therapeutic target for MASLD.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is one of the most prevalent chronic liver diseases in the world, with a current global prevalence estimated to exceed 33.6%. Consequently, the search for safe and effective therapeutic agents for MASLD has gained increasing attention. Nuclear factor erythroid 2-related factor 2 (Nrf2), a well-characterized transcription factor, plays a pivotal role in cellular defense against oxidative stress. Accumulating evidence indicates that activating Nrf2 has emerged as a promising therapeutic strategy for MASLD. Notably, numerous natural compounds have been reported to induce the activation of Nrf2 signaling pathway and thereby ameliorating MASLD progression. These natural Nrf2 activators exert beneficial effects on MASLD by attenuating liver oxidative stress, reducing hepatic steatosis, and alleviating liver inflammation and fibrosis. This review aims to elucidate the critical role of Nrf2 in the development and progression of MASLD, and to explore the therapeutic potential of natural compounds that activate Nrf2 as candidate medicinal agents or dietary supplements for the prevention and treatment of MASLD. By providing updated insights into Nrf2-mediated hepatoprotection, this review not only advances our understanding of Nrf2 as a key therapeutic target for MASLD but also supports the discovery and development of natural Nrf2 activators as novel interventions against MASLD.

Insulin resistance (IR) is a central pathogenic driver of metabolic dysfunction-associated steatotic liver disease (MASLD), yet current therapeutic strategies remain insufficient. Diosgenin (DG), a well-characterized natural compound, has been reported to modulate IR-related mechanisms and, thus, holds significant promise for MASLD treatment. In this study, rodent models fed a high-fat diet and HepG2 cells treated with free fatty acids were employed to simultaneously evaluate systemic metabolic outcomes (including hepatic indices and glucose tolerance) and cellular responses (including lipid accumulation and glucose uptake). The findings demonstrate that DG activates the ADIPOR1/APPL1 signaling pathway by elevating circulating adiponectin levels, thereby enhancing GLUT4-dependent glucose uptake, suppressing key gluconeogenic enzymes (PEPCK and G6 Pase), and inhibiting de novo lipogenesis through SREBP-1 downregulation. These mechanistic insights reveal that DG mitigates IR by augmenting adiponectin signaling, thereby providing a scientific basis for the development of phytotherapeutic strategies targeting MASLD.

Introduction Metabolic dysfunction-associated steatotic liver disease (MASLD) constitutes a major global health burden, with limited therapeutic options currently available. Flavonoids derived from natural plants exhibit promising antioxidant and anti-inflammatory bioactivities, making them potential preventive and therapeutic agents for metabolic liver diseases. Citrus grandis ‘Tomentosa’ (CGT) flavonoids have shown hepatoprotective properties in preliminary preclinical investigations. However, a systematic and critical evaluation of the preclinical evidence supporting their application in MASLD, particularly regarding immune-inflammatory regulatory mechanisms, remains absent. This scoping review aimed to comprehensively map and critically appraise preclinical studies concerning the efficacy and molecular mechanisms of CGT flavonoids in MASLD, with a specific focus on immune-inflammatory signaling modulation. Methods A scoping review methodology was applied to synthesize current preclinical evidence on CGT flavonoids for MASLD. Following standardized literature retrieval and screening procedures, a total of 22 eligible studies were included for qualitative evidence synthesis. The pharmacological effects, mechanistic findings, and methodological limitations of existing studies were systematically summarized and critically assessed. Results Synthetic analysis of the included studies demonstrated that CGT flavonoids effectively ameliorate hepatic steatosis, inflammatory responses, and oxidative stress in experimental MASLD models. The underlying mechanisms are preliminarily associated with the modulation of NF-κB, MAPK, and JAK-STAT signaling pathways, as well as the regulation of immune cell polarization. Nevertheless, most mechanistic evidence remains indirect and inconclusive, largely derived from non-CGT preparations or non-MASLD models, with a lack of direct target validation. The current body of evidence presents critical research gaps, including potential experimental artifacts caused by pan-assay interference, insufficient pharmacokinetic profiling, and inadequate rigorous studies validating CGT-specific regulatory effects on hepatic immune components, such as Kupffer cells and intrahepatic T-cell subsets under MASLD pathological conditions. Discussion The present scoping review confirms the preclinical potential of CGT flavonoids as novel botanical candidates for MASLD treatment, primarily through the suppression of hepatic steatosis, oxidative damage, and immune-inflammatory activation. However, existing evidence is methodologically limited and insufficient to support clinical translation. To address current deficiencies and facilitate translational progress, further rigorous mechanistic validation, standardized CGT flavonoid preparation protocols, and well-designed clinical investigations are urgently required. This work provides a balanced, evidence-based, and critical framework to guide future basic research and translational studies targeting CGT flavonoids for MASLD intervention.

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Metabolic dysfunction-associated steatotic liver disease (MASLD)—formerly known as non-alcoholic fatty liver disease (NAFLD)—is the most common chronic liver disease worldwide. Since there is currently no approved pharmacotherapy for MASLD, there is an urgent unmet need for efficacious therapeutics for this disease. Hepar compositum (HC-24) is a multicomponent medicinal product that consists of 24 natural ingredients. It has been shown to have anti-inflammatory properties in an obesity-associated MASLD mouse model, but its potential to reduce MASLD-associated fibrosis had not been explored before this study. Here, we investigated the hepatic anti-inflammatory and anti-fibrotic potential of HC-24 in a streptozotocin (STZ)- and high-fat diet (HFD)-induced model of MASLD. Mice received a single injection of low-dose STZ at 2 days of age, followed by HFD feeding from 4 to 9 weeks of age. Mice were treated every second day with HC-24 or daily with the positive control telmisartan from 6 to 9 weeks of age. A non-diseased control group was included as a healthy reference. An explorative small-scale pilot study demonstrated that HC-24 improved liver histology, resulting in a lower NAFLD activity score and reduced liver fibrosis. A subsequent full study confirmed these effects and showed that HC-24 reduced hepatic inflammation, specifically reducing T helper cell and neutrophil influx, and decreased hepatic fibrosis (with qualitatively reduced collagen type I and type III immunopositivity) in the absence of an effect on body and liver weight, blood glucose or liver steatosis. These results show that HC-24 has hepatoprotective, anti-inflammatory, and anti-fibrotic properties in an STZ- and HFD-induced model of MASLD/MASH, suggesting that this multicomponent medicine has therapeutic potential for MASLD patients.

The occurrence and development of metabolic dysfunction-associated steatotic liver disease (MASLD) are closely related to intestinal flora imbalance, intestinal barrier damage, and gut-liver axis dysfunction. Due to their multi-target regulatory effects and advantages in intestinal microecological intervention, Chinese herbal monomers have shown promising application prospects in the prevention and treatment of MASLD. However, basic research on their toxicity still lags behind, and issues related to safety and clinical translation urgently need attention. This article systematically reviews the research progress on how flavonoids, triterpenoids, alkaloids, and polysaccharides improve hepatic steatosis, inflammatory responses, and metabolic disorders from a toxicological perspective by reshaping the intestinal microbiota, repairing the intestinal mucosal barrier, regulating short-chain fatty acid and bile acid metabolism, and synergistically acting on signaling pathways such as TLR4/NF-kB, FXR, TGR5, SIRT1, and the NLRP3 inflammasome. Furthermore, by combining methods such as 16S rRNA sequencing, metagenomics, metabolomics, and multi-omics integration, the article analyzes their application value and limitations in toxicological mechanism research, and discusses the translational bottlenecks faced by Chinese herbal monomers in pharmacokinetics, bioavailability, quality standardization, targeted delivery, and toxicological safety. Existing evidence indicates that Chinese herbal monomers have a three-in-one intervention advantage of microecological remodeling-metabolic regulation-inflammation inhibition, but their long-term medication safety, toxic target organs, dose-effect/toxicity relationships, and potential drug interactions still need further clarification. This article aims to provide a systematic reference for the safety evaluation and clinical translational research of Chinese herbal monomers in the prevention and treatment of MASLD.

Chlorogenic acid (CGA) is a natural polyphenol found in coffee, tea, vegetables, and fruits. It exhibits strong antioxidant activity and possesses several other biological properties, including anti-inflammatory effects, antimicrobial activity, and insulin-sensitizing properties. Moreover, it may improve lipid and glucose metabolism. This review summarizes the available information on the therapeutic effect of CGA in metabolic dysfunction-associated steatotic liver disease (MASLD). As the literature search engine, the browsers in the PubMed, Scopus, Web of Science databases, and ClinicalTrials.gov register were used. Animal trials and clinical studies suggest that CGA has promising therapeutic potential in treating MASLD and hepatic steatosis. Its mechanisms of action include antioxidant, anti-inflammatory, and anti-apoptotic effects via the activation of the Nrf2 signaling pathway and the inhibition of the TLR4/NF-κB signaling cascade. Furthermore, the alleviation of liver disease by CGA also involves other important molecules such as AMPK and important physiological processes such as the intestinal barrier and gut microbiota. Nevertheless, the specific target cell and key molecule to which CGA is directed remain unidentified and require further study.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is a widespread and potentially severe liver condition characterized by abnormal fat accumulation in the liver unrelated to alcohol consumption. According to the World Health Organization, MASLD is the most prevalent liver disease globally, it affects approximately 25% of the world’s population, nearly two billion individuals. This staggering prevalence underscores the urgent need for effective and safe therapeutic approaches to address this escalating global health concern. Natural products have emerged as promising candidates for preventing and treating MASLD in recent years because of their diverse bioactive compounds and minimal side effects. Well-known natural products such as curcumin, resveratrol, green tea polyphenols, and silymarin exhibit notable hepatoprotective effects by influencing lipid metabolism, mitigating oxidative stress, and reducing inflammation. Ongoing research highlights the potential of phytochemicals from traditional medicinal plants, such as Phyllanthus and Salvia, in ameliorating liver steatosis and fibrosis. These natural products demonstrate the capacity to impede fibrogenesis by interfering with hepatic stellate cell activation, which is pivotal in liver fibrosis development. Recent studies underscore the significance of natural products in modulating the gut–liver axis, where they restore balance to the gut microbiota and enhance intestinal barrier function, which slows the progression of MASLD. Moreover, advancements in nanotechnology facilitate the targeted delivery of natural product-derived compounds, which enhances their bioavailability and therapeutic efficacy. Harnessing the potency of natural products offers a promising avenue for developing novel, safer therapies for MASLD and addressing a critical global health concern with far-reaching implications for public well-being.

Metabolic dysfunction-associated steatotic liver disease (MASLD), previously known as non-alcoholic fatty liver disease (NAFLD), is a seriously increasing liver disorder affecting nearly 32% of adults globally. Hepatic triglycerides (TG) accumulation is the hallmark of MASLD, which results from dysregulated lipid and fatty acid uptake, increased de novo lipogenesis (DNL), and decreased lipid removal. More recently, selective autophagy of lipid droplets (LDs), termed lipophagy, has emerged to be closely associated with disrupted hepatic lipid homeostasis. Recent studies have indicated that a series of natural products have shown promise as an alternative approach in attenuating MASLD via regulating lipophagy in vivo and in vitro. Therefore, lipophagy could be a new approach for natural products to be used to improve MASLD. This article aims to provide a comprehensive overview on the interrelationship between dysregulated lipid metabolism, lipophagy, and MASLD pathogenesis. In addition, the role of some natural products as lipophagy modulators and their impact on MASLD will be discussed.

Metabolism dysfunction-associated steatotic liver disease affects approximately 30% of the world’s population, yet there is only one approved treatment option applicable to more advanced disease. Many individuals consume natural health products for general health and a variety of medical conditions but none are recommended for metabolism dysfunction-associated steatotic liver disease in current European or American guidelines. Nevertheless, human trials indicate that some of these products may be efficacious for treatment of metabolism dysfunction-associated steatotic liver disease and these are supported by mechanistic studies using animal models. This narrative review aims to highlight recent research in human and animal trials on selected natural health products. So far, neither probiotics nor omega-3 polyunsaturated fatty acids have produced convincing, consistent benefits in human randomized controlled trials although studies in mouse models suggest that they have actions can lead to reduction of hepatic steatosis or other markers, such as liver enzymes. Two of the many polyphenols that have been studied were also reviewed here. Trials with resveratrol in humans have not yielded significant results whereas curcumin, the active ingredient in turmeric, appeared to consistently lower steatosis or liver enzymes. Both compounds reduced steatosis in rodent models of MASLD, involving a variety of mechanisms including anti-oxidant, anti-inflammatory and metabolic effects. More, better-designed and powered human trials are required to provide convincing evidence of efficacy of most natural health products.

No abstract available

Background Livsooth Authentic Herbal Formula (LAH) is a novel Chinese herbal medicine that has been previously shown to prevent non-alcoholic fatty liver disease (NAFLD). However, its efficacy in treating obesity and its underlying mechanisms remain unclear. This study uniquely investigates the therapeutic effects of LAH on high-fat diet (HFD)-induced obese mice, focusing on its multi-targeted regulation of metabolic pathways. This research highlights the potential of a multi-component herbal formula in simultaneously activating the AMPK pathway, regulating lipid metabolism, and enhancing antioxidant defenses. By integrating network pharmacology predictions with proteomics analysis, in vivo, and in vitro experiments, this study provides a comprehensive understanding of LAH's mode of action. Materials and methods Mice were fed a high-fat diet (HFD) for 8 weeks, followed by oral treatment with LAH at doses of 615 mg/kg and 2460 mg/kg for 10 weeks. Each treatment group consisted of 6 mice. Body weight, blood biochemistry, and antioxidant enzyme activities were measured. Network pharmacology and proteomics analyses were conducted to identify mechanisms, and in vitro studies validated molecular pathways. Results This study utilized network pharmacology to investigate the therapeutic effects and mechanisms of LAH on obesity. Through relevant databases, 19 major chemical components and 605 potential targets were identified. KEGG pathway analysis identified the AMPK signaling pathway as a key target of LAH. Animal experiments showed that LAH reduced body weight by 16.55 % compared to HFD-induced mice. In addition to weight reduction, LAH significantly improved serum metabolic parameters. Glucose, triglyceride, and cholesterol levels were significantly reduced, and liver function improved, with ALT decreasing from 142.00 ± 32.63 U/L (HFD) to 63.57 ± 33.16 U/L (H-LAH), and AST from 147.20 ± 12.92 U/L (HFD) to 81.71 ± 31.31 U/L (H-LAH). It also enhanced liver antioxidant enzyme activity and reversed oxidative stress. Proteomics analysis revealed that LAH treatment downregulated the expression of FASN, HMGCR, and SREBP1 while upregulating PRKAA1, PRKAA2, ACACA, SOD1, and GSTP1, which are linked to the AMPK pathway and antioxidant mechanisms. In vitro experiments confirmed that LAH ameliorates hepatic steatosis by activating the AMPK pathway, as evidenced by its regulation of p-ACC, p-AMPK, CPT1A, FAS, and SREBP1 protein expression, identifying it as a critical regulator of obesity and NAFLD. Conclusion LAH reduces the expression of FAS and SREBP1 proteins via the AMPK pathway, while enhancing the activity of antioxidant enzymes. These effects promote weight loss and improve hepatic lipid metabolism, supporting its potential as a therapeutic agent for obesity and related metabolic disorders. This provides a theoretical foundation for using LAH in weight management.

Background The ginseng-containing Shentao Ruangan (STR) formula is clinically used for chronic liver conditions, including hepatic fibrosis. Guided by traditional Chinese medicine (TCM) principles, it targets the “deficiency leading to accumulation (Xu Sun Sheng Ji)" pathology in hepatic fibrosis via the “strengthening healthy qi and dispelling pathogenic factors (Fu Zheng Qu Xie)" mechanism. Aims To elucidate how STR attenuates hepatic fibrosis, focusing on its regulation of the fibrotic liver microenvironment. Methods Single-cell RNA sequencing profiled the fibrotic liver microenvironment; GEO analysis confirmed reactive oxygen species (ROS) as a central fibrotic driver. In vitro/in vivo experiments assessed STR's effects on ROS and fibrosis. Mechanisms were explored via RNA sequencing, network pharmacology, molecular docking, Nrf2 inhibitors and Nrf2-knockout (Nrf2−/−) mice. Results Single-cell sequencing identified hepatocyte-hepatic stellate cell (HSC) crosstalk as a core fibrotic feature, with hepatocellular metabolic dysfunction and elevated ROS promoting fibrosis (GEO-validated). STR alleviated fibrosis and suppressed excessive hepatic ROS in vitro and in vivo. Integrated analyses identified ROS modulation as STR's key anti-fibrotic mechanism, with Nrf2 as the critical target. STR activated hepatocellular Nrf2, alleviating oxidative stress, reducing TGF-β expression and secretion, and thus inhibiting HSC activation and extracellular matrix deposition. Molecular docking, Nrf2 inhibitors and Nrf2−/− mouse studies confirmed Nrf2's essential role. Conclusion This study links hepatocyte-HSC crosstalk/ROS-TGF-β-driven activation in hepatic fibrosis to TCM's “Xu Sun Sheng Ji”, while STR's Nrf2-mediated antioxidant and anti-fibrotic actions embody “Fu Zheng Qu Xie”, integrating TCM with modern medicine. STR, a ginseng-containing formula, is a promising targeted fibrosis therapy, advancing TCM-modern medicine integration.

Metabolic dysfunction-associated steatotic liver disease (MASLD) was once known as non-alcoholic fatty liver disease (NAFLD). MASLD and its progressive form metabolic dysfunction-associated steatohepatitis (MASH) have become significant challenges in global public health with their incidence rates showing a persistent upward trend. MASLD is the result of multiple factors acting simultaneously. Multiple biochemical cycles and molecular pathways are implicated in the pathogenesis of MASLD/MASH, such as the phosphoinositide 3-kinase (PI3K)/ protein kinase B (AKT) signaling pathway. Natural products are considered a treasure for new drug discovery and are of great value to medicine. Natural products mediate their therapeutic effects on MASLD/MASH, at least in part, by modulating the PI3K/AKT signaling pathway. However, at present, there are relatively few systematic reviews and summaries in this field both domestically and internationally. This review systematically examines the molecular mechanisms through which the PI3K/AKT signaling pathway contributes to the pathogenesis of MASLD/MASH. Additionally, it places particular emphasis on the potential of natural compounds targeting this pathway as a preventive strategy against MASLD/MASH. It is evident that the PI3K/AKT signaling pathway plays an important role in the progression of MASLD/MASH. Natural products belonging to various classes, such as phenylpropanoids, flavonoids, terpenoids, and alkaloids, can alleviate MASLD/MASH and the mechanisms involve regulation of the PI3K/AKT signaling pathway. Herbal medicines hold significant research potential in modulating the PI3K/AKT signaling pathway to improve MASLD/MASH. An in-depth understanding of these pharmacological mechanisms will help design precise intervention strategies to effectively interrupt or reverse disease progression.

BACKGROUND The incidence of metabolic dysfunction-associated steatotic liver disease (MASLD) is on the rise, while pharmacotherapeutic options remain limited. Eucalyptus globulus fruit is an herbal medicine with empirical lipid-modulating properties. However, its therapeutic potential against MASLD remains uninvestigated. PURPOSE Assess the anti-MASLD efficacy of E. globulus fruit extract (EgE), identify its major active constituent, and uncover underlying mechanisms. METHODS EgE was evaluated for lipid-lowering activity in free fatty acid (FFA)-stimulated hepatocytes, and assessed for therapeutic effects in two mouse models of MASLD. Phytochemical investigation was conducted to characterize its major constituents. The underlying mechanism was investigated using transcriptomics, drug affinity responsive target stability (DARTS), pathway inhibition/knockdown assays, molecular docking, etc. RESULTS: EgE markedly decreased FFA-elevated lipid content in vitro. Consistently, it substantially ameliorated glucose and lipid metabolic disorders, attenuated hepatic steatosis, and mitigated associated pathological liver injury in vivo. Hepatic transcriptomic profiling revealed marked enrichment of lipid metabolism and endoplasmic reticulum stress (ERS) pathways, and subsequent mechanistic studies further identified EgE as an IRE1α regulator that activates the adaptive IRE1α/XBP1s signaling, thereby restraining pathological ERS. Macrocarpal A (MA) was identified as the primary bioactive component. It binds the h1 pocket of IRE1α to increase its dimeric form, elevate XBP1s levels, and thereby exert lipid-lowering efficacy. CONCLUSION EgE exhibits potent anti-MASLD effects by activating the adaptive IRE1α/XBP1s axis, representing a promising novel therapeutic candidate for MASLD management. Its major constituent MA directly engages the h1 pocket of IRE1α to increase dimeric IRE1α - a previously unrecognized binding mode.

ETHNOPHARMACOLOGICAL RELEVANCE Dandelion (also known as Taraxaci Herba), a plant used in traditional Chinese medicine for both medicinal and dietary purposes, serves as a key ingredient in various liver-protecting herbal formulations. Its primary bioactive compound, taraxasterol (TAR), exhibits antioxidant and anti-inflammatory properties and can inhibit lipid synthesis. Nevertheless, the specific mechanism underlying its role in metabolic dysfunction-associated steatotic liver disease (MASLD) remains unclear. MATERIALS AND METHODS The study integrated ethnopharmacological knowledge with modern techniques. Following in vitro assessment of lipid clearance in mouse hepatocytes, network pharmacology analysis was performed to identify potential targets. The therapeutic effects of TAR were subsequently validated in a high-fat diet-induced MASLD mouse model, with a focus on the FXR signaling pathway. RESULTS TAR effectively reduced hepatic lipid accumulation and inflammation, confirming its ethnopharmacological relevance. Network analysis revealed that TAR's multi-target effects converge on the FXR signaling pathway. In vivo experiments demonstrated that TAR significantly alleviated HFD-induced metabolic dysregulation and liver injury through FXR activation, with efficacy abolished upon FXR knockdown. CONCLUSION TAR ameliorates HFD-induced MASLD by modulating and enhancing the expression of hepatic FXR, thereby normalizing glucolipid metabolism and restraining oxidative and inflammatory damage. These findings position TAR as a promising plant-derived candidate for MASLD therapy and provide mechanistic insight into its hepatoprotective action.

Oxidative stress plays a pivotal role in liver damage, leading to various pathologies, such as hepatic fibrosis and hepatocellular injury. While natural products have garnered attention for their hepatoprotective effects, the potential of Gamisoyo-San (GSS) in mitigating oxidative stress-induced liver injury remains insufficiently studied. This study evaluated the protective effects of GSS against arachidonic acid (AA) and iron-induced cytotoxicity in HepG2 cells. Hepatoprotective effects were assessed through cell viability assays, ROS measurement, mitochondrial membrane potential analysis, and immunoblotting to investigate the involvement of antioxidant signaling pathways, including Erk, Akt, and Nrf2/HO-1. GSS treatment significantly improved cell viability, reduced ROS generation, and ameliorated mitochondrial dysfunction induced by AA + iron-mediated oxidative stress. GSS also activated Erk and Akt pathways, which enhanced the nuclear translocation of Nrf2 and upregulated HO-1 expression. Inhibition of the Erk or Akt pathways impaired the hepatoprotective effects of GSS, demonstrating their essential role in mediating these protective actions. Our findings suggest that GSS protects hepatocytes against oxidative stress-induced damage by modulating the Erk, Akt, and Nrf2/HO-1 pathways. These results highlight GSS as a promising herbal medicine for the pharmaco-acupuncture with therapeutic potential for oxidative stress-related liver diseases.

Objective: This study systematically reviewed the effects of herbal medicine for metabolic dysfunction-associated steatotic liver disease (MASLD).Methods: Fifteen randomized controlled trials involving patients with MASLD or non-alcoholic fatty liver disease (NAFLD) accompanied by cardiometabolic risk factors (CMRFs), published between January 2015 and June 2025, were identified from 10 databases. Meta-analysis focused primarily on clinical biochemical markers.Results: Type 2 diabetes mellitus (T2DM) and body mass index were the only factors of CMRFs that appeared during the search. T2DM studies showed a predominance of phlegm-blood stasis patterns. Herbal interventions targeting phlegm-blood stasis patterns increased in addition to existing traditional dampness-heat therapies. Meta-analysis of the studies reporting phlegm-blood stasis patterns in T2DM demonstrated significant improvements in biochemical markers and total effective rate with herbal medicine in combination with standard care.Conclusion: With the transition from NAFLD to MASLD, the Traditional Korean Medicine pathogenesis has expanded to include phlegm-blood stasis, suggesting broader therapeutic potential. Future research should involve rigorous MASLD-based criteria to improve homogeneity and reduce bias.