OBJECTIVE:CGA combination consisting of Cordyceps sinensis polysaccharide, gypenosides, and amygdalin, is derived from Fuzheng Huayu capsule (a traditional Chinese medicine approved for liver fibrosis) via previous Uniform Design Experimentation. The effect of CGA on metabolic dysfunction-associated steatohepatitis (MASH) and the potential mechanism based on bile acids (BAs) alternative biosynthesis pathway-farnesoid X receptor (FXR) axis are investigated here. METHODS:Alanine aminotransferase, hepatic triglyceride, malondialdehyde (MDA), BAs, fatty acids oxidation (FAO) activity, mRNA of inflammatory cytokines, α-smooth muscle actin (α-SMA), transforming growth factor-β, collagen type I, and FAO enzymes were detected. Liver sections underwent hematoxylin-eosin, Oil Red O, and Sirius red staining, and immunohistochemistry assay of F4/80 and α-SMA. Protein expression of peroxisome proliferator-activated receptor α (PPAR-α), FXR, BAs biosynthesis enzymes, small heterodimer partner (SHP), bile salt export pump (BSEP), adenosine triphosphate-binding cassette sub-family B member 4 (ABCB4) and nuclear factor-kappa B were detected. KEY FINDINGS:CGA ameliorated MASH, restoring FAO, PPAR-α, and FXR, accompanied by increased chenodeoxycholic acid proportion in the FXR agonist BAs pool, BAs alternative biosynthesis, and the FXR targets including SHP, BSEP, and ABCB4. CONCLUSION:CGA ameliorates MASH, promoting the hepatic BAs alternative biosynthesis pathway to activate FXR-PPARα restoring FAO.

To identify serum diagnostic biomarker for damp-heat (DH) pattern in chronic liver diseases using transcriptomics and metabolomics. Patients with chronic hepatitis B (CHB) or metabolic dysfunction-associated fatty liver disease (MAFLD) were categorized into DH and non-DH pattern groups. Serum RNA profiles were analyzed via RNA-seq/microarray, and metabolites were quantified by ultraperformance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS). Biomarker screening and validation employed discovery (88 cases) and validation (85 cases) cohorts, utilizing Rank-in analysis and Least Absolute Shrinkage and Selection Operator (LASSO) regression for gene selection; reverse transcription-polymerase chain reaction (RT-PCR) and targeted UPLC-MS/MS for expression validation; random forest and receiver operating characteristic (ROC) curve analysis (with area under the curve, AUC) for diagnostic assessment. Compared with the non-DH pattern group, patients with DH pattern showed significantly elevated liver injury markers and reduced apolipoprotein A1 (P<0.05). Integrated transcriptome analysis (Rank-in) identified 315 dysregulated gene sets, primarily enriched in chemokine signaling. LASSO selected 27 genes for RT-PCR validation, confirming 5 differential genes. Metabolomics revealed 25 differential metabolites (discovery cohort), with 7 showing ⩾ 2-fold change; 6 maintained consistent trends in validation. A random forest model combining 4 genes (PTPN22, CTSD, TBX21, STAT4) and 2 metabolites (pyroglutamic acid and glutamic acid) achieved a validation AUC of 0.828 for DH diagnosis. A multi-omics diagnostic model incorporating 4 genes and 2 metabolites demonstrates promising diagnostic potential for DH pattern in patients with chronic liver diseases. (registration No. ChiCTR2000037248)

OBJECTIVE:To evaluate the effects of Sanren Decoction (SRD) on metabolic dysfunction-associated steatotic liver disease (MASLD) induced by high-fat diet (HFD) based on the protective effect of hepatocyte mitochondrial function. METHODS:Thirty-six male C57BL/6 mice were randomly assigned to 4 groups using stratified sampling based on body weight, including control, HFD, low-dose (10.09 g/kg) and high-dose (20.18 g/kg) SRD groups (n=9). MASLD model was induced in mice via a 16-week HFD. Liver histopathology was assessed using haematoxylin and eosin (HE) and Oil red O staining, while hepatic triglycerides (TG), serum alanine aminotransferase (ALT), fasting blood glucose (FBG), and fasting serum insulin levels were measured using commercial kits. Hepaticmetabolic profiling were analyzed and differential metabolite analysis was performed using partial least squares discriminant analysis and Kyoto Encyclopedia of Genes and Genomes pathways. Mitochondrial microstructure was assessed by electron microscopy. The protein expressions of respiratory chain complexes I-V were determined by Western blot analysis, while the activities of complexes I and II were measured using commercial kits. RESULTS:In the HFD-induced MASLD model, 4-week SRD treatment improved hepatic steatosis, inflammation, and hepatocyte ballooning (P<0.05). High-dose SRD treatment significantly reduced hepatic TG, ALT levels, and improved insulin sensitivity (both P<0.05). Low-dose SRD significantly reduced hepatic TG and FBG (P<0.05). Metabolomic analysis showed that differential liver metabolites were enriched in tricarboxylic acid cycle (TAC) pathway in mice of high-dose SRD and HFD groups. Electron microscopy showed that high-dose SRD improved mitochondrial morphology and enhanced adenosine triphosphate production and fatty acid oxidation activity (both P<0.05). Additionally, high-dose SRD significantly decreased the contents of hydrogen peroxide and malondialdehyde in liver tissue and increased the content of superoxide dismutase (both P<0.05). Treatment with high-dose SRD resulted in a significant increase in both protein expression and activity of mitochondrial complex II. Additionally, high-dose SRD enhanced the protein expression of mitochondrial complex I (both P<0.05). CONCLUSION:SRD exhibited hepatoprotective effects in MASLD, improving liver morphology, metabolism, and mitochondrial function, suggesting its potential as a therapeutic strategy for MASLD.

BACKGROUND:During the hepatic histological progression in metabolic dysfunction-associated steatotic liver disease (MASLD), the immunological mechanisms play a the pivotal role, especially when progressing to metabolic dysfunction-associated steatohepatitis (MASH). The discovery of the stimulator of interferon genes (STING) marked a significant advancement in understanding the immune system. METHODS:We searched literature on STING involved in MASLD in PubMed to summarise the role of intrahepatic or extrahepatic STING signal pathways and the potential agonists or inhibitors of STING in MASLD. RESULTS:Besides inflammation and type I interferon response induced by STING activation in the intrahepatic or extrahepatic immune cells, STING activation in hepatocytes leads to protein aggregates and lipid deposition. STING activation in hepatic macrophages inhibits autophagy in hepatocytes and promotes hepatic stellate cells (HSCs) activation. STING activation in HSCs promotes HSC activation and exacerbates liver sinusoidal endothelial cells (LSECs) impairment. However, it was also reported that STING activation in hepatic macrophages promotes lipophagy in hepatocytes and STING activation in HSCs leads to HSC senescence. STING activation in LSEC, inhibits angiogenesis. For extrahepatic tissue, STING signalling participates in the regulation of the intestinal permeability, intestinal microecology and insulin action in adipocytes, which were all involved in the pathogenesis of MASLD. CONCLUSION:There're plenty of STING ligands in MASLD. How STING activation affects the intercellular conversation in MASLD deserves thorough investigation.

OBJECTIVE:Jianpi huoxue decoction (JHD), a Chinese herbal formula, is commonly used for treating alcohol-associated liver disease (ALD). This study aimed to investigate the mechanism by which JHD affects intestinal barrier function in ALD rats. METHODS:The Sprague-Dawley rats were randomly divided into three groups: control group, model group and JHD group. They were pair-fed a modified Lieber-DeCarli liquid diet containing alcohol (model group, n=10; JHD group, n=10) or isocaloric maltose dextrin (control group, n=10) for 6 weeks. After 3 weeks of feeding, the mice in the JHD group were given JHD (10 mL/kg/day) by gavage for 3 weeks, and those in the control and model groups received equal amounts of double-distilled water for the same period of time. Afterwards, all the rats were given lipopolysaccharide (LPS) by gavage and sacrificed 3.5 h later. LPS levels were measured in the portal blood to evaluate gut leakage. Transmission electron microscopy (TEM) was used to observe ultrastructural changes in the intestinal tract. Adherens junction (AJ) and tight junction (TJ) proteins were detected by Western blotting, immunofluorescence or immunohistochemistry. RESULTS:JHD ameliorated Lieber-DeCarli liquid diet-induced hepatic steatosis, inflammation and LPS expression. It improved pathological changes in the liver and alleviated intestinal ultrastructure injury. Moreover, it significantly enhanced the integrity of tight junctions by increasing the expression of zonula occludens-1 (ZO-1) and occludin. It suppressed the activation of myosin light chain (MLC) phosphorylation. CONCLUSION:JHD improves intestinal barrier function and reduces gut leakiness in ALD rats.

Background and Aim: Xuefu Zhuyu decoction (XZD), a traditional Chinese medicinal formula, was firstly recorded in the Qing dynasty of ancient China and previously demonstrated to ameliorate hepatic steatosis. In the present study, the effects of XZD on non-alcoholic fatty liver disease (NAFLD) induced by high-fat diet (HFD) were evaluated in mice and the hepatic transcriptome was detected to disclose the potential mechanisms of XZD. Experimental procedure: The effects of XZD (low- and high-dosage) on NAFLD induced by HFD for 16 weeks were evaluated. Obeticholic acid was used as control drug. Body weight, food intake and index of homeostatic model assessment for insulin resistance (HOMA-IR) were analyzed. Hepatic histology were observed in haematoxylin and eosin stained sections and quantified with NAFLD activity score (NAS). Lipid in hepatocytes was visualized by Oil red staining. Alanine aminotransferase (ALT) and hepatic triglyceride (TG) was measured. The hepatic transcriptom was detected with RNA-sequencing and validated with real-time polymerase chain reaction, western-blotting and hepatic quantitative metabolomics. Results: XZD ameliorated hepatic histology of NAFLD mice, accompanied with decreasing fasting insulin, HOMA-IR, NAS, ALT and hepatic TG. The hepatic transcriptom of NAFLD was significantly reversed by XZD treatment, especially the genes enriched in the pathways of arachidonic acid metabolism, fatty acid degradation, cytokine-cytokine receptor interaction and extracellular matrix (ECM) -receptor interaction. The hepatic quantitative metabolomics analysis confirmed fatty acid degradation as the key targeting pathway of XZD. Conclusions: XZD ameliorated NAFLD induced by HFD, which probably correlated closely to the pathways of fatty acid degradation.

Theaflavins are the characteristic polyphenols in black tea which can be enzymatically synthesized. In this review, the effects and molecular mechanisms of theaflavins on obesity and its comorbidities, including dyslipidemia, insulin resistance, hepatic steatosis, and atherosclerosis, were summarized. Theaflavins ameliorate obesity potentially via reducing food intake, inhibiting pancreatic lipase to reduce lipid absorption, activating the adenosine monophosphate-activated protein kinase (AMPK), and regulating the gut microbiota. As to the comorbidities, theaflavins ameliorate hypercholesterolemia by inhibiting micelle formation to reduce cholesterol absorption. Theaflavins improve insulin sensitivity by increasing the signaling of protein kinase B, eliminating glucose toxicity, and inhibiting inflammation. Theaflavins ameliorate hepatic steatosis via activating AMPK. Theaflavins reduce atherosclerosis by upregulating nuclear factor erythropoietin-2-related factor 2 signaling and inhibiting plasminogen activator inhibitor 1. In randomized controlled trails, black tea extracts containing theaflavins reduced body weight in overweight people and improved glucose tolerance in healthy adults. The amelioration on the hyperlipidemia and the prevention of coronary artery disease by black tea extracts were supported by meta-analysis.

血瘀证理论经历代医家实践和总结逐步系统完善.在现代常见慢性肝病中,血瘀证是重要的中医证型,活血化瘀是重要的中医治法.文章梳理了血瘀证的历史源流及其在常见慢性肝病(包括非酒精性脂肪性肝病、酒精性肝病、慢性病毒性肝炎、肝硬化、肝癌)中的病机认识和证候特点,综述了血瘀证在常见慢性肝病中的客观化及活血化瘀法应用研究进展.

非酒精性脂肪性肝病(NAFLD)属于中医"肝癖""胁痛"及"积聚"范畴,痰湿是其核心致病因素,贯穿NAFLD病理进展的始终.文章梳理了中医痰湿理论的历史沿革,结合研究进展归纳了NAFLD中痰湿的形成与过食肥甘、过劳伤气、肝脾失调、肾精不足、痰湿体质密切相关,综述了NAFLD痰湿证在肝脏影像学、血清脂质和炎症因子、肠道菌群及胆汁酸代谢等方面不同于其他证型的特点,以及针对NAFLD中"痰湿"的祛湿化痰、疏肝健脾、补肾化痰的治法方药与针刺,以期为中医药治疗NAFLD提供科学支撑.

Far-western blotting, derived from the western blot, has been used to detect interactions between proteins in vitro, such as receptor-ligand interactions. The insulin signaling pathway plays a critical role in the regulation of both metabolism and cell growth. The binding of the insulin receptor substrate (IRS) to the insulin receptor is essential for the propagation of downstream signaling after the activation of the insulin receptor by insulin. Here, we describe a step-by-step far-western blotting protocol for determining the binding of IRS to the insulin receptor.

OBJECTIVE:To evaluate the effects of Sanren Tang (SRT, ) on a high-fat diet (HFD)-induced non-alcoholic fatty liver disease (NAFLD) in mice and to investigate the hepatic transcriptome regulated by SRT.METHODS:The primary SRT components were identified using ultra-high-performance liquid chromatography-high-resolution accurate mass spectrometry. The SRT-induced pharmacological effects on HFD-induced NAFLD were evaluated in mice for 16 weeks. Obeticholic acid was used as a control drug. Body weight, food intake, and homeostatic model assessment for insulin resistance (HOMA-IR) index were analysed. Hepatic histological changes were observed in haematoxylin and eosin-stained sections and quantified using the NAFLD activity score (NAS). Serum alanine aminotransferase (ALT) and hepatic triglyceride (TG) levels were measured. Lipids in hepatocytes were visualised by Oil red staining. RNA-sequencing was performed to determine the transcriptome profile of the liver tissue. The differentially expressed genes were validated using real-time polymerase chain reaction and Western blotting.RESULTS:Four principal compounds were identified in the SRT: adenosine, amygdalin, luteoloside, and magnolol. SRT ameliorated hepatic histology and lipid deposition in the NAFLD mice, and decreased HOMA-IR, NAS and ALT, and hepatic TG levels. Hepatic transcriptome analysis revealed 232 HFD-regulated genes that were reversed by SRT simultaneously. Retinol metabolism, cytokine-cytokine receptor interaction, and peroxisome proliferator-activated receptor (PPAR) γ signalling were the top three SRT-regulated pathways in NAFLD.CONCLUSIONS:SRT significantly ameliorated HFD-induced NAFLD, which was correlated with the regulation of genes enriched in the retinol metabolism, cytokine-cytokine receptor interaction, and PPARγ signalling pathways.

Ethnopharmacological relevance: Qushi Huayu Decoction (QHD) is a traditional Chinese medicine formula con-sisting of five herbs, which has been used for non-alcoholic fatty liver disease (NAFLD) treatment in clinic for decades in China and validated in several NAFLD animal models. The hepatic de novo lipogenesis (DNL) is enhanced greatly to contribute to steatosis in NAFLD. The spliced form of X-box binding protein 1 (XBP1s) initiates DNL independently of sterol regulatory element-binding protein (SREBP) and carbohydrate-responsive element-binding protein (ChREBP). Aim of the study: To disclose the mechanism of inhibition on hepatic DNL by QHD and the responsible compounds. Methods: The effects of QHD on hepatic DNL were evaluated in mice induced by high-fructose diet (HFru). The effects of the serum-absorbed compounds of QHD on XBP1s were evaluated in HepG2 cells induced by tunica-mycin. Hepatic histology, triglyceride (TG) and nonesterified fatty acids were observed. Hepatic apolipoprotein B100 and very low-density lipoprotein were measured to reflect lipid out-transport. The mRNA expression of XBP1s and its target genes were detected by real-time polymerase chain reaction. The protein expression of TG synthetases and DNL enzymes, and inositol requirement enzyme 1 alpha (IRE1 alpha), phosphorylated IRE1 alpha and XBP1s were detected in liver tissue and HepG2 cells by western-blot. The binding activity of SREBP1, protein expression of ChREBP and XBP1s were detected in the nuclear extracts of liver tissue. Results: Dynamical observing suggested feeding with HFru for 2 weeks was sufficient to induce hepatic lipo-genesis and XBP1s. QHD ameliorated liver steatosis without enhancing out-transport of lipids, accompanied with more inhibitory effects on DNL enzymes than TG synthetases. QHD inhibits the nuclear XBP1s without affecting ChREBP and SREBP1. In QHD, chlorogenic acid, geniposide and polydatin inhibit lipogenesis initiated by XPB1s. Conclusion: QHD probably decreases hepatic DNL by inhibiting XBP1s independent of SREBP1 and ChREBP. Chlorogenic acid, geniposide and polydatin are the potential responsible compounds.

Inhibition of P300 acetyltransferase activity by specific inhibitor C646 has been shown to improve insulin signaling. However, the underlying molecular mechanism of this improvement remains unclear. In this study, we analyzed P300 levels of obese patients and found that they were significantly increased in liver hepatocytes. In addition, large amounts of P300 appeared in the cytoplasm. Inhibition of P300 acetyltransferase activity by C646 drastically increased tyrosine phosphorylation of the insulin receptor protein substrates (IRS1/2) without affecting the tyrosine phosphorylation of the beta subunit of the insulin receptor (IRβ) in hepatocytes in the absence of insulin. Since IRS1/2 requires membrane translocation and binding to inositol compounds for normal functions, we also examined the role of acetylation on binding to phosphatidylinositol(4,5)P2 and found that IRS1/2 acetylation by P300 reduced this binding. In contrast, we show that inhibition of IRS1/2 acetylation by C646 facilitates IRS1/2 membrane translocation. Intriguingly, we demonstrate that C646 activates IRβ's tyrosine kinase activity and directly promotes IRβ interaction with IRS1/2, leading to the tyrosine phosphorylation of IRS1/2 and subsequent activation of insulin signaling even in the absence of insulin. In conclusion, these data reveal the unique effects of C646 in activating insulin signaling in patients with obesity and diabetes.

目的:观察三仁汤各提取部位对高脂饮食诱导的非酒精性脂肪性肝病(non-alcoholic fatty liver disease,NAFLD)小鼠的作用.方法:使用水提醇沉、石油醚萃取技术获得三仁汤提取部位1,2,3,以三仁汤整方和过氧化物酶体增殖物激活受体α/γ激动剂Saroglitazar作为对照药物.将小鼠分为正常组,模型组,三仁汤提取部位1,2,3组,三仁汤对照组及Saroglitazar对照组,每组9只.正常组予对照饮食,其余各组予高脂饮食,第13周开始灌胃给药,16周末取材.观测小鼠体质量、进食量、空腹血糖、空腹血清胰岛素、胰岛素抵抗指数、血清谷丙转氨酶(alanine aminotransferase,ALT)、肝脏病理、肝脏甘油三酯(tri-glyceride,TG)及肝脏 γ-谷氨酰转移酶(γ-glutamyl transferase,GGT).结果:三仁汤各提取部位均显著降低NAFLD小鼠体质量和空腹血清胰岛素,改善肝脏病理、肝脏TG和GGT.提取部位2、3还可显著降低NAFLD小鼠空腹血糖和胰岛素抵抗指数.提取部位1、2还可显著降低NAFLD小鼠血清ALT.结论:三仁汤各提取部位均不同程度改善高脂饮食诱导的小鼠NAFLD,其中提取部位2的作用最接近整方.

Knockout of the transcription factor X-box binding protein (XBP1) is known to decrease liver glucose production and lipogenesis. However, whether insulin can regulate gluconeogenesis and lipogenesis through XBP1 and how insulin activates the inositol-requiring enzyme-XBP1 ER stress pathway remains unexplored. Here, we report that in the fed state, insulin-activated kinase AKT directly phosphorylates inositol-requiring enzyme 1 at S724, which in turn mediates the splicing of XBP1u mRNA, thus favoring the generation of the spliced form, XBP1s, in the liver of mice. Subsequently, XBP1s stimulate the expression of lipogenic genes and upregulates liver lipogenesis as previously reported. Intriguingly, we find that fasting leads to an increase in XBP1u along with a drastic decrease in XBP1s in the liver of mice, and XBP1u, not XBP1s, significantly increases PKA-stimulated CRE reporter activity in cultured hepatocytes. Furthermore, we demonstrate that overexpression of XBP1u significantly increases cAMP-stimulated expression of rate-limiting gluconeogenic genes, G6pc and Pck1, and glucose production in primary hepatocytes. Reexpression of XBP1u in the liver of mice with XBP1 depletion significantly increases fasting blood glucose levels and gluconeogenic gene expression. These data support an important role of XBP1u in upregulating gluconeogenesis in the fasted state. Taken together, we reveal that insulin signaling via AKT controls the expression of XBP1 isoforms and that XBP1u and XBP1s function in different nutritional states to regulate liver gluconeogenesis and lipogenesis, respectively.

Mitochondria are highly dynamic organelles of bacterial origin in eukaryotic cells. These play a central role in metabolism and adenosine triphosphate (ATP) synthesis and in the production and regulation of reactive oxygen species (ROS). In addition to the generation of energy, mitochondria perform numerous other functions to support key developmental events such as fertilization during reproduction, oocyte maturation, and the development of the embryo. During embryonic and neonatal development, mitochondria may have important effects on metabolic, energetic, and epigenetic regulation, which may have significant short- and long-term effects on embryonic and offspring health. Hence, the environment, epigenome, and early-life regulation are all linked by mitochondrial integrity, communication, and metabolism.

ObjectiveTo investigate the effect of Xuefu Zhuyu decoction on nonalcoholic fatty liver disease (NAFLD) and its material basis. MethodsIn experiment 1 for exploring the effect of Xuefu Zhuyu decoction on mice with NAFLD induced by high-fat diet, 50 healthy male C57BL/6J mice were randomly divided into normal group, model group, high- and low-dose Xuefu Zhuyu decoction groups, and obeticholic acid control group, with 10 mice in each group. The mice in the normal group were given control diet, and those in the other groups were given high-fat diet. Gastric administration was started at week 13, and related samples were collected at the end of week 16. Food intake and body weight were recorded, enzyme-linked immunosorbent assay was used to measure the serum level of fasting insulin, fasting blood glucose was measured, and insulin resistance index was calculated. HE staining and NAFLD activity score (NAS) were used to observe liver histopathology in mice, oil red O staining was used to observe lipid deposition, and triglyceride (TG) level in liver tissue and serum alanine aminotransferase (ALT) level were measured. In experiment 2 for exploring the effect of different extracts of Xuefu Zhuyu decoction on mice with NAFLD induced by high-fat diet, the methods of water decocting, water extraction and alcohol precipitation, and petroleum ether extraction were used to obtain the extracts 1, 2, and 3 of Xuefu Zhuyu decoction, and 54 healthy male C57BL/6J mice were randomly divided into normal group, model group, Xuefu Zhuyu decoction extract 1, 2, and 3 groups, and Xuefu Zhuyu decoction control group, with 9 mice in each group. The mice in the normal group were given control diet, and those in the other groups were given high-fat diet. Gastric administration was started at week 13, and related samples were collected at the end of week 16. Food intake and body weight were recorded, and enzyme-linked immunosorbent assay was used to measure the serum level of fasting insulin, fasting blood glucose was measured, and insulin resistance index was calculated. HE and NAS were used to observe liver histopathology in mice, oil red O staining was used to observe lipid deposition, and the levels of TG and gamma-glutamyl transpeptidase (GGT) in liver tissue and the serum level of ALT were measured. The t-test was used for comparison of normally distributed continuous data between two groups; a one-way analysis of variance was used for comparison between multiple groups, and the least significant difference t-test was used for further comparison between two groups. The Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups and further comparison between two groups. ResultsIn experiment 1, compared with the model group, the high- and low-dose Xuefu Zhuyu decoction groups and the obeticholic acid control group had significant reductions in body weight, insulin resistance index, the distribution of vacuolar lipid droplets in liver tissue, intralobular inflammation, the ballooning degeneration of hepatocytes, NAS score, the level of TG in liver tissue, and the serum level of ALT (all P＜0.05). Compared with obeticholic acid, high- and low-dose Xuefu Zhuyu decoction had a significantly better effect in reducing body weight, insulin resistance index, and total NAS score (all P＜0.05), and low-dose Xuefu Zhuyu decoction had a significantly better effect in improving serum ALT (P＜0.05). In experiment 2, compared with the model group, the Xuefu Zhuyu decoction extract 1, 2, and 3 groups had significant reductions in fasting blood glucose, insulin resistance index, the distribution of lipid droplets in liver tissue, intralobular inflammation lesions, the ballooning degeneration of hepatocytes, total NAS score, and the level of TG in the liver (all P＜0.05). Compared with the model group, the extract 1 group had a significant reduction in body weight (P＜0.05); the extract 2 and 3 groups had a significant reduction in the serum level of ALT (P＜0.05); the extract 2 group had a significant reduction in the level of GGT in liver tissue (P＜0.05). The extract 2 of Xuefu Zhuyu decoction had the closest effect to compound Xuefu Zhuyu decoction. ConclusionXuefu Zhuyu decoction and its extracts can help to achieve varying degrees of improvement in NAFLD induced by high-fat diet in mice, and the extract 2 of Xuefu Zhuyu decoction might be the main material basis for Xuefu Zhuyu decoction.

Gut-derived lipopolysaccharide (LPS) leaking through the dysfunctional intestinal barrier contributes to the onset of non-alcoholic steatohepatitis (NASH) by triggering inflammation in the liver. In the present study, a combination consisting of Atractylodes macrocephala polysaccharide (A), chlorogenic acid (C), and geniposide (G) (together, ACG), was shown to ameliorate NASH in mice and reduce hepatic LPS signaling and endotoxemia without decreasing the abundance of identified Gram-negative bacteria through restoring the intestinal tight junctions. Our data indicated that inhibition of LPS gut leakage by the ACG combination contributed to its amelioration of NASH.

Background: Non-alcoholic fatty liver disease (NAFLD), characterized by the excessive accumulation of hepatic triglycerides (TGs), has become a worldwide chronic liver disease. But efficient therapy keeps unsettled. Our previous works show that geniposide and chlorogenic acid combination (namely the GC combination), two active chemical components combined with a unique ratio (67.16:1), presents beneficial effects on high-fat diet-induced NAFLD rodent models. Notably, microarray highlighted the more than 5-fold down-regulated SCD-1 gene in the GC combination group. SCD-1 is an essential lipogenic protein for monounsaturated fatty acids’ biosynthesis and serves as a key regulatory enzyme in the last stage of hepatic de novo lipogenesis (DNL). Methods: NAFLD mice model was fed with 16 weeks high-fat diet (HFD). The pharmacological effects, primarily on hepatic TG, TC, FFA, and liver enzymes, et al. of the GC combination and two individual components were evaluated. Furthermore, hepatic SCD-1 expression was quantified with qRT-PCR, immunoblotting, and immunohistochemistry. Finally, the lentivirus-mediated over-expressed cell was carried out to confirm the GC combination’s influence on SCD-1. Results: The GC combination could significantly reduce hepatic TG, TC, and FFA in NAFLD rodents. Notably, the GC combination presented synergetic therapeutic effects, compared with two components, on normalizing murine hepatic lipid deposition and disordered liver enzymes (ALT and AST). Meanwhile, the robust SCD-1 induction induced by HFD and FFA in rodents and ALM-12 cells was profoundly blunted, and this potent suppression was recapitulated in lentivirus-mediated SCD-1 over-expressed cells. Conclusion: Taken together, our data prove that the GC combination shows a substantial and synergetic anti-lipogenesis effect in treating NAFLD, and these amelioration effects are highly associated with the potent suppressed hepatic SCD-1 and a blunted DNL process.