Chaihu Shugan powder alleviates liver inflammation and hepatic steatosis in NAFLD mice: A network pharmacology study and in vivo experimental validation

Abstract

Background: Nonalcoholic fatty liver disease (NAFLD) is the most common metabolic disease and is intertwined with cardiovascular disorders and diabetes. Chaihu Shugan powder (CSP) is a traditional Chinese medicine with a significant therapeutic effect on metabolic diseases, such as NAFLD. However, its pharmacological mechanisms remain to be elucidated.Methods: The main compounds of CSP were measured using LC-MS/MS. A network pharmacology study was conducted on CSP. Its potential active ingredients were selected according to oral bioavailability, drug similarity indices, and phytochemical analysis. After obtaining the intersected genes between drug targets and disease-related targets, the component-disease-target network and protein-protein interaction analysis were visualized in Cytoscape. GO and KEGG enrichment analyses were performed using the Metascape database. Six-week-old male C57BL/6 mice fed a high-fat high-fructose diet for 16 weeks plus chronic immobilization stress for 2 weeks, an in vivo model, were administered CSP or saline intragastrically. Liver histology, triglyceride and cholesterol levels, ELISA, and RT-PCR were used to assess hepatic inflammation and steatosis. Immunohistochemistry and western blotting were performed to assess protein levels.Results: A total of 130 potential target genes in CSP that act on NAFLD were identified through network pharmacology assays, including tumor necrosis factor (TNF), interleukin-6 (IL6), interleukin-1β (IL-1β), and peroxisome proliferator-activated receptor γ (PPARG). KEGG enrichment analysis showed that the main pathways were involved in inflammatory pathways, such as the TNF and NF-κB signaling pathways, and metabolism-related pathways, such as the MAPK, HIF-1, FoxO, and AMPK signaling pathways. The results in vivo showed that CSP ameliorated liver inflammation and inhibited hepatic fatty acid synthesis in the hepatocyte steatosis model. More specifically, CSP therapy significantly inhibited the expression of tumor necrosis factor α (TNFα), accompanied by a decrease in TNF receptor 1 (TNFR1) and the ligand availability of TNFR1.Conclusion: Through the combination of network pharmacology and in vivo validation, this study elucidated the therapeutic effect of CSP on NAFLD, decreasing liver inflammation and inhibiting hepatic fatty acid synthesis. More specifically, the anti-inflammatory action of CSP was at least partially mediated by inhibiting the TNFα/TNFR1 signaling pathway.

Transcriptomic analysis reveals pharmacological mechanisms mediating efficacy of Yangyinghuoxue Decoction in CCl4-induced hepatic fibrosis in rats

“…For example, Qiang Zhang et al found that the active metabolite of Radix Bupleuri, β-sitosterol, reduced lipid accretion in hepatocytes processing free aliphatic acids by activating the PPARγ-UCP-1 pathway, thus exerting anti-hepatic fibrosis effects ( Zhang Y. et al, 2022 ). Lei et al (2022) found that the traditional Chinese medicine formula Chaihu Shugan powder (containing Radix Bupleuri as the main metabolite) ameliorated lipid accretion in the livers of mice with non-alcoholic fatty liver disease, inhibited secretion of inflammatory cytokines in hepatic tissues, and inhibited synthesis of aliphatic acids in the liver. The active metabolite of Salvia miltiorrhiza, cryptotanshinone, activates the AMPK signaling pathway, inhibits fat formation, aliphatic acid oxidation, oxidative stress, and inflammatory reactions, thus ameliorating ethanol-induced liver damage ( Nagappan et al, 2019 ).…”

Mechanisms of ferroptosis in nonalcoholic fatty liver disease and therapeutic effects of traditional Chinese medicine: a review

“…Chaihu Shugan powder (CSP) is a Chinese medicine that is used to dredge the liver and regulate Qi, energize blood, and reduce pain ( 85 ), and it is a general remedy for NAFLD. CSP is composed of Radix Bupleuri, Citrus Reticulata , Chuanxiong Rhizoma, Paeoniae Radix Alba, Aurantii Fructus, Cyperi Rhizoma and Radix Glycyrrhizae.…”

“…Chaihu Shugan powder (CSP) is a Chinese medicine that is used to dredge the liver and regulate Qi, energize blood, and reduce pain (85), and it is a general remedy for NAFLD. CSP is composed of Radix…”

Chaihu Shugan prevents cholesterol gallstone formation by ameliorating the microbiota dysbiosis and metabolic disturbance in mice

“…Chaihu Shugan significantly reduced the rate of CGS recurrence compared with the placebo, with minimal adverse effects. Furthermore, Chaihu Shugan has been used to treat a variety of metabolic gastroenterological and hepatobiliary conditions, including chronic gastritis ( Qin et al, 2013 ), irritable bowel syndrome ( Hang et al, 2022 ), hepatic injury ( Jia et al, 2017 ), liver inflammation and hepatic steatosis ( Lei et al, 2022 ), and non-alcoholic fatty liver disease (NAFLD) ( Wang et al, 2022a ). Scientists have hypothesized that the therapeutic effects of Chaihu Shugan are attributed to its ability to modulate gut microbiota dysbiosis and improve metabolic disorders ( Liang et al, 2018 ; Yang et al, 2022 ).…”