PDF(Pubmed)
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.
摘要：
背景：非酒精性脂肪性肝病（NAFLD）是最常见的代谢性疾病，与心血管疾病和糖尿病交织在一起。柴胡疏肝散(CSP)是一种对代谢性疾病有显著治疗作用的中药,比如NAFLD。然而,其药理机制仍有待阐明。方法:采用LC-MS/MS测定CSP的主要化合物。对CSP进行了网络药理学研究。根据口服生物利用度选择其潜在的活性成分，药物相似性指数，和植物化学分析。在获得药物靶标和疾病相关靶标之间的交叉基因后，在Cytoscape中可视化了成分-疾病-靶标网络和蛋白质-蛋白质相互作用分析。使用Metascape数据库进行GO和KEGG富集分析。6周龄雄性C57BL/6小鼠高脂高果糖饮食16周加慢性固定应激2周,体内模型,胃内给予CSP或生理盐水。肝脏组织学,甘油三酯和胆固醇水平，ELISA,和RT-PCR用于评估肝脏炎症和脂肪变性。进行免疫组织化学和蛋白质印迹以评估蛋白质水平。结果：通过网络药理学测定，在CSP中总共鉴定出130个潜在的作用于NAFLD的靶基因。包括肿瘤坏死因子（TNF），白细胞介素-6（IL6），白细胞介素-1β（IL-1β），和过氧化物酶体增殖物激活受体γ（PPARG）。KEGG富集分析显示主要通路参与炎症通路,如TNF和NF-κB信号通路，和代谢相关的途径，比如MAPK,HIF-1,FoxO,和AMPK信号通路。体内实验结果表明，CSP可改善肝细胞脂肪变性模型的肝脏炎症，抑制肝脏脂肪酸的合成。更具体地说,CSP治疗显著抑制肿瘤坏死因子α（TNFα）的表达，伴随着TNF受体1（TNFR1）的减少和TNFR1的配体可用性。结论:通过网络药理学和体内验证相结合,这项研究阐明了CSP对NAFLD的治疗作用，减少肝脏炎症和抑制肝脏脂肪酸合成。更具体地说,CSP的抗炎作用至少部分是通过抑制TNFα/TNFR1信号通路介导的。
