School of Basic Medicine, Chengdu University of Traditional Chinese Medicine, Chengdu, 611137, China.
Menzies Institute for Medical Research, University of Tasmania, Hobart, Tasmania, TAS7000, Australia.
J Ethnopharmacol. 2022 Oct 5;296:115405. doi: 10.1016/j.jep.2022.115405. Epub 2022 May 27.
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.
This study investigated the mechanisms of HWHP in NASH treatment via network pharmacology and validated the results through in vivo experiment using mouse models.
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.
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.
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.
草药对药黄连-吴茱萸对治疗非酒精性脂肪性肝炎的作用机制：网络药理学研究与体内实验验证
本研究通过网络药理学方法探讨黄连-吴茱萸对药（HWHP）治疗非酒精性脂肪性肝炎（NASH）的作用机制，并通过体内实验验证结果。
从中药系统药理学数据库与分析平台（TCMSP）数据库中获取 HWHP 的活性成分及其对应的化合物和靶点，从 DisGeNET 数据库中获取与 NASH 相关的基因。采用 Cytoscape 软件构建“药物-成分-靶标-疾病”网络。同时，利用基因本体（GO）和京都基因与基因组百科全书（KEGG）分析 HWHP 影响的相关信号通路。此外，采用 AutoDock 软件评估枢纽通路关键靶标与生物活性化合物的潜在结合亲和力。随后，进行体内实验以验证网络药理学的研究结果。
筛选出 HWHP 的 41 种活性化合物和 198 个靶标，其中 51 个共同靶标与 NASH 相关。GO 功能富集分析表明，HWHP 可能通过调节炎症反应影响 NASH。KEGG 通路富集分析提示 NOD 样受体（NLR）信号通路可能在治疗 NASH 中发挥重要作用。分子对接结果表明，HWHP 的大多数成分都能成功与 NLRP3 结合，且具有良好的结合能。体内实验结果显示，HWHP 可减轻肝脏炎症，改善肝脏脂肪变性，降低 TC、TG、LDL-C、ALT 和 AST，降低肝脏中 IL-6、IL-18 和 TNF-α 的 mRNA 表达，并下调 NLRP3、pro-IL-1β 和 ASC 蛋白的表达。此外，免疫组织化学结果表明 HWHP 下调了 caspase-1 和 IL-1β 的表达。
该研究结果表明，HWHP 通过抑制 NLRP3 炎性小体减轻炎症和肝脏脂肪变性，从而改善 NASH 模型小鼠的病情。本研究通过实验揭示了 HWHP 治疗 NASH 的作用机制。
