【摘要】 目的：通过网络药理学、分子对接技术和体外细胞实验，探索和验证白芍对化学性肝损伤的潜在保护作用靶点。方法：首先采用网络药理学方法收集白芍和化学性肝损伤的共同潜在靶点，其次使用分子对接技术拟合化学成分和靶点，随后运用体外细胞实验进行验证。以10 ng/mL TGF-β1激活HSC-LX2，建立肝纤维化细胞模型。用不同浓度的白芍总苷（白芍的活性物质）处理细胞，然后用CCK-8测定其活力，ELISA法测定HSP-90α、PTGS2、IL-6、IL-1β和TNF-α的表达水平，western blot法检测COL-I、COL-III、PPAR-γ和CASP3的蛋白表达水平。结果：通过网络药理学发现白芍有13个关键化合物，保护化学性肝损伤的潜在靶点有IL-6、AKT1、JUN、HSP90AA1、PPARG、PTGS2和CASP3等。GO富集发现涉及对药物的反应、膜筏和肽结合等；KEGG富集发现主要途径涉及脂质与动脉粥样硬化、化学致癌-受体激活等。分子对接显示活性成分芍药苷和芍药内酯苷对HSP90AA1、PTGS2、PPARG和CASP3表现出较强的亲和力。细胞实验验证表明，不同浓度的白芍总苷可抑制活化的HSC-LX2细胞中COL-Ⅰ、COL-Ⅲ、IL-6、TNF-α、IL-1β、HSP-90α和PTGS2的表达，同时增加PPAR-γ和CASP3的表达。结论：白芍对化学性肝损伤的保护作用涉及多组分、多靶点、多途径，主要通过调节HSP90AA1、PTGS2、PPARG、CASP3等靶点发挥作用。白芍总苷是白芍的主要活性化合物，通过减少炎症反应、激活凋亡蛋白和促进活化的HSCs凋亡来保护化学性肝损伤。今后，可以从调节炎症反应和细胞凋亡两个角度研究白芍抗化学性肝损伤的作用机制。更多还原

【Abstract】 Objective: To explore and validate the potential targets of Paeoniae Radix Alba(P. Radix, Bai Shao) in protecting against chemical liver injury through network pharmacology, molecular docking technology,and in vitro cell experiments.Methods: Network pharmacology was used to identify the common potential targets of P. Radix and chemical liver injury. Molecular docking was used to fit the components, which were subsequently verified in vitro. A cell model of hepatic fibrosis was established by activating hepatic stellate cell(HSC)-LX2 cells with 10 ng/m L transforming growth factor-β1. The cells were exposed to different concentrations of total glucosides of paeony(TGP), the active substance of P. Radix, and then evaluated using the cell counting kit-8 assay, enzyme-linked immunosorbent assay, and western blot.Results: Analysis through network pharmacology revealed 13 key compounds of P. Radix, and the potential targets for preventing chemical liver injury were IL-6, AKT serine/threonine kinase 1, jun protooncogene, heat shock protein 90 alpha family class A member 1(HSP90AA1), peroxisome proliferator activated receptor gamma(PPARG), PTGS2, and CASP3. Gene Ontology(GO) enrichment analysis indicated the involvement of response to drugs, membrane rafts, and peptide binding. Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analysis revealed that the main pathways involved lipid and atherosclerosis and chemical carcinogenesis-receptor activation. Paeoniflorin and albiflorin exhibited strong affinity for HSP90AA1, PTGS2, PPARG, and CASP3. Different concentrations of TGP can inhibit the expression of COL-Ⅰ, COL-Ⅲ, IL-6, TNF-a, IL-1β, HSP-90a, and PTGS2 while increasing the expression of PPAR-γ and CASP3 in activated HSC-LX2 cells.Conclusion: P. Radix primarily can regulate targets such as HSP90AA1, PTGS2, PPARG, CASP3. TGP, the main active compound of P. Radix, protects against chemical liver injury by reducing the inflammatory response, activating apoptotic proteins, and promoting the apoptosis of activated HSCs.更多还原