目的 运用网络药理学方法分析葛根汤抗溃疡性结肠炎（UC）的分子机制，并通过分子对接和体内实验验证相关核心预测靶点及葛根汤对UC的保护作用。方法 基于中药系统药理学数据库与分析平台（TCMSP）和Swiss Target Prediction数据库筛选葛根汤的有效成分及靶点，在GeneCards、OMIM等疾病数据库获得UC疾病靶点，取有效成分-疾病的交集靶点做蛋白质-蛋白质相互作用（PPI）网络、基因本体（GO）和京都基因与基因组百科全书（KEGG）富集分析。运用Autodock软件进行分子对接验证并用Pymol软件进行可视化。通过3%硫酸葡聚糖钠盐（DSS）构建的UC小鼠模型探讨葛根汤的抗UC作用及对关键靶点蛋白表达的影响。结果 共筛选出葛根汤有效成分250个，有效成分-疾病靶点取交集得346个靶点，核心靶点为蛋白激酶B1（AKT1）、信号转导与转录激活因子3（STAT3）、原癌基因酪氨酸蛋白激酶Src（SRC）、肿瘤坏死因子（TNF）、白细胞介素（IL）-6等。KEGG通路富集分析发现，葛根汤抗UC涉及多条途径： TNF信号通路、IL-17信号通路、PI3K-Akt信号通路等。分子对接结果表明，槲皮素、芹菜素、木犀草素、山柰酚、柚皮素与关键网络中的排名前10的核心蛋白均具有良好的结合亲和力（结合自由能＜-20.92 kJ·mol^-1）。体内实验表明，与模型组相比，葛根汤能够改善小鼠结肠粪便性状，增加结肠长度、结肠质量指数，减轻肠道黏膜病理损伤，显著下调血清促炎因子肿瘤坏死因子-α（TNF-α）、IL-1β水平（P＜0.01、0.001），上调抗炎因子IL-10水平（P＜0.001），显著降低结肠组织SRC蛋白表达（P＜0.001）。结论 葛根汤能够显著缓解UC，其作用机制可能与抑制结肠组织SRC蛋白介导的炎症级联反应密切相关。

Objective The molecular mechanism of Gegen Decoction against ulcerative colitis (UC) was analyzed by network pharmacology, and the relevant core prediction targets and the protective effect of Gegen Decoction on UC were verified by molecular docking and in vivo experiments. Methods The effective components and targets of Gegen Decoction were screened based on the TCMSP and Swiss Target Prediction databases. UC-related targets were obtained from disease databases GeneCards and OMIM etc. The intersection of effective components and disease targets was used to construct a protein-protein interaction (PPI) network, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed. Subsequently, molecular docking was conducted using Autodock software and visualized with Pymol software. Finally, the anti-UC effect of Gegen Decoction and its impact on the expression of key target proteins were investigated in a UC model induced by 3% Dextran Sulfate Sodium Salt (DSS). Results A total of 250 effective components of Gegen Decoction were screened, and 346 targets were obtained by intersecting the effective components with disease targets. The core therapeutic targets include protein kinase B1 (AKT1), signal transducer and activator of transcription 3 (STAT3), proto-oncogene tyrosine-protein kinase Src (SRC), tumor necrosis factor (TNF) and interleukin-6 (IL6), among others. KEGG pathway enrichment analysis revealed that the anti-UC effects of Gegen Decoction involve several pathways, including the TNF signaling pathway, the IL-17 signaling pathway, the PI3K-Akt signaling pathway, etc. Molecular docking results indicated that quercetin, apigenin, luteolin, kaempferol, and naringenin has good binding affinity (Binding energy < -20.92 kJ·mol^-1) with the top 10 core proteins in the key network. In vivo experiments showed that, compared to the DSS group, Gegen Decoction was able to improve the fecal characteristics, increase colon length and colon mass index, reduce intestinal mucosal damage, significantly downregulate serum pro-inflammatory cytokines, including TNF-α and IL-1β (P < 0.01, 0.001), upregulate the anti-inflammatory cytokine IL-10 (P < 0.001), and significantly reduce the expression of SRC protein in colon tissue (P < 0.001). Conclusion Gegen Decoction has the potential to alleviate UC, and its mechanism of action may be closely related to the inhibition of SRC-mediated inflammatory cascade reaction in colon tissue.

R285.5

河北省自然科学基金中医药联合基金重点项目（H2023209038）；河北省高等学校科学技术研究项目（ZD2022141）；华北理工大学大学生创新创业训练计划项目（X2024109）