目的 利用网络药理学技术和生物信息学方法及实验探讨芍药汤治疗溃疡性结肠炎(ulcerative colitis,UC)的活性成分、作用靶点及分子机制。方法 利用Pubchem等数据库筛选芍药汤活性成分并预测靶点,通过NCBI数据库筛选HCC相关疾病靶点,构建PPI网络,进行GO功能富集和KEGG筛选可能涉及的生物过程和信号通路,并用AutoDock Tools进行分子对接验证。将40只雄性C57BL/6J小鼠,按体质量随机分为正常组、模型组、美沙拉嗪组,芍药汤组,除正常组外均予以2.5%右旋葡聚糖硫酸钠(DSS)自由饮水5d诱导UC,连续灌胃给药7d后处死小鼠,ELISA检测结肠组织IL-2、IL-1β、IL-6、TNF-α等细胞因子含量,并对结肠组织样本进行病理切片观察。结果 研究筛选出芍药汤的20个活性成分和945个靶点,其中609个与UC相关。通过PPI网络分析确定了VEGFA、AKT1、PTGS2、STAT3等22个关键靶点。GO分析显示409个富集条目,涉及炎症反应对抗原刺激的负调控、炎症反应的正向调节等。KEGG分析发现136条显著富集通路,包括NF-κB信号通路、Toll样受体信号通路(炎症与免疫相关),VEGF信号通路、ErbB信号通路(细胞增殖与凋亡相关)。分子对接显示活性成分与IL6、TNF、TLR4、IL2、IL1B、PTGS2等靶点蛋白表现出较强亲和力,连接形成稳定构象。ELISA最终检测结果显示,DSS诱导的溃疡性结肠炎(UC)小鼠模型结肠组织中多种炎症细胞因子水平显著升高,其中IL-2、IL-1β、IL-6和TNF-α含量均较正常组出现明显上调(p<0.05)。在药物干预后,美沙拉嗪组和芍药汤组均表现出显著的抗炎效果,能够有效降低上述炎症细胞因子在结肠组织中的表达水平(p<0.05)。值得注意的是,与美沙拉嗪组相比,芍药汤对炎症细胞因子的调节作用更为显著(p<0.05)。结论 芍药汤可以通过多成分、多靶点、多途径治疗UC。实验证明芍药汤可通过调节IL-2、IL-1β、IL-6、TNF-α细胞因子等相关细胞因子水平,调节促炎细胞因子的释放水平减轻UC小鼠结肠炎症,改善UC小鼠结肠组织损伤。

Objective To explore the active components, therapeutic targets, and molecular mechanisms of Shaoyao Decoction in the treatment of ulcerative colitis (UC) using network pharmacology techniques, bioinformatics methods, and experimental approaches. Methods Screening active components of Shaoyao Decoction and predicting their targets using databases such as PubChem, screening HCC-related disease targets through the NCBI database, constructing a PPI network, conducting GO functional enrichment and KEGG pathway analysis to identify potential biological processes and signaling pathways involved, and validating with molecular docking using AutoDock Tools. Forty male C57BL/6J mice were randomly divided into normal, model, mesalazine, and Shaoyao Tang groups based on body weight. Except for the normal group, all other groups were induced with ulcerative colitis (UC) by providing 2.5% dextran sulfate sodium (DSS) in drinking water for 5 days. After continuous intragastric administration for 7 days, the mice were sacrificed. The levels of cytokines such as IL-2, IL-1β, IL-6, and TNF-α in colon tissues were measured by ELISA, and pathological sections of colon tissue samples were observed. Results The study identified 20 active components and 945 targets of Shaoyao Tang, among which 609 were related to UC. Through PPI network analysis, 22 key targets including VEGFA, AKT1, PTGS2, and STAT3 were determined. GO analysis revealed 409 enriched terms, involving negative regulation of inflammatory response to antigenic stimulus, positive regulation of inflammatory response, etc. KEGG analysis discovered 136 significantly enriched pathways, including the NF-κB signaling pathway, Toll-like receptor signaling pathway (related to inflammation and immunity), VEGF signaling pathway, and ErbB signaling pathway (related to cell proliferation and apoptosis). Molecular docking revealed that the active ingredients exhibited strong affinity with target proteins such as IL6, TNF, TLR4, IL2, IL1B, and PTGS2, forming stable conformations. The final ELISA results demonstrated that the levels of multiple inflammatory cytokines in the colon tissue of DSS-induced ulcerative colitis (UC) mouse models were significantly elevated, with notable upregulation of IL-2, IL-1β, IL-6, and TNF-α compared to the normal group (p<0.05). Following drug intervention, both the Mesalazine group and the Shaoyao Decoction group exhibited significant anti-inflammatory effects, effectively reducing the expression levels of the aforementioned inflammatory cytokines in the colon tissue (p<0.05). Notably, compared to the Mesalazine group, Shaoyao Decoction demonstrated a more pronounced regulatory effect on inflammatory cytokines (p<0.05).Conclusion Shaoyao Decoction can treat UC through multiple components, multiple targets, and multiple pathways. Experiments have demonstrated that Shaoyao Decoction can alleviate colonic inflammation in UC mice by regulating the levels of related cytokines such as IL-2, IL-1β, IL-6, and TNF-α, and modulating the release levels of pro-inflammatory cytokines, thereby improving colonic tissue damage in UC mice.

(No.82074328)：基于PI3K/AKT通路调控细胞周期和自噬探讨黄芩汤干预结肠"炎﹣癌"转化进程的机制研究,负责人：杨伟鹏。