目的 基于网络药理学和实验验证探究五味清浊丸对结肠炎的作用机制。方法 采用TCMSP、TCMID数据库筛选五味清浊丸的主要成分，借助PubChem、SwissTargetPrediction数据库检索对应成分的作用靶点。运用GeneCards、OMIM、DrugBank、TTD检索结肠炎靶点。将交集靶点导入STRING 11.5构建“五味清浊丸–结肠炎–靶点”网络，并运用Cytoscape 3.10.0软件筛选核心靶点。使用DAVID 6.8进行基因本体（GO）和京都基因和基因组百科全书（KEGG）通路富集分析。通过TCMSP、PDB数据库获取关键靶点的3D结构。借助科研者之家分子对接平台进行分子对接，应用MOE2020软件进行可视化。采用3%葡聚糖硫酸钠诱导结肠炎小鼠模型，分别设置对照组、模型组、五味清浊丸（0.4、0.8、1.6 g/kg）组及柳氮磺吡啶组。评估结肠长度及组织病理学变化；ELISA法检测血清炎症因子白细胞介素（IL）-6、肿瘤坏死因子-α（TNF-α）、γ-干扰素（IFN-γ）、IL-10水平；Western blotting检测结肠组织中Src蛋白酪氨酸激酶（SRC）、磷脂酰肌醇3-激酶（PI3K）、蛋白激酶B1（Akt1）蛋白表达。结果 网络药理学分析显示，木犀草素、β-谷甾醇等可能是五味清浊丸的关键活性成分，SRC、Akt1、TNF等为核心靶点，且这些靶点显著富集于PI3K/Akt等炎症相关通路。动物实验进一步表明，五味清浊丸能显著延长模型小鼠结肠，减轻结肠组织病理损伤，并能显著降低促炎因子（IL-6、TNF-α、IFN-γ）水平，提升抗炎因子IL-10水平。五味清浊丸能显著抑制模型组小鼠结肠组织中SRC、PI3K、Akt1的蛋白相对表达量（P＜0.05、0.01）。结论 五味清浊丸能有效缓解结肠炎，其作用机制可能与抑制SRC/PI3K/Akt1信号通路的激活，进而减轻肠道炎症反应有关。

Objective To explore the mechanism of action of Wuli Qingzhuo Pills on colitis based on network pharmacology and experimental validation. Methods The main components of Wuwei Qingzhuo Pills were screened using TCMSP and TCMID databases, and the corresponding target sites of the components were retrieved using PubChem and SwissTargetPrediction databases. The target sites of colitis were retrieved using GeneCards, OMIM, DrugBank, and TTD. The intersection target sites were imported into STRING 11.5 to construct the “Wuwei Qingzhuo Pill-colitis-target” network, and the core target sites were screened using Cytoscape 3.10.0 software. GO and KEGG pathway enrichment analysis was performed using DAVID 6.8. The 3D structures of key target sites were obtained from TCMSP and PDB databases. Molecular docking was performed using the Researcher’s Home Molecular Docking Platform, and visualization was carried out using MOE2020 software. A 3% dextran sulfate sodium-induced colitis mice model was established, with control group, model group, Wuwei Qingzhuo Pills (0.4, 0.8, 1.6 g/kg) groups, and sulfasalazine groups. The colonic length and histopathological changes were evaluated, serum inflammatory factors interleukin IL-6, TNF-α, IFN-γ, and IL-10 levels were detected by ELISA. The expression of SRC, PI3K, and Akt1 in colon tissue was detected by Western blotting. Results Network pharmacology analysis showed that luteolin, β-sitosterol, etc. might be the key active components of WuWEI Qingzhuo Pills, and SRC, Akt1, TNF, etc. were the core targets, and these targets were significantly enriched in inflammation-related pathways such as PI3K/Akt. Animal experiments further demonstrated that Wuwei Qingzhuo Pills could significantly extend the colons of model mice, alleviate colonic tissue pathological damage, and significantly reduce pro-inflammatory factors (IL-6, TNF-α, IFN-γ) levels, and increase anti-inflammatory factor IL-10 levels. Wuwei Qingzhuo Pills could significantly inhibit the relative expression of SRC, PI3K, and Akt1 proteins in the colonic tissue of model group mice (P < 0.05, 0.01). Conclusion Wuwei Qingzhuo Pills can effectively alleviate colitis, and its mechanism of action may be related to inhibiting the activation of SRC/PI3K/Akt1 signaling pathway and reducing intestinal inflammatory responses.

R285;R286.5

内蒙古医学科学院公立医院科研联合基金科技项目（2023GLLH0390，2023GLLH0388）；通辽市科技计划项目（TL2024YF001）