目的 探讨芪参益气滴丸对ApoE基因敲除小鼠动脉粥样硬化中Th17细胞的作用及基于网络药理学的靶点分析。方法 选择芪参益气滴丸有效成分作为目标化学成分，输入其化学结构至MetaDrug预测软件，按照口服利用度、类药性为筛选条件对化合物进行筛选，通过逆向分子对接，进而找到相对应的靶标，构建芪参益气滴丸有效成分—Th17细胞靶标网络。将ApoE^-/-小鼠分为三组，给予高脂喂养和不同剂量芪参益气滴丸干预，8周后比较动脉粥样硬化损伤面积，并检测芪参益气滴丸有效成分—Th17细胞靶标蛋白的表达。结果 芪参益气滴丸有效成分包括黄芪甲苷、三七皂苷R1、丹酚酸B、丹酚酸A、丹参素、原儿茶醛、迷迭香酸、人参皂苷Rg1、人参皂苷Rb1、毛蕊异黄酮、刺芒柄花素7-羟基-4'-甲氧基异黄酮，各成分最终靶点数量分别为114、29、28、25、17、13、29、35、31、14、21。25个与Th17细胞分化相关蛋白被确认为靶点蛋白，多条信号通路参与了芪参益气滴丸干预Th17细胞分化过程。组织病理学分析证实芪参益气滴丸可显著减少动脉粥样硬化损伤面积。蛋白印迹实验验证了芪参益气滴丸可以调节TGF-β信号通路、JAK/STAT信号通路中的关键蛋白。结论 芪参益气滴丸可通过干预TGFβ信号通路和JAK/STAT信号通路、减少Th17细胞分化、抑制动脉粥样硬化。

Objective To explore the effect of Qishen Yiqi pill on Th17 cells in atherosclerosis of AopE gene knockout mice and targets analysis based on network pharmacology.Methods The effective components of Qishen Yiqi pill were selected as the target chemical components, and their chemical structures were input to Metadrug prediction software. The compounds were screened according to the oral utilization and drug like conditions. The corresponding targets were found through reverse molecular docking. The target network of effective components of Qishen Yiqi pill and Th17 cell was constructed. ApoE^-/- mice were divided into three groups, which were fed with high-fat diet and treated with different doses of Qishen Yiqi pill. After 8 weeks, the area of atherosclerotic injury was compared, and the target proteins of effective components of Qishen Yiqi pill and Th17 cell were detected.Results The effective components of Qishen Yiqi pill included Astragaloside IV, Notoginsenoside R1, Salvianolic Acid B, Salvianolic Acid A, Danshensu, Protocatechuic Aldehyde, Rosmarinic Acid, Ginsenoside Rg1, Ginsenoside Rb1, Calycosin and Formononetin. The final target number of each component was 114, 29, 28, 25, 17, 13, 29, 35, 31, 14, 21. Twenty-five proteins related to Th17 cell differentiation were identified as target proteins, and multiple signaling pathways were involved in the process of Th17 cell differentiation. Histopathological analysis confirmed that Qishen Yiqi pill could significantly reduce the area of atherosclerotic injury. Western blotting experiments showed that Qishen Yiqi pill could regulate the key proteins in TGF-β signal pathway and JAK/STAT signal pathway.Conclusion Qishen Yiqi pill could reduce Th17 cell differentiation and inhibit atherosclerosis by intervening TGFβ signal pathway and JAK/STAT signal pathway.

国家自然科学基金委员会地区科学基金项目（81960857）：基于TTC39B促进LXR泛素化修饰探讨芪参益气滴丸对动脉粥样硬化胆固醇逆向转运的作用及机制研究，负责人：彭立；贵州中医药大学大学生创新创业训练计划项目（贵中医大创合字（2020）37号）：基于调节性T细胞抑制炎症-胆固醇摄取通路探讨芪参益气滴丸抑制动脉粥样硬化的机制，负责人：张博承。