目的 本研究利用网络药理学、分子对接技术，探索黄芪生脉散治疗糖尿病的潜在作用机制，并结合体外细胞实验对相关信号通路进行初步验证。方法 查阅多个数据库和结合文献筛选黄芪生脉散的有效成分和对应靶点及糖尿病相关靶点，通过DAVID数据库进行疾病预测以及治疗机制预测，将黄芪生脉散有效成分与成分靶点构建黄芪生脉散的成分-靶点网络。此外，选取成分-靶点网络中度值较大的15个关键靶点和FDR值排名前10的疾病构建黄芪生脉散治疗疾病-靶点网络。再进一步使用AutoDock vina软件对黄芪生脉散的有效成分和糖尿病关键靶点进行分子对接。最后利用HepG2细胞胰岛素抵抗（IR）模型对黄芪生脉散提取物的石油醚部位、乙酸乙酯部位、正丁醇部位进行降血糖活性验证，并通过Western-blotting实验探究其石油醚部位对PI3K-Akt信号通路的影响。结果 共挖掘到黄芪生脉散54个有效成分，411个潜在成分靶点及279个糖尿病相关靶点。此外，预测到黄芪生脉散治疗疾病182种，其中2型糖尿病（T2DM）与靶点关联性最强（degree值=12），由此推测黄芪生脉散可能在治疗糖尿病方面具有显著作用。另外，PPI网络中degree > 20的靶点有AKT1、PIK3CA、PIK3R1和VEGFA等，推测这些靶点可能为黄芪生脉散治疗糖尿病的关键靶点。同时，分子对接结果显示黄芪生脉散8个有效成分（degree值≥15）与糖尿病关键靶点有较强的结合能力，并且KEGG富集分析发现与糖尿病相关的主要调控通路为PI3K-Akt信号通路。体外细胞实验发现，黄芪生脉散提取物可浓度依赖性的增加HepG2细胞胰岛素抵抗模型对葡萄糖的摄取，比阳性药盐酸二甲双胍的药效更强。Western-blotting实验结果表明，黄芪生脉散石油醚部位可显著改善HepG2细胞IR模型Akt、GSK3β蛋白表达水平的异常升高以及p-Akt和p-GSK3β的异常降低（与IR组相比P<0.05）。结论 黄芪生脉散的活性成分可能通过作用于INSR、AKT1、PIK3CA等多个糖尿病相关靶点，调节PI3K-Akt信号通路，降低胰岛素抵抗从而发挥对糖尿病的治疗作用。

Objective In this study, network pharmacology and molecular docking techniques were used to explore the potential mechanism of Huangqi Shengmai Powder in the treatment of diabetes, and the related signal pathways were preliminarily verified by in vitro cell experiments.Methods The effective components, corresponding targets and diabetes related targets of Huangqi Shengmai Powder were screened by consulting multiple databases and combined with literature. the disease and treatment mechanism were predicted by DAVID database, and the component-target network of Huangqi Shengmai Powder was constructed by combining the effective components and component targets of Huangqi Shengmai Powder. In addition, 15 key targets with large values in the component-target network and the diseases with the top 10 FDR values were selected to construct the disease-target network for the treatment of Huangqi Shengmai Powder. Furthermore, AutoDock vina software was used to dock the effective components of Huangqi Shengmai Powder and the key targets of diabetes. Finally, HepG2 cell insulin resistance (IR) model was used to verify the hypoglycemic activity of petroleum ether, ethyl acetate and n-butanol of Huangqi Shengmai Powder extract, and the effect of petroleum ether on PI3K-Akt signal pathway was explored by Western-blotting experiment.Results A total of 54 active components and 411 potential component targets and 279 diabetes related targets were found in Huangqi Shengmai Powder. Furthermore, it is predicted that Huangqi Shengmai Powder can treat 182 kinds of diseases, among which type 2 diabetes mellitus (T2DM) had the strongest correlation with the target (Degree value=12). It is speculated that Huangqi Shengmai Powder may play a significant role in the treatment of diabetes. In addition, the targets of degree value >20 in PPI network were AKT1, PIK3CA, PIK3R1 and VEGFA. It is speculated that these targets may be the key targets of Huangqi Shengmai Powder in the treatment of diabetes. At the same time, the results of molecular docking showed that 8 active components of Huangqi Shengmai Powder (degree ≥15) had strong binding ability to the key targets of diabetes, and KEGG enrichment analysis showed that the main regulatory pathway related to diabetes was PI3K-Akt signal pathway. In vitro cell experiments showed that the extract of Huangqi Shengmai Powder could increase the glucose uptake of HepG2 cells in a concentration-dependent manner, which was stronger than that of metformin hydrochloride. The results of Western-blotting experiment showed that the petroleum ether fraction of Huangqi Shengmai Powder could significantly improve the abnormal increase of Akt and GSK3β protein expression and the abnormal decrease of p-Akt and p-GSK3β in HepG2 cell model IR (P<0.05, compared with IR group).Conclusion The active components of Huangqi Shengmai Powder may play a therapeutic role in diabetes by regulating PI3K-Akt signal pathway and reducing insulin resistance by acting on many diabetes related targets such as INSR, AKT1, PIK3CA and so on.

贵州省科学技术厅基础研究计划项目（黔科合基础[2019]1120号):基于网药理学探究常用活血化瘀中药“扰动”TF及相关靶点的研究，负责人：蒋文雯。