基于网络药理学和实验验证的黄芪六一汤药效组分促肾小管上皮细胞自噬抗糖尿病肾病的机制研究

目的 通过网络药理学分析及相关的动物实验探讨黄芪六一汤药效组分（HQD）调控肾小管上皮细胞自噬水平抗糖尿病肾病（DN）的机制。方法 采用 DN 大鼠模型考察 HQD 抗 DN 的作用。利用 SwissTargetPrediction 数据库获得 HQD 主要入血成分的相关靶点，并与 GeneCard、DisGeNET 和 OMIM 数据库中筛选的疾病靶点取交集；利用 STRING 数据库和Cytoscape 构建蛋白质-蛋白质相互作用（PPI）网络获得核心靶点；采用 Metascape 数据库对交集靶点进行基因本体（GO）注释及京都基因与基因组百科全书（KEGG）通路富集分析。利用 DN 大鼠模型对预测结果进行验证。结果 与对照组比较，HQD 能显著降低 DN 模型大鼠血糖（FBG）、血肌酐（Scr）、尿素氮（BUN）、总胆固醇（TC）、三酰甘油（TG）、低密度脂蛋白（LDL）、24 h 尿白蛋白（24 h-U-Alb）值（P＜0.05），抑制肾组织细胞凋亡（P＜0.05）。网络药理学研究显示 HQD 主要通过 STAT3、EGFR、VEGFA、JUN、TNF、AKT1、CASP3、mTOR、IL2 等关键靶点，作用于癌症通路、癌症中的蛋白多糖、PI3K-Akt 信号通路、MAPK 等信号通路；动物验证实验显示 HQD 能下调 PI3K/Akt/mTOR 信号通路相关蛋白的表达，上调肾脏组织自噬相关蛋白 Beclin-1、LC3Ⅱ/Ⅰ的表达（P＜0.05），增加肾小管上皮细胞自噬体及自噬溶酶体数量。结论 HQD 抗 DN 的分子机制可能与抑制 PI3K/Akt/mTOR 信号通路激活，促进肾小管上皮细胞自噬水平，抑制细胞凋亡，减轻肾小管上皮细胞损伤有关。

Investigation on mechanism of active constituents of Huangqi Liuyi Decoction regulating autophagy renal tubular epithelial cells against diabetic nephropathy based on network pharmacology and experimental validation

Objective To investigate the mechanism by which the pharmacological component of Huangqi Liuyi Decocotion (HQD) modulates the level of autophagy in renal tubular epithelial cells against diabetic nephropathy (DN) through network pharmacological analysis and related animal experiments. Methods A rat model of DN was used to investigate the anti-diabetic nephropathy effect of HQD. The SwissTargetPrediction database was used to obtain the relevant targets of HQD''s major blood-entry components and intersected with the disease targets screened in GeneCard, DisGeNET and OMIM databases; the STRING database and Cytoscape were used to construct a protein-protein interaction (PPI) network to obtain the core targets; Metascape database was used to analyze the GO and KEGG enrichment of the intersected targets. The predicted results were then validated using a diabetic nephropathy rat model. Results HQD significantly reduced blood glucose (FBG), blood creatinine (Scr), urea nitrogen (BUN), total cholesterol (TC), triacylglycerol (TG), low-density lipoprotein (LDL), and 24 h urinary albumin (24 h-U-Alb) values in rats with DN (P<0.05), and inhibited apoptosis of renal tissue cells (P<0.05). Network pharmacological studies showed that HQD mainly acted on the cancer pathway, proteoglycans in cancer, PI3K-Akt signaling pathway, MAPK and other signaling pathways through key targets such as STAT3, EGFR, VEGFA, JUN, TNF, AKT1, CASP3, mTOR, IL2, etc. Animal validation experiments showed that HQD could down-regulate PI3K/Akt/mTOR signaling pathway-related proteins, up-regulate the expression of autophagy-related proteins Beclin-1 and LC3II/Ⅰ in renal tissues (P<0.05), and increase the number of autophagosomes and autophagic lysosomes in renal tubular epithelial cells.Conclusion The molecular mechanism of HQD against DN may be related to the inhibition of PI3K/ Akt/mTOR signaling pathway activation, promotion of autophagy level in renal tubular epithelial cells, inhibition of apoptosis, and attenuation of renal tubular epithelial cell injury.