基于网络药理学探讨七味白术散预防糖尿病肾病的作用机制

Study on the Mechanism of Qiweibaizhusan in Preventing Diabetic Nephropathy Based on Network Pharmacology

目的 采用网络药理学分析七味白术散(QWBZ)治疗糖尿病肾病(DN)的活性成分、靶点、通路,运用分子对接验证成分与通路相关蛋白的结合能力,通过实验药理学验证通路中相关蛋白的表达量。方法 利用公共数据库筛选活性成分及成分靶点、疾病靶点,构建相互作用网络,并进行生物富集数据分析。Autodock分析活性成分与蛋白的对接能力。通过实验药理学分析QWBZ的降糖作用,采用ELISA法检测小鼠血清中氧化因子丙二醛(MDA)、总超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GPX)的含量,采用Western blot法检测肾脏组织的转化生长因子-β1(TGF-β1)和凋亡信号调节激酶1(ASK1)蛋白表达。结果 网络药理学共筛选到93个化学成分,获得33个核心靶点。QWBZ与葡萄糖稳态负调控、凋亡过程正调控、磷脂酰肌醇3-激酶信号通路正调控、丝裂原活化蛋白激酶(MAPK)级联正调控等生物过程的相关性较好;可能是通过AGE-RAGE、HIF-1、AMPK等信号通路发挥预防糖尿病肾病的作用。实验表明,与空白组相比,模型组小鼠SOD、MDA、GPX含量均呈现显著性差异(P<0.05),与模型组相比,QWBZ组SOD活性升高(P<0.05)、MDA含量降低,GPX含量(P<0.01)、肾脏组织的TGF-β1和ASK1蛋白表达明显降低(P<0.01,P<0.05)。结论 QWBZ可下调高脂饮食和STZ导致的血糖升高,降低机体氧化应激水平,降低肾脏组织的TGFβ1和ASK1蛋白表达,其调节作用与MAPK信号通路有关。

Objective Using network pharmacology methods to predict the active composition,target and mode of action of Qiweibaizhusan(QWBZ) in the treatment of diabetic nephropathy(DN),and using molecular docking to verify the binding ability of the ingredients and pathway-related proteins,and experimental pharmacology to explore the effect of QWBZ in the treatment of DN.Methods Screen active ingredients and disease targets through public databases,construct interaction networks and enrichment analysis.Autodock analyzes the docking ability of active ingredients and proteins.Analyze the hypoglycemic effect of QWBZ on DN mice by experimental pharmacology,ELISA method was used to detect the expression of oxidation factor malondialdehyde(MDA),total superoxide dismutase(SOD) and catalase(CAT) in mouse serum,Western blot was used to detect the protein expression of transforming growth factor-β(TGFβ1)and apoptosis signal-regulated kinase 1(ASK1) in kidney tissue.Results A total of 93 chemical components were screened by network pharmacology,and 33 core targets were obtained.QWBZ has a good correlation with biological processes such as negative regulation of glucose homeostasis,positive regulation of apoptosis process,positive regulation of phosphatidylinositol 3-kinase signaling pathway,and positive regulation of mitogen-activated protein kinase(MAPK)cascade.It plays a role in preventing diabetic nephropathy through AGE-RAGE signaling pathway,regulating HIF-1signaling pathway,AMPK signaling pathway and other signaling pathways.The QWBZ group pharmacy experimental group is formulated,and the content is significantly reduced(P<0.01).The content of SOD,MDA,GPX in the model group was significantly different from the characteristics of the blank group(P<0.05).The SOD content of the QWBZ group was significantly different than that of the model group(P<0.05),the MDA content was lower than that of the model group,and the GPX content was significantly different than the model composition(P<0.01).The expression of TGFβ1 and ASK1 protein in kidney tissue was significantly reduced(P<0.01,P<0.05).Conclusion QWBZ can downregulate high-fat diet and STZ to increase blood sugar and reduce the expression of TGFβ1 and ASK1 proteins in kidney tissue.Its regulatory effect is related to signal pathways such as MAPK.