目的:基于基因芯片数据挖掘和网络药理学研究芪归抵挡汤治疗糖尿病肾病(DKD)分子机制。方法:利用相关数据平台，挖掘芪归抵挡汤治疗DKD的活性成分和作用靶点，并进行GO功能和KEGG通路富集分析。最终，通过体内实验验证芪归抵挡汤调控糖尿病肾病的作用机制。结果:芪归抵挡汤共获得349个潜在靶点，GEO数据库分析获得具有显著性差异基因有841个，将二者取交集共获得45个交集靶点，经复合网络筛选出槲皮素、山柰酚、芦荟大黄素、常春藤皂苷元和β-谷甾醇等重要成分;CASP3、EGF、CXCL8、CCL2及ICAM1等治疗DKD的重要核心靶点。GO功能富集显示，药物反应、炎症反应和对缺氧的反应等在生物过程中富集度较高，质膜、细胞溶质和细胞外空间等在细胞组分中富集度高，蛋白质结合、受体结合和整合素结合等在分子功能中富集度较高;KEGG分析表明该方药效基础主要与AGE-RAGE、甲型流感、卡波西肉瘤相关疱疹病毒感染、EB病毒感染和NF-κB等信号通路有关。体内研究发现，与正常对照组比较，模型对照组小鼠FBG含量明显升高，血清Scr、BUN含量明显增加，肾组织中AGE、RAGE、Caspase-3蛋白表达上调(P＜0。01);与模型对照组比较，芪归抵挡汤3。34 g/kg组可有效改善db/db小鼠血糖和肾功能(P＜0。01)，显著减轻肾脏病理损伤，下调AGEs-RAGE信号通路相关的AGE、RAGE、Caspase-3蛋白的表达(P＜0。01);与白藜芦醇组比较，芪归抵挡汤3。34 g/kg组小鼠治疗6 w、12 w FBG含量，Scr含量明显降低，肾组织AGE、RAGE、Caspase-3蛋白表达明显下调(P＜0。05或P＜0。01)。结论:芪归抵挡汤可有效降低糖尿病肾病小鼠的血糖水平、改善肾脏功能及病理损伤，并可调控AGEs-RAGE信号通路，具有清除糖尿病肾病"代谢记忆"潜能。
Mechanism of Qigui Didang Decoction(芪归抵挡汤)in Regulating Metabolic Memory of Diabetic Kidney Disease Based on GEO Chip Data Mining and Network Pharmacology
Objective:To investigate the molecular mechanism of Qigui Didang Decoction(芪归抵挡汤)in the treatment of diabetic kidney disease(DKD)based on gene chip data mining and network pharmacology.Methods:By using the relevant data platform,the active components and target of Qigui Didang Decoction in the treatment of DKD were explored,and the enrichment analysis of gene ontology(GO)function and Kyoto encyclopedia of genes and genomes(KEGG)pathway was conducted.Finally,in vivo experiments were conducted to verify the mecha-nism of Qigui Didang Decoction in regulating DKD.Results:A total of 349 potential targets were obtained from Qigui Didang Decoction,and 841 significantly different genes were obtained by GEO database analysis.A total of 45 intersection targets were obtained.Quercetin,kaempferol,aloe emodin,ivy saponin,and β-sitosterol were screened as the important components of Qigui Didang Decoction by compound network.CASP3,EGF,CXCL8,CCL2,and ICAM1 were the important core targets of Qigui Didang Decoction in treating DKD.GO functional enrichment shows that drug response,inflammatory response,and anoxic response are highly enriched in biological processes,the plasma mem-brane,cytosol,and extracellular space are highly enriched in cellular components,and protein binding,receptor binding,and integrin binding are highly enriched in molecular functions.KEGG analysis shows that the therapeutic effect of the prescription is mainly related to AGE-RAGE,influenza A,Kaposi's sarcoma-associated herpesvirus infection,Epstein-Barr virus infection,and NF-kappa B signaling pathways.In vivo studies show that compared with the normal control group,the FBG level and the serum contents of Scr and BUN in the model control group are significantly increased,and the expression levels of AGE,RAGE,and Caspase-3 protein in the renal tissue are up-regulated(P＜0.01).Compared with the model control group,Qigui Didang Decoction group can effectively improve the blood glucose and renal function of db/db mice(P＜0.01),significantly reduce renal pathological damage,and down-regulate the expressions of AGE,RAGE,and Caspase-3 pro-teins related to AGEs-RAGE signaling pathway(P＜0.01).Compared with the resveratrol group,the Qigui Didang Decoction group has sig-nificant reductions in FBG levels 6 and 12 weeks after treatment,the Scr content is decreased,while the expressions of AGE,RAGE,and Caspase-3 in renal tissue are significantly down-regulated(P＜0.05,P＜0.01).Conclusion:Qigui Didang Decoction can effectively reduce the blood glucose level,improve the kidney function and pathological damage of DKD mice,and regulate the AGEs-RAGE signaling pathway,which has the potential to clear the metabolic memory of DKD.
Qigui Didang Decoction(芪归抵挡汤)Diabetic kidney diseaseMetabolic memoryNetwork pharmacologyGEO Chip Data
万方数据
