摘要
目的:基于网络药理学探讨温补固肾方(WBGSF)治疗糖尿病肾病(DN)的作用机制。方法:利用中药系统药理学数据库与分析平台(TCMSP)筛选WBGSF有效活性成分及潜在靶点,在GeneCard、OMIM和TTD数据库中寻找DN相关靶点基因,选取WBGSF靶点基因与DN相关靶点基因的共同靶点。通过Cytoscape 3.5.0软件构建“WBGSF活性成分-潜在靶点-DN”网络图。运用STRING数据库构建蛋白质互作(PPI)网络图。对潜在靶点基因进行基因本体(GO)富集分析和京都基因和基因组百科全书(KEGG)通路分析。SD大鼠随机分为对照组(Control)、糖尿病肾病组(DN)和WBGSF低、中、高剂量组。采用链脲佐菌素腹腔注射构建DN模型大鼠。模型构建成功后WBGSF各剂量组大鼠分别给予0.92、1.84、3.68g/kgWBGSF灌胃,每日1次,连续给药12周;Control组和DN组大鼠采用生理盐水灌胃,每日1次,连续给药12周。12周后检测各组大鼠血糖、尿素氮、肌酐、24 h尿蛋白和肾脏组织病理学变化、肾组织AGEs和RAGE含量以及RAGE蛋白表达。结果:数据库筛选得到WBGSF有效活性成分81个,与DN相关潜在作用靶基因148个,KEGG富集通路分析主要包括脂质-动脉粥样硬化信号通路、AGE-RAGE信号通路以及流体剪切应力和动脉粥样硬化通路等。与Control组相比,DN大鼠血糖、尿素氮、肌酐以及24h尿蛋白含量均明显升高(P<0.01),肾组织出现明显的病理学改变,肾组织AGEs和RAGE含量升高,RAGE蛋白表达增加(P<0.01)。与DN组相比,WBGSF各剂量组大鼠血清相关生化指标降低,肾组织损伤得到明显恢复,AGEs和RAGE含量降低、RAGE蛋白表达降低(P<0.05或P<0.01)。结论:WBGSF具有多种活性成分,能通过多靶点、多通路发挥DN肾保护作用;动物实验证实,WBGSF能够改善DN肾损伤,其机制与调控AGE-RAGE信号通路相关。
Objective:To investigate the action mechanism of Wenbu Gushen Formula(WBGSF)in treatment of diabetic nephropathy(DN)based on network pharmacology.Methods:TCMSP database and analysis platform was used to screen the active ingredients and potential targets of WBGSF,and DN-related target genes were searched in GeneCard,OMIM and TTD databases to obtain the common target of WBGSF target genes and DN-related target genes.The‘WBGSF active ingredient-potential target-DN’network map was constructed by Cytoscape 3.5.0 software.The protein interaction(PPI)network map was constructed by using STRING database.GO enrichment analysis and KEGG pathway analysis were performed on potential target genes.SD rats were randomly divided into the control group,the DN group and the WBGSF groups of low-dose,medium-dose and high-dose.DN model rats were established by intraperitoneal injection of Streptozotocin.After successful modeling,rats in the WBGSF groups were given WBGSF at the doses of 0.92 g/kg,1.84 g/kg and 3.68 g/kg,respectively,once a day for 12 weeks.The rats in the control group and the DN group were gavaged with normal saline,once a day for 12 weeks.After 12 weeks of intervention,blood glucose,blood urea nitrogen,creatinine,24 h urine protein,the renal histopathological changes,the contents of AGEs and RAGE in renal tissues,and protein expression of RAGE were detected in each group.Results:The database screened 81 active components of WBGSF,and 148 potential target genes related to DN.The KEGG enrichment pathway analysis mainly included lipid-atherosclerosis signaling pathway,AGE-RAGE signaling pathway,and fluid shear stress and atherosclerotic pathways.Compared with those in the control group,blood glucose,blood urea nitrogen,creatinine and 24 h urine protein were significantly heightened in DN rats(P<0.01),and renal tissue showed obvious pathological changes,the contents of AGEs and RAGE in renal tissue were increased,and the expression of RAGE were increased(P<0.01).Compared with those in the DN group,the serumrelated biochemical indexes were decreased,the renal tissue damage was significantly recovered,the contents of AGEs and RAGE were decreased,the expression of RAGE protein was decreased in WBGSF groups(P<0.05,P<0.01).Conclusion:WBGSF has multiple active components and can exert DN nephroprotection through multiple targets and multiple pathways.Animal experiments confirmed that WBGSF can improve renal injury of DN,and its mechanism is related to the regulation of AGE-RAGE signaling pathway.
作者
禹静
刘续春
李振华
YU Jing;LIU Xuchun;LI Zhenhua(Zhumadian Central Hospital Affiliated to Huanghuai College,Zhumadian 463000,China;Zhengzhou Central Hospital,Zhengzhou 450001,China)
出处
《中医药信息》
2022年第12期27-34,共8页
Information on Traditional Chinese Medicine
基金
河南省中医药科学研究专项课题(2019ZY1018)
河南省医学科技攻关计划项目(2018020767)。
