肾舒复健汤对慢性肾衰竭大鼠PTEN/PI3K/AKT信号通路的影响  

Effect of Shenshu Fujian Decoction on PTEN/PI3K/AKT Signaling Pathway in Rats with Chronic Renal Failure

作　　者：罗洪玉 黄学宽[1] 田珈瑜 左玲 沈清[2] 徐珂 王慕南 陈欣 王建伟[1] LUO Hongyu;HUANG Xuekuan;TIAN Jiayu;ZUO Ling;SHEN Qing;XU Ke;WANG Munan;CHEN Xin;WANG Jianwei(School of Traditional Chinese Medicine,Chongqing Medical University,Chongqing 400016,China;Department of Renal Medicine,The First Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]重庆医科大学中医药学院,重庆400016 [2]重庆医科大学附属第一医院肾内科,重庆400016

出　　处：《中药新药与临床药理》2022年第9期1170-1179,共10页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：重庆市卫生和计划生育委员会、重庆市科学技术委员会联合资助项目(ZY201802149);重庆市技术创新与应用发展专项面上项目(cstc2019jscx-msxmX0119)。

摘　　要：目的 基于网络药理学结合动物体内实验探讨肾舒复健汤(黄芪、白术、生地黄等)治疗慢性肾衰竭(CRF)的作用机制。方法 (1)通过TCMSP数据库检索并筛选肾舒复健汤的活性成分,通过Swiss数据库筛选其潜在作用靶点;通过DisGeNET、GeneCards数据库检索CRF疾病相关靶点;通过Venn平台对活性成分潜在作用靶点与CRF疾病相关靶点取交集,获得肾舒复健汤治疗CRF的关键靶点(共同靶点);采用Cytoscape3.7.2软件构建肾舒复健汤治疗CRF的“活性成分-关键靶点”网络,分析核心活性成分;将共同靶点导入STRING数据库,构建蛋白互作(PPI)网络,筛选核心靶点;将共同靶点导入DAVID数据库进行GO功能及KEGG通路富集分析。(2)将大鼠随机分为正常组、模型组、尿毒清组(5 g·kg^(-1))、肾舒复健汤组(11 g·kg^(-1)),每组8只;采用饲喂0.5%腺嘌呤饲料以建立CRF大鼠模型,连续造模21 d;模型复制成功后开始灌胃给药,每日1次,持续干预28 d。检测大鼠血清肌酐(SCr)、尿素氮(BUN)及24 h尿蛋白(24 h U-pro)水平;采用HE染色法及MASSON染色法检测大鼠肾脏组织病理变化,计算胶原容积分数(CVF);免疫组化法检测肾脏组织中PTEN、PI3K、p-AKT蛋白表达。结果 (1)网络药理学预测得到肾舒复健汤治疗CRF的活性成分140个,关键靶点(共同靶点)199个,核心活性化合物包括金合欢素、木犀草素、槲皮素等;核心靶点包括蛋白激酶B(AKT1)、肿瘤蛋白53(TP53)、信号转导及转录激活因子3(STAT3)等;GO功能主要与缺氧反应、血管生成正向调节、蛋白质磷酸化正调节等生物过程相关;关键通路主要涉及癌症途径、PI3K/AKT信号通路、Ras信号通路等;调控PTEN/PI3K/AKT信号通路可能是肾舒复健汤治疗CRF的重要作用机制之一。(2)体内试验显示,与正常组比较,模型组大鼠的血清SCr、BUN及24 h U-pro水平显著升高(P<0.01);肾脏组织病理改变明显,可见大量蓝色胶原纤维沉积,CFV明�Objective To investigate the mechanism of action of Shenshu Fujian Decoction(Radix Astragali,Rhizoma Atractylodis Macrocephalae and Radix Rehmanniae Recens)in the treatment of chronic renal failure(CRF)based on network pharmacology combined with in vivo experiments in animals. Methods(1)The active ingredients of Shenshu Fujian Decoction were searched and screened by TCMSP database, and the potential targets were screened by Swiss database;CRF disease-related targets were retrieved by DisGeNET and GeneCards databases;The intersection of potential targets of active ingredients and disease-related targets of CRF was obtained by Veen platform,and the key target(common target)of Shenshu Fujian Decoction for the treatment of CRF was obtained.Cytoscape 3.7.2 software was used to construct the "active ingredients-key target" network of Shenshu Fujian Decoction for the treatment of CRF,and the core active ingredients were analyzed;common targets were imported into STRING database, protein-protein interaction(PPI) network was constructed to screen core targets;the common targets were imported into DAVID database for GO function and KEGG pathway enrichment analysis.(2)Rats were randomly divided into normal group, model group, Niaoduqing Granules group(5 g · kg), and Shenshu Fujian Decoction group(11 g·kg^(-1)),with 8 rats in each group;CRF model was established by feeding0.5% adenine for consecutive 21 days. After successful replication of the model,gavage administration was started once daily for consecutive 28 days. Serum creatinine(SCr), blood urea nitrogen(BUN) and 24-hour urine protein(24 h U-pro) were detected;HE staining and MASSON staining were used to detect the pathological changes of kidney tissues and calculate the collagen volume fraction(CVF). The protein expressions of PTEN, PI3K and P-Akt in renal tissues were detected by immunohistochemistry. Result There were 140 key active ingredients and199 key targets(common targets)predicted for the treatment of CRF with Shenshu Fujian Decoction. The core active comp

关 键 词：肾舒复健汤 慢性肾衰竭 肾纤维化 网络药理学 PTEN/PI3K/AKT信号通路 大鼠 

分 类 号：R285.5[医药卫生—中药学] R857.3[医药卫生—中医学]