摘要
目的探讨微小RNA-27a(miR-27a)介导的雷帕霉素靶分子(mTOR)信号通路调控DKD大鼠足细胞自噬的作用。方法将30只SD大鼠随机分为正常对照组(NC,n=10)、糖尿病模型组(DKD,n=10)和雷帕霉素治疗组(Rap,n=10)。检测各组肾功能相关指标和细胞凋亡率,HE染色及免疫组化评估肾组织形态学变化。Western blot法检测各组肾组织中自噬相关指标Beclin1、微管相关蛋白1A/1B-轻链3(LC3)及mTOC蛋白表达,qRT-PCR检测足细胞特异性蛋白miR-27a、Podocalyxin、Nephrin、LC3-Ⅰ、LC3-Ⅱ、Bax、Beclinl mRNA表达量。结果与NC组比较,DKD组第6、12周FPG、24 hUAlb、BUN、血肌酐(Scr)、肾脏指数(KI)表达升高(P<0.05)。与NC组比较,DKD组细胞凋亡、mTOC蛋白、miR-27a、mTOR、Bax mRNA表达升高(P<0.05),Beclinl、LC3-Ⅰ、LC3-Ⅱ蛋白、LC3-Ⅰ、LC3-Ⅱ、Beclin1、Podocalyxin、Nephrin mRNA表达降低(P<0.05)。与DKD组比较,Rap组Beclinl、LC3-Ⅰ、LC3-Ⅱ蛋白、LC3-Ⅰ、LC3-Ⅱ、Beclinl、Podocalyxin、Nephrin mRNA表达升高(P<0.05),细胞凋亡、mTOC蛋白、miR-27a、mTOR、Bax mRNA表达降低(P<0.05)。结论DKD大鼠肾组织miR-27a过表达可能介导mTOR自噬信号通路损伤足细胞,雷帕霉素能下调miR-27a表达,抑制mTOR/Beclinl/LC3信号通路,保护肾小球足细胞。
Objective To investigate the role of microRNA27a(miR-27A)mediated mTOR signaling pathway in regulating podocyte autophagy in rapamycin diabetic nephropathy rats.Methods Thirty SD rats were randomly divided into normal control group(NC,n=10),model group(DKD,n=10)and rapamycin group(Rap,n=10).HE staining and immunohistochemistry were used to evaluate the changes of renal histomorphology.Cell apoptosis rate was detected.Beclinl,microtubule associated protein 1A/1b-light chain 3(LC3)and mTOC protein expressions in renal tissues of each group were detected by Western blot.mRNA expression levels of Podocalyxin,miR-27a,Nephrin,LC3-I,LC3-II,Bax and Beclinl were detected by qRT-PCR.Results Compared with NC group,FPG,24 hUAlb,BUN,serum creatinine(Scr)and renal index(KI)increased at the 6th and 12th week in DKD group(P<0.05).Compared with NC group,the apoptosis,the expression of mTOC protein,miR-27a,mTOR and Bax mRNA increased(P<0.05),while the expressions of Beclinl,LC3-I,LC3-II protein,LC3-I,LC3-II,Beclinl,Podocalyxin and Nephrin mRNA decreased in DKD group(P<0.05).Compared with DKD group,the mRNA expressions of Beclinl,LC3-I,LC3-II protein,LC3-II,LC-3II,Beclinl,Podocalyxin and Nephrin mRNA increased(P<0.05),while the mRNA expressions of apoptosis,mTOC protein,miR-27a,mTOR and Bax decreased in Rap group(P<0.05).CConclusion The overexpression of miR-27amay mediate the damage of mTOR autophagy signaling pathway to podocytes in renal tissue of DKD rats,and rapamycin can down-regulate the expression of miR-27a and inhibit mTOR/Beclinl/LC3 signaling pathway to protect glomerular podocytes.
作者
杜彦丽
袁捷
姜云生
王肃
DU Yanli;YUAN Jie;JIANG Yunsheng(Department of Endocrinology,Tianjin Fifth Central Hospital,Tianjin 300450,China)
出处
《中国糖尿病杂志》
CAS
CSCD
北大核心
2023年第8期609-614,共6页
Chinese Journal of Diabetes