微RNA-181a-5p介导Sirt1/PPARγ信号通路促进肾病综合征大鼠细胞凋亡的机制研究  

Mechanism of apoptosis in rats with nephrotic syndrome by miR-181a-5p through Sirt1/PPARγsignaling pathway

作　　者：崔凯[1] 张睿[1] 栾仲秋[2] CUI Kai;ZHANG Rui;LUAN Zhongqiu(The First Clinical Medical College of Heilongjiang University of Chinese Medicine,Harbin,Heilongjiang 150040,China;Department of Nephropathy,The First Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin,Heilongjiang 150040,China)

机构地区：[1]黑龙江中医药大学第一临床医学院,黑龙江哈尔滨150040 [2]黑龙江中医药大学附属第一医院肾病一科,黑龙江哈尔滨150040

出　　处：《安徽医药》2024年第9期1756-1761,I0002,共7页Anhui Medical and Pharmaceutical Journal

基　　金：黑龙江省自然科学基金联合引导项目(LH2022H073)。

摘　　要：目的旨在探究微RNA(miR)-181a-5p对肾病综合征大鼠细胞凋亡的调控机制。方法该研究起止年限为2021年1月至2022年12月。使用阿霉素(ADR)构建肾病综合征大鼠模型及体外肾病综合征损伤模型,转染miR-181a-5p inhibitor/NC至体外细胞模型。使用生化分析仪分析血清肌酐、血尿素氮、血清白蛋白和尿蛋白,实时荧光定量逆转录聚合酶链反应(qRTPCR)检测肾组织及大鼠肾足细胞miR-181a-5p表达水平,通过TargetScan(http://www.targetscan.org/vert_72/)预测miR-181a-5p的下游靶基因,双萤光素酶报告实验来验证miR-181a-5p与沉默信息调节因子1(Sirt1)靶向结合关系。使用CCK-8试剂盒检测细胞增殖,流式细胞术检测细胞凋亡,通过蛋白质印迹法检测Sirt1及过氧化物酶体增殖物激活受体γ(PPARγ)蛋白水平。结果肾病综合征大鼠miR-181a-5p相对表达水平为3.50±0.30,敲低miR-181a-5p,使得Sirt1蛋白表达水平从0.36±0.02上调至0.87±0.06,PPARγ蛋白表达水平从0.26±0.02上调至0.78±0.05。结论miR-181a-5p通过抑制Sirt1/PPARγ信号通路活性促进肾足细胞凋亡。Objective To explore the regulatory mechanism of microRNA(miR)-181a-5p on apoptosis in rats with nephrotic syndrome.Methods The starting and ending years of the study were from January 2021 to December 2022.A rat model of nephrotic syndrome and an in vitro nephrotic syndrome injury model were constructed using adriamycin(ADR),and miR-181a-5p inhibitor/NC was transfected into the in vitro cell model.Serum creatinine,blood urea nitrogen,serum albumin and urinary protein were analyzed using a biochemical analyzer,and miR-181a-5p expression levels were detected in renal tissues and glomerular podocytes of rats by real-time fluorescence quantitative reverse transcription polymerase chain reaction(qRT-PCR).TargetScan(http://www.targetscan.org/vert_72/)was adopted to predict the downstream target genes of miR-181a-5p,and dual luciferase reporter assay to verify the target binding relationship between miR-181a-5p and silent information regulator 1(Sirt1).Cell proliferation was detected using CCK-8 kit,apoptosis by flow cytometry,and Sirt1 and peroxisome proliferator activated receptorγ(PPARγ)protein levels by protein blotting.Results The relative expression level of miR-181a-5p in nephrotic syndrome rats was 3.50±0.30.Knockdown of miR-181a-5p resulted in the up-regulation of Sirt1 protein expression level from 0.36±0.02 to 0.87±0.06,and the up-regulation of PPARγprotein expression level from 0.26±0.02 to 0.78±0.05.Conclusion miR-181a-5p promotes glomerular podocyte apoptosis by inhibiting Sirt1/PPARγsignaling pathway activity.

关 键 词：肾病综合征 肾足细胞 微RNA-181a-5p 沉寂信息调节因子 过氧化物酶体增殖物激活受体Γ 凋亡 阿霉素 大鼠 Sprague-Dawley 

分 类 号：R692[医药卫生—泌尿科学]