miR-194-5p通过靶向TLR4/NF-κB通路调控免疫因子IL-6/10对肾病综合征大鼠细胞凋亡的影响  

The effect of miR-194-5p on nephrocyte apoptosis in rats with nephrotic syndrome by targeting TLR4/NF-κB pathway and regulating IL-6/10

作　　者：陈义忠 何毅辉[2] 陈琅[3] 缪长新 郑鑫[3] CHEN Yizhong;HE Yihui;CHEN Lang;MIAO Changxin;ZHENG Xin(Basic Medical College,Fujian Medical University,Fuzhou 350122,China;Department of Pathology,Fujian Provincial Hospital&Fujian Medical University Provincial Clinical College,Fuzhou 350001,China;Department of Pediatrics,Fujian Provincial Hospital&Fujian Medical University Provincial Clinical College,Fuzhou 350001,China;Department of Pediatrics,Shouning County Hospital,Ningde 355500,China)

机构地区：[1]福建医科大学基础医学院,福州350122 [2]福建医科大学省立临床学院福建省立医院病理科,福州350001 [3]福建医科大学省立临床学院福建省立医院儿科,福州350001 [4]寿宁县医院儿科,宁德355500

出　　处：《免疫学杂志》2022年第2期116-123,共8页Immunological Journal

基　　金：福建省卫生健康科技项目中青年骨干项目(2020GGB004)。

摘　　要：目的探究微小RNA(microRNA,miR)-194-5p通过靶向Toll样受体4(Toll-like receptor 4,TLR4)/核因子(nuclear factor,NF)-κB通路调控免疫因子(interleukin,IL)-6/10对肾病综合征(nephrotic syndrome,NS)大鼠肾细胞凋亡的影响。方法将通过嘌呤霉素氨基核苷构建的NS大鼠随机分为NS组、NS+miR-194-5p agomir组和NS+miR-194-5p antagomir组(n=15),通过腹腔内注射miR-194-5p agomir或antagomir(300μg)以提高或抑制miR-194-5p。检测各组大鼠肾功能指标、炎性因子、凋亡情况以及TLR4/NF-κB通路水平。通过双荧光素酶报告验证miR-194-5p和TLR4的靶向关系。将肾胚细胞293细胞分为miR-194-5p NC组和miR-194-5p mimic组,通过转染miR-194-5p mimic过表达miR-194-5p,通过RT-qPCR和Western blot检测各组TLR4 mRNA和蛋白水平。结果NS组的24 h尿蛋白、肌酐、凋亡指数、IL-6、IL-10、TLR4和NF-κB蛋白表达水平均显著高于健康组(P<0.05);与NS组比较,NS+miR-194-5p antagomir组大鼠的24 h尿蛋白、肌酐、凋亡指数、IL-6、TLR4和NF-κB蛋白表达水平均显著升高,IL-10水平显著降低(P<0.05);NS+miR-194-5p agomir组的24 h尿蛋白、肌酐、凋亡指数、IL-6、IL-10、TLR4和NF-κB蛋白表达水平均显著降低(P<0.05)。双荧光素酶报告结果表明miR-194-5p可以与TLR4靶向结合。miR-194-5p mimic组细胞中的TLR4 mRNA和蛋白表达水平显著低于miR-194-5p NC组(P<0.05)。结论miR-194-5p可通过靶向抑制TLR4/NF-κB通路减少免疫炎性因子的释放和抑制细胞凋亡,从而缓解肾组织损伤缓解NS。The aim of this study was to explore the effects of microRNA(miR)-194-5p on cell apoptosis in nephrotic syndrome(NS) rats by targeting Toll-like receptor 4(TLR4)/nuclear factor(NF)-κB pathway and regulating immune factors IL)-6/10.Firstly,NS rats constructed by puromycin aminonucleosides were randomly divided into NS group,NS+miR-194-5p agomir group and NS+miR-194-5p antagomir group(n=15).miR-194-5p agomir or antagomir(300 μg) was intraperitoneal injection to increase or inhibit miR-194-5p.Then the renal function indexes,inflammatory factors,apoptosis and TLR4/NF-κB pathway levels of rats in each group were tested;the targeting relationship between miR-194-5p and TLR4 was verified by dual luciferase report.Kidney germ cell 293 were recruited and divided into miR-194-5p NC group and miR-194-5p mimic group.The TLR4 mRNA and protein levels in each group were detected by RT-qPCR and Western blotting.Data showed that 24-h urine protein,creatinine,apoptosis index,IL-6,IL-10,TLR4 and NF-κB protein expression levels in the NS group were significantly higher than those in the healthy group(P <0.05).Compared with the NS group,the 24-h urine protein,creatinine,apoptosis index,IL-6,TLR4 and NF-κB protein expression levels of rats in the NS+miR-194-5p antagomir group were increased significantly,wihle IL-10 levels significantly decreased(P <0.05).The 24-h urine protein,creatinine,apoptosis index,IL-6,IL-10,TLR4 and NF-κB protein expression levels in the NS+miR-194-5p agomir group were significantly reduced(P <0.05).The dual luciferase report results indicated that miR-194-5p could target and bind with TLR4.Furthermore,the expression levels of TLR4 mRNA and protein in the miR-194-5p mimic group were significantly lower than those in the miR-194-5p NC group(P <0.05).In conclusion,miR-194-5p can reduce the release of immune inflammatory factors and inhibit cell apoptosis through targeting inhibition of TLR4/NF-κB pathway,thereby alleviating kidney tissue damage and alleviating NS.

关 键 词：肾病综合征 miR-194-5p TOLL样受体4 核因子-ΚB 凋亡 

分 类 号：R692.39[医药卫生—泌尿科学]