高糖通过miR-192调控CAV-1/EGF影响肾小球系膜细胞增殖、迁移及细胞外基质形成的机制研究  

Effects of high glucose on the proliferation and migration of mesangial cells and extracellular matrix formation by regulating the CAV-1/EGF through miR-192

作　　者：吴玮熙 刘帅辉 周赛君[1] 刘红岩 张睿 于珮[1] WU Weixi;LIU Shuaihui;ZHOU Saijun;LIU Hongyan;ZHANG Rui;YU Pei(NHC Key Laboratory of Hormones and Development,Tianjin Key Laboratory of Metabolic Diseases,Chu Hsien-I Memorial Hospital&Tianjin Institute of Endocrinology,Tianjin Medical University,Tianjin 300134,China)

机构地区：[1]天津医科大学朱宪彝纪念医院,天津市内分泌研究所,国家卫健委激素与发育重点实验室,天津市代谢性疾病重点实验室,300134

出　　处：《天津医药》2022年第10期1020-1025,共6页Tianjin Medical Journal

基　　金：天津市卫生健康委科技人才培育项目(RC20175);森美中华糖尿病科研基金(Z-2017-26-1902);天津市医学重点学科(专科)建设项目。

摘　　要：目的探讨高糖条件下miR-192对肾小球系膜细胞增殖、迁移及细胞外基质形成的影响及可能的作用机制。方法体外培养人肾小球系膜细胞并分为6组:正糖(NG,5.6 mmol/L葡萄糖)组、高糖(HG,25 mmol/L葡萄糖)组、正糖加miR-192模拟物(NG+mimics)组、高糖加miR-192模拟物(HG+mimics)组、正糖加miR-192抑制剂(NG+inhibitor)组、高糖加miR-192抑制剂(HG+inhibitor)组。干预24 h后,CCK-8法检测细胞增殖活性,划痕实验检测细胞迁移能力,实时荧光定量PCR(qPCR)技术检测miR-192、小窝蛋白1(CAV-1)、表皮生长因子(EGF)、1型胶原蛋白(COLⅠ)、纤连蛋白(FN)mRNA水平,Western blot检测相关蛋白表达。结果与NG组相比,HG组细胞增殖率和划痕愈合率增加,EGF、FN、COLⅠmRNA和蛋白表达升高,CAV-1 mRNA和蛋白表达降低(均P<0.05)。在NG组和HG组分别加入miR-192 inhibitor后,细胞增殖率和划痕愈合率减少,EGF、FN、COLⅠmRNA和蛋白表达降低,CAV-1 mRNA和蛋白表达升高(均P<0.05),HG组加入miR-192 inhibitor变化更加明显。在NG组和HG组分别加入miR-192 mimics后,细胞增殖率和划痕愈合率增加,EGF、FN、COLⅠmRNA和蛋白表达升高,CAV-1 mRNA和蛋白表达降低(均P<0.05)。结论高糖通过miR-192-CAV-1-EGF途径增加肾小球系膜细胞增殖和迁移,导致细胞外基质的形成。Objective To investigate the effect of miR-192 on proliferation,migration and extracellular matrix of mesangial cells under high glucose condition.Methods Human mesangial cells were cultured in vitro for 24 h and divided into six groups:the normal glucose concentration(NG,5.6 mmol/L glucose)group,the high glucose concentration(HG,25 mmol/L glucose)group,the normal glucose concentration+miR-192 mimics(NG+mimics)group,the high glucose concentration+miR-192 mimics(HG+mimics)group,the normal glucose concentration+miR-192 inhibitor(NG+inhibitor)group and the high glucose concentration+miR-192 inhibitor(HG+inhibitor)group.After 24 h of intervention,cell viability was detected by CCK-8 assay,the cell migration ability was detected by Scratch healing test,and expression levels of miR-192,caveolin 1(CAV-1),epidermal growth factor(EGF),collagen type 1(COLⅠ)and fibronectin(FN)mRNA and related proteins were detected by qPCR and Western blot assay.Results Compared with the NG group,the proliferation and migration rates were increased,the expression levels of EGF,FN and COLⅠmRNA and protein were increased,and the expression of CAV-1 mRNA and protein decreased in the HG group(all P<0.05).When miR-192 inhibitor was added to the NG group and the HG group,the cell proliferation and migration were decreased,the expression of EGF,FN and COLⅠmRNA and protein were decreased,while the expression levels of CAV-1 mRNA and protein were increased(all P<0.05).Changes were more significant in the HG group after the addition of miR-192 inhibitor.After adding miR-192 mimics to the NG group and the HG group,the cell proliferation and migration increased,expression levels of EGF,FN and COLⅠmRNA and protein increased,and the expression of CAV-1 decreased(all P<0.05).Conclusion High glucose increases the proliferation and migration of glomerular mesangial cells through miR-192-CAV-1-EGF pathway,leading to the formation of extracellular matrix and participating in the occurrence and development of diabetic nephropathy.

关 键 词：糖尿病肾病 肾小球系膜细胞 表皮生长因子 细胞外基质 细胞增殖 微小RNA-192 小窝蛋白1 

分 类 号：R737.9[医药卫生—肿瘤]