miR-381-3p靶向调控Sox9对高糖诱导的肾小管上皮细胞凋亡的影响  

Effect of miR-381-3p on high glucose-induced renal tubular epithelial cell apoptosis by targeting and regulating Sox9

作　　者：蒋双 李利娟[1] 乔芳 王麒智 JIANG Shuang;LI Lijuan;QIAO Fang;WANG Qizhi(Department of General Medicine,2.Department of General Surgery,Guang'an People's Hospital,Guang'an Sichuan 638000,China)

机构地区：[1]四川省广安市人民医院全科医学科,638000 [2]四川省广安市人民医院普外一科,638000

出　　处：《蚌埠医科大学学报》2025年第2期152-158,共7页Journal of Bengbu Medical University

摘　　要：目的:探讨微小RNA-381-3p(miR-381-3p)靶向性别决定区Y框蛋白9(Sox9)对高糖(HG)诱导肾小管上皮细胞(HK-2)凋亡的影响。方法:体外培养肾小管上皮细胞系HK-2细胞,将HK-2细胞分为对照组、HG组、HG+miR-NC组(转染miR-NC)、HG+miR-381-3p mimics组(转染miR-381-3p mimics)、HG+si-NC组(转染si-NC)、HG+si-Sox9组(转染si-Sox9)、HG+miR-381-3p mimics+pcDNA组(miR-381-3p mimics与pcDNA共转染)和HG+miR-381-3p mimics+pcDNA-Sox9组(miR-381-3p mimics与pcDNA-Sox9共转染),除对照组外,其余组均用30 mmol/L葡萄糖处理。实时荧光定量PCR(RT-qPCR)法检测HK-2细胞miR-381-3p及Sox9 mRNA表达;流式细胞仪检测细胞凋亡情况;免疫印迹法检测各组HK-2细胞Sox9、B淋巴细胞瘤-2相关蛋白(Bax)、B淋巴细胞瘤-2(Bcl-2)蛋白表达;双荧光素酶报告基因实验验证miR-381-3p与Sox9的靶向关系。结果:与对照组比较,HG组细胞凋亡率、Bax mRNA及蛋白表达水平升高,miR-381-3p、Bcl-2 mRNA及蛋白表达水平降低,差异均有统计学意义(P<0.05)。经targetscan数据库预测显示,miR-381-3p与Sox9 mRNA 3′UTR区有结合位点。过表达miR-381-3p或抑制Sox9表达,细胞凋亡率、Bax、Sox9 mRNA及蛋白表达水平降低,Bcl-2 mRNA及蛋白表达水平升高,差异均有统计学意义(P<0.05);过表达Sox9可逆转过表达miR-381-3p对HG诱导的HK-2细胞凋亡的抑制作用(P<0.05)。结论:miR-381-3p在HG诱导的肾小管上皮细胞中低表达,过表达miR-381-3p可通过靶向抑制Sox9表达,从而减少了HG诱导的肾小管上皮细胞凋亡。Objective:To investigate the effect of microRNA-381-3p(miR-381-3p)targeting SRY related HMG box-9(Sox9)on high glucose(HG)-induced renal tubular epithelial HK-2 cells apoptosis.Methods:The renal tubular epithelial HK-2 cells were cultured in vitro,and divided into control group,HG group,HG+miR-NC group(transfected with miR-NC),HG+miR-381-3p mimics group(transfected with miR-381-3p mimics),HG+si-NC group(transfected with si-NC),HG+si-Sox9 group(transfected with si-Sox9),HG+miR-381-3p mimics+pcDNA group(co-transfected with miR-381-3p mimics and pcDNA)and HG+miR-381-3p mimics+pcDNA-Sox9 group(co-transfected with miR-381-3p mimics and pcDNA-Sox9).Except the control group,the cells in other groups were treated with 30 mmol/L glucose.The expression of miR-381-3p and Sox9 at mRNA level in HK-2 cells was detected by real-time fluorescent quantitative PCR,the apoptosis was detected by flow cytometry,the protein expressions of Sox9,B lymphoma-2 related protein(Bax)and B lymphoma-2(Bcl-2)in HK-2 cells were detected by Western blotting,and the targeting relationship between miR-381-3p and Sox9 was verified by the dual luciferase reporter gene experiment.Results:Compared with the control group,the apoptosis rate,the mRNA and protein expression levels of Bax in HG group were increased,and the expression of miR-381-3p and Bcl-2 at mRNA and protein levels were decreased,which had significant statistical difference(P<0.05).The targetscan database predicted that miR-381-3p had a binding site with the 3'UTR region of Sox9 mRNA.When overexpression of miR-381-3p or inhibition of Sox9 expression,the apoptosis rate,the mRNA and protein expression levels of Bax and Sox9 were significantly reduced(P<0.05),and the mRNA and protein expression level of Bcl-2 was significantly increased(P<0.05).Overexpression of miR-381-3p or inhibition of Sox9 expression decreased the apoptosis rate,mRNA and protein expression levels of Bax and Sox9,and increased the mRNA and protein expression levels of Bcl-2,the difference was statistically significant(P<

关 键 词：微小RNA-381-3p 性别决定区Y框蛋白9 肾小管上皮细胞 凋亡 

分 类 号：R587.1[医药卫生—内分泌]