摘要
目的探讨miR-92b-5p在高糖诱导的足细胞损伤或凋亡过程中可能发挥的作用,并揭示其对线粒体功能的作用机制。方法①体外培养足细胞,分为对照组(NG组)和高糖组(HG组),通过高通量测序技术筛选差异表达的miRNA。②为了观察miR-92b-5p对足细胞线粒体功能和足细胞凋亡的影响,将正常血糖条件下培养的足细胞分为NC mimics、miR-92b-5p mimics、NC inhibitor、miR-92b-5p inhibitor组。为了观察抑制miR-92b-5p表达能否通过改善线粒体功能减轻高糖诱导的足细胞凋亡,将足细胞分为NG组、HG组、HG+NC inhibitor组、HG+miR-92b-5p inhibitor组。采用实时荧光定量聚合酶链式反应(qRT-PCR)测定miR-92b-5p的表达,流式细胞术测定足细胞凋亡率,CCK-8法测定足细胞活力,Western blot测定凋亡相关蛋白和线粒体相关蛋白的表达。结果①通过高通量测序技术,筛选出16个差异表达的miRNA,其中,HG组miR-92b-5p的表达明显升高(P<0.05)。②与NC mimics组相比,miR-92b-5p mimics组miR-92b-5p表达升高(P<0.05),足细胞活力降低(P<0.05),足细胞凋亡率增加(P<0.05),Bcl-2、Mfn1表达降低(P<0.05),Bax、Fis1表达增加(P<0.05)。与NC inhibitor组相比,miR-92b-5p inhibitor组miR-92b-5p表达降低(P<0.05),足细胞活力升高(P<0.05),足细胞凋亡率降低(P<0.05),Bcl-2、Mfn1表达升高(P<0.05),Bax、Fis1表达降低(P<0.05)。③与NG组相比,HG组miR-92b-5p表达升高(P<0.05),足细胞活力降低(P<0.05),足细胞凋亡率增加(P<0.05),Bcl-2、Mfn1表达降低(P<0.05),Bax、Fis1表达增加(P<0.05)。与HG+NC inhibitor相比,HG+miR-92b-5p inhibitor组miR-92b-5p表达降低(P<0.05),足细胞活力升高(P<0.05),足细胞凋亡率减少(P<0.05),Bcl-2、Mfn1表达升高(P<0.05),Bax、Fis1表达降低(P<0.05)。结论在高糖条件下,miR-92b-5p的表达水平增加,而抑制miR-92b-5p的表达能够通过改善线粒体功能减轻高糖诱导的足细胞凋亡,从而延缓糖尿病肾病的进展。
Objective To investigate the possible role of miR-92b-5p in podocyte injury or apoptosis induced by hyperglycemia,and explore its effect on mitochondrial function and its molecular mechanism.Methods①The podocytes were cultured and divided into normal glucose(NG)group and high glucose(HG)group,and the differentially expressed miRNAs were screened using high-throughput RNA-sequencing(RNA-seq)technology.②In order to investigate the effect of miR-92b-5p on podocyte mitochondrial function and podocyte apoptosis,the podocytes under normal glucose condition were divided into NC mimics group,miR-92b-5p mimics group,NC inhibitor,and miR-92b-5p inhibitor group.In order to investigate the effect of miR-92b-5p on podocyte apoptosis and mitochondrial function,the podocytes were divided into NG group,HG group,HG+NC inhibitor group,and HG+miR-92b-5p inhibitor group.Real-time polymerase chain reaction(qRT-PCR)was used to determine the expression of miR-92b-5p,flow cytometry was used to measure the podocyte apoptosis rate,CCK-8 was used to measure the podocyte viability,and Western blot was used to measure the expressions of apoptosis-related proteins and mitochondrial-related proteins.Results①Through high-throughput sequencing technology,16 differentially expressed miRNAs were identified,among which the expression of miR-92b-5p in HG group was significantly increased(P<0.05).②Compared with NC mimics group,the expression of miR-92b-5p was increased in miR-92b-5p mimics group(P<0.05),the cell viability was decreased(P<0.05),the podocyte apoptosis rate was increased(P<0.05),the expressions of Bcl-2 and Mfn1 were decreased(P<0.05),and the expressions of Bax and Fis1 were increased(P<0.05).③Compared with NG group,the expression of miR-92b-5p was increased in HG group(P<0.05),the podocyte viability was decreased(P<0.05),the podocyte apoptosis rate was increased(P<0.05),the expressions of Bcl-2 and Mfn1 were decreased(P<0.05),and the expressions of Bax and Fis1 were increased(P<0.05).Compared with HG+NC inhibitor group,the ex
作者
闪玥
李文贤
陈彦霞
王婷
贺银习
SHAN Yue;LI Wenxian;CHEN Yanxia;WANG Ting;HE Yinxi(Department of Endocrinology,Second Hospital of Hebei Medical University,Shijiazhuang 050000,China;Department of Endocrinology,First Hospital of Zhangjiakou;Department of Orthopaedic Trauma,Third Hospital of Shijiazhuang)
出处
《山西医科大学学报》
CAS
2024年第8期985-993,共9页
Journal of Shanxi Medical University
基金
河北省自然科学基金青年科学基金项目(H2020206108)
河北省医学科学研究课题(20210151)。
