下调miR-150-5p调控PI3K/AKT信号通路抑制体外糖尿病肾病足细胞模型凋亡的机制研究  

Mechanism of Down-regulation of miR-150-5p Inhibit Apoptosis in Diabetic Nephropathy Podocyte Model in Vitro Through PI3K/AKT Signaling Pathway

作　　者：江帆[1] 李伟[1] 陈娟[1] 冯光球[1] 管频[1] 曾敏[1] JIANG Fan;LI Wei;CHEN Juan(Geriatric Center of Hainan Provincial People’s Hospital,Haikou,570311)

机构地区：[1]海南省人民医院(海南医学院附属海南医院)老年医学中心,海口570311

出　　处：《中国中西医结合肾病杂志》2021年第3期205-209,共5页Chinese Journal of Integrated Traditional and Western Nephrology

基　　金：国家自然科学基金资助项目(No.81760054)。

摘　　要：目的:探讨下调miR-150-5p调控PI3K/AKT信号通路对体外糖尿病肾病足细胞模型凋亡的影响和机制。方法:以小鼠足细胞MPC5作为研究对象,分成Control(空白对照)、Model(高糖处理)、Anti-NC(转染inhibitor control后高糖处理)、Anti-miR-150-5p组(转染miR-150-5p inhibitor后高糖处理),Realtime PCR方法测定细胞中miR-150-5p表达,MTT方法测定细胞增殖变化,PI单染法检测细胞周期变化,Annexin V-FITC/PI双染法检测细胞凋亡变化,Western blot法检测细胞中C-Caspase-3、cyclinD1、p-AKT、p-PI3K蛋白表达。PI3K/AKT抑制剂LY294002处理转染miR-150-5p inhibitor后的足细胞,给予高糖处理,检测细胞增殖、周期和凋亡变化。结果:与Control组比较,Model组细胞中miR-150-5p表达水平升高,细胞增殖能力下降,细胞凋亡率升高,细胞G1期比例升高,细胞中C-Caspase-3蛋白表达水平升高,cyclinD1蛋白表达水平下降,p-AKT/AKT、p-PI3K/PI3K表达水平下降。与Anti-NC组比较,Anti-miR-150-5p组细胞中miR-150-5p表达水平降低,细胞增殖能力升高,细胞凋亡率降低,细胞G1期比例降低,细胞中C-Caspase-3蛋白表达水平降低,cyclinD1蛋白表达水平升高,p-AKT/AKT、p-PI3K/PI3K表达水平升高。PI3K/AKT抑制剂LY294002逆转miR-150-5p inhibitor对高糖条件下足细胞增殖、周期和凋亡影响。结论:下调miR-150-5p通过促进PI3K/AKT信号通路激活减少体外糖尿病肾病足细胞模型细胞凋亡。Objective:To investigate the effect and mechanism of down-regulated miR-150-5p expression on podocyte apoptosis under the high-glucose condition in vitro through PI3K/AKT signaling pathway.Methods:Mouse podocyte MPC5 was used as the research object and divided into Control,Model(high glucose treatment),Anti-NC(high glucose treatment with inhibitor control),Anti-miR-150-5p group(High glucose treatment with miR-150-5p inhibitor).The real-time PCR method was used to measure miR-150-5p expression in cells,and the MTT method was used to measure cell proliferation changes.The cell cycle changes were determined using PI single staining,while the changes of apoptosis were detected by Annexin V-FITC/PI double staining.The protein level of C-Caspase-3,cyclinD1,p-AKT,and p-PI3K protein in the cell was measured using Western blot.Results:Compared to the Control group,the expression level of miR-150-5p in the Model group cells was increased,and the cell proliferation capacity was decreased,with increased apoptosis rate and the G1 phase ratio.Western blot showed that the C-Caspase-3 protein expression level was enhanced,while the cyclinD1,p-AKT/AKT,and p-PI3K/PI3K protein expression level was down-regulated.Compared with the Anti-NC group,the expression level of miR-150-5p in the cells of the Anti-miR-150-5p group was reduced,the cell proliferation capacity was increased.Meanwhile,both the apoptosis and the G1 phase ratio of the cells was reduced.C-Caspase-3 The protein expression level was decreased,with increased protein levels of the cyclinD1,p-AKT/AKT,and p-PI3K/PI3K protein expression levels.Importantly,PI3K/AKT inhibitor LY294002 reversed the effects of miR-150-5p inhibitor on podocyte proliferation,cycle,and apoptosis under high glucose conditions.Conclusion:Down-regulation of miR-150-5p could reduce the apoptosis of podocytes under the high-glucose condition through the activation of PI3K/AKT signaling pathway.

关 键 词：糖尿病肾病 PI3K/AKT信号通路 miR-150-5p 凋亡 足细胞 

分 类 号：R587.2[医药卫生—内分泌] R692.9[医药卫生—内科学]