CircACTR2调节miR-101-3p/NFAT5轴对高糖诱导的肾小管上皮细胞增殖、凋亡和上皮间质转化的影响  

Effects of circACTR2 on high glucose-induced cell proliferation,apoptosis and epithelial-mesenchymal transition in renal tubular epithelial cells by regulating miR-101-3p/NFAT5 axis

作　　者：王晶[1] 黄斌芳 周光全[1] WANG Jing;HUANG Binfang;ZHOU Guangquan(Department of Endocrinology and Nephrology,Hanyang Hospital Affiliated to Wuhan University of Science and Technology,Wuhan Hubei 430000,China)

机构地区：[1]武汉科技大学附属汉阳医院内分泌肾病科,湖北武汉430000

出　　处：《蚌埠医科大学学报》2025年第3期281-287,共7页Journal of Bengbu Medical University

摘　　要：目的:探讨环状非编码RNA(circRNA)ACTR2调节微小RNA-101-3p(miR-101-3p)/活化T细胞核因子5(NFAT5)轴对高糖诱导的肾小管上皮细胞增殖、凋亡和上皮间质转化的影响。方法:体外培养人肾小管上皮细胞HK-2,分为对照组、高糖组(30 mmol/L葡萄糖),同时在高糖组的基础上对HK-2细胞进行转染,设置为小干扰RNA阴性对照(si NC)组、circACTR2特异性小干扰RNA(si circACTR2)组、si circACTR2+抑制物阴性对照(inhibitor NC)组、si circACTR2+miR-101-3p抑制物(miR-101-3p inhibitor)组。流式细胞术、CCK-8分别检测细胞凋亡和增殖;qRT-PCR检测细胞中circACTR2、miR-101-3p及NFAT5 mRNA表达;Western blotting检测各组细胞中增殖蛋白Ki-67、抗凋亡蛋白Bcl-2、E-钙黏附蛋白(E-cadherin)、α-平滑肌肌动蛋白(α-SMA)、NFAT5表达水平;ELISA检测细胞中白细胞介素6(IL-6)、肿瘤坏死因子-α(TNF-α)水平;双荧光素酶实验验证circACTR2、NFAT5分别与miR-101-3p的靶向关系。结果:miR-101-3p分别与circACTR2、NFAT5存在靶向关系。与对照组相比,高糖组细胞凋亡率、TNF-α、IL-6、circACTR2、NFAT5、α-SMA水平增加(P<0.05),细胞光密度(OD_(450))值(24、48 h)、miR-101-3p、Bcl-2、Ki-67、E-cadherin水平降低(P<0.05);与si NC组相比,si circACTR2组细胞凋亡率、TNF-α、IL-6、circACTR2、NFAT5、α-SMA水平下降(P<0.05),细胞OD_(450)值(24、48 h)、miR-101-3p、Bcl-2、Ki-67、E-cadherin水平增加(P<0.05);与si circACTR2+inhibitor NC组相比,si circACTR2+miR-101-3p inhibitor组细胞凋亡率、TNF-α、IL-6、circACTR2、NFAT5、α-SMA水平增加(P<0.05),细胞OD_(450)值(24、48 h)、miR-101-3p、Bcl-2、Ki-67、E-cadherin水平降低(P<0.05)。结论:干扰circACTR2表达可以促进高糖诱导HK-2细胞增殖、抑制其凋亡及上皮间质转化,可能与调控miR-101-3p/NFAT5轴有关。Objective:To investigate the effects of circular non-coding RNA(circRNA)ACTR2 on high glucose-induced cell proliferation,apoptosis and epithelial-mesenchymal transition in renal tubular epithelial cells by regulating microRNA-101-3p(miR-101-3p)/nuclear factor of activated T cells 5(NFAT5)axis.Methods:Human renal tubular epithelial cells HK-2 were cultured in vitro and randomly divided into control group and high glucose group(30 mmol/L glucose).The cells in the high glucose group were transfected and set as small interfering RNA negative control(si NC)group,circACTR2-specific small interfering RNA(si circACTR2)group,si circACTR2+inhibitor negative control(inhibitor NC)group,and si circACTR2+miR-101-3p inhibitor(miR-101-3p inhibitor)group.Flow cytometry and CCK-8 were used to detect apoptosis and proliferation,respectively;qRT-PCR was applied to determine the mRNA expression of circACTR2,miR-101-3p and NFAT5 in cells;Western blotting was employed to analyze the expression levels of proliferation protein Ki-67,anti-apoptotic protein Bcl-2,E-cadherin,α-smooth muscle actin(α-SMA)and NFAT5;ELISA was performed to detect the level of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in cells;the dual luciferase assay was used to verify the targeted relationships of circACTR2 and NFAT5 with miR-101-3p.Results:MiR-101-3p had targeted relationships with circACTR2 and NFAT5.Compared with the control group,the apoptosis rate,TNF-α,IL-6,circACTR2,NFAT5 andα-SMA levels in the high glucose group increased(P<0.05),while cell optical density(OD_(450))values(24,48 hours),miR-101-3p,Bcl-2,Ki-67,and E-cadherin levels decreased(P<0.05).Compared with the si NC group,the apoptosis rate,TNF-α,IL-6,circACTR2,NFAT5 andα-SMA levels in the si circACTR2 group decreased(P<0.05),while cell OD_(450) values(24,48 hours),miR-101-3p,Bcl-2,Ki-67 and E-cadherin levels increased(P<0.05).Compared with the si circACTR2+inhibitor NC group,the apoptosis rate,TNF-α,IL-6,circACTR2,NFAT5 andα-SMA levels in the si circACTR2+miR-101-3p inhibitor

关 键 词：糖尿病肾病 环状非编码RNA ACTR2 微小RNA-101-3p/活化T细胞核因子5轴 肾小管上皮细胞 

分 类 号：R587.2[医药卫生—内分泌]