摘要
目的研究miR-138-5p对高糖诱导的人肾小管上皮细胞间充质转化(EMT)的影响。方法将HK-2细胞分为正常组(NG,5mmol/L D-葡萄糖)、高糖组(HG,30mmol/L D-葡萄糖)、HG+miR-138-5p抑制剂转染组及HG+抑制剂对照转染组。RT-PCR检测高糖处理对miR-138-5p表达的影响;Western blot检测平滑肌肌动蛋白(α-SMA)、上皮钙黏素(E-cadherin)、同源盒蛋白A13(HOXA13)、骨形成蛋白7(BMP-7)、转化生长因子β1(TGF-β1)和Smad7等的表达;荧光素酶活性实验检测miR-138-5p与HOXA13的靶向关系。结果高糖刺激后,miR-138-5p表达升高约1.5倍(P<0.05),α-SMA表达升高1.8倍,而E-cadherin表达下降;高糖促进TGF-β1的表达,抑制HOXA13、BMP-7和Smad7的表达(P<0.05)。miR-138-5p抑制剂转染组α-SMA和E-cadherin的表达均接近NG组,且miR-138-5p抑制剂转染组TGF-β1表达下降约1.1倍,同时HOXA13、BMP-7和Smad7表达升高(P<0.05)。结论抑制MiR-138-5p表达可通过靶向调节HOXA13的表达,提高BMP-7和Smad7水平,抑制TGF-β1信号通路,进而抑制高糖诱导的HK-2细胞EMT的发生。
Objective To analyze the effect of miR-138-5 p on the high glucose induced epithelial to mesenchymal transition（EMT）in human renal proximal tubular epithelial cells.Methods The cultured HK-2 cells were divided into four groups：normal glucose group（5 mmol/L D-glucose,NG）,high glucose group（30 mmol/L D-glucose,HG）,HG＋miR-138-5p inhibitor transfection group and HG＋miR-inhibitor negative control group.The expression of miR-138-5 p was measured by Real-time PCR.The expression ofα-SMA,E-cadherin,HOXA13,BMP-7,TGF-β1,and Smad7 were measured by western blot.The target relationship between miR-138-5p and HOXA13 was measured by luciferase activity assay.Results After high glucoses timulation,the expression of miR-138-5p increased by approximately 1.5 folds andα-SMA increased by 1.8 folds,while the expression of E-cadherin reduced.High glucose stimulation also dramatically up-regulated the expression of TGF-β1,down-regulated the expression of HOXA13,BMP-7 and Smad7（P〈0.05）.However,after miR-138-5 p inhibitor transfection,bothα-SMA and E-cadherin returned to similarlevel as compared with NG group.Furthermore,miR-138-5 p inhibitor dramatically decreased the expression of TGF-β1 by 1.1 folds,and increasedthe expression of HOXA13,BMP-7 and Smad7（P〈0.05）.miR-138-5 p could target to HOXA13 to modulate its expression.ConclusionInhibition of miR-138-5p increased the expression of BMP-7 and Smad7,suppressed the TGF-β1 pathway by regulating the expression of HOXA13,and therefore inhibited high glucose induced EMT in HK-2 cells.
作者
王晓妮
朱兵
陈卓玥
张海雄
龙开平
WANG Xiaoni;ZHU Bing;CHEN Zhuoyue;et al(Medical School, Xijing College ,Xi＇ an 710123 ,China)
出处
《中国糖尿病杂志》
CAS
CSCD
北大核心
2018年第1期64-68,共5页
Chinese Journal of Diabetes
基金
陕西省教育厅专项科研项目(16JK2245)
西京学院2017年科研基金(XJ170107)
