糖基化终产物通过微小RNA-30a调控自噬对血管内皮细胞功能的影响

Advanced glycation endproducts induce vascular endothelial dysfunction by regulating autophagy through miR-30a

目的探讨微小RNA(miR)-30a在糖基化终产物(AGE)调控自噬中的作用及其对血管内皮细胞功能的影响。方法将人主动脉内皮细胞(HAEC)随机分为对照组、AGE组、AGE+miR-30a对照组、AGE+miR-30a模拟组和AGE+miR-30a抑制剂组,检测内皮型一氧化氮合酶(eNOS)、微管相关蛋白轻链3(LC3)和Beclin-1 mRNA和蛋白表达;测定上清中血浆内皮素1及细胞间黏附分子1(ICAM-1)水平。结果与对照组比较,AGE组eNOS、LC3、Beclin-1 mRNA和蛋白表达及LC3斑点累积吸光度更低(P=0.000)。AGE组上清液NO水平明显低于对照组[(0.78±0.14)μg/L vs(4.82±0.65)μg/L,P=0.000],内皮素1、ICAM-1水平明显高于对照组[(451.96±86.39)pg/ml vs(69.13±11.67)pg/ml,P=0.000;(63.51±10.68)μg/ml vs(18.43±2.32)μg/ml,P=0.000]。与AGE+miR-30a对照组和AGE+miR-30a模拟组比较,AGE+miR-30a抑制剂组LC3、Beclin-1、eNOS mRNA和蛋白表达以及LC3斑点累积吸光度明显增加(P=0.000),NO水平明显升高,内皮素1和ICAM-1水平明显降低(P=0.000)。结论AGE通过上调miR-30a表达抑制HAEC自噬反应,诱导HAEC功能发生紊乱。

Objective To investigate the role of miR-30 a in autophagy regulated by AGE and its effect on the function of vascular endothelial cells.Methods HAEC were randomly divided into control group,AGE group,AGE+miR-30 a control group,AGE+miR-30 a mimics group,and AGE+miR-30 a inhibitor group.The mRNA expression was detected and the protein levels of eNOS,LC3,and Beclin-1 were measured.The ET-1 and ICAM-1 were detected.Results Compared with the control group,the mRNA and protein levels of eNOS,LC3,and Beclin-1,the cumulative optical density of LC3 spots(P=0.000),and the NO content in supernatant(0.78±0.14μg/L vs 4.82±0.65μg/L,P=0.000)were significantly decreased,while the concentrations of ET-1 and ICAM-1(451.96±86.39 pg/ml vs 69.13±11.67 pg/ml,P=0.000;63.51±10.68μg/ml vs 18.43±2.32μg/ml,P=0.000)were obviously increased in AGE group.The AGE+miR-30 a inhibitor group had significantly increased mRNA and protein levels of eNOS,LC3,and Beclin-1,higher cumulative optical density of LC3 spots,elevated NO concentration,but decreased ET-1 and ICAM-1 concentrations in supernatant when compared with AGE+miR-30 a control group and AGE+miR-30 a mimics group(P=0.000).Conclusion AGE inhibits the autophagy of HAEC by up-regulating miR-30 a,and finally induces the dysfunction of HAEC.