层流剪切力作用下miR-21对内皮细胞Caveolin-1基因表达的影响  

Effect of miR-21 on expression of Caveolin-1 gene in endothelial cells under laminar shear stress

作　　者：郭大伟[1] 张晓帆[1] 刘振东[1] 

机构地区：[1]山东省医学科学院基础医学研究所,山东济南250062

出　　处：《生物医学工程与临床》2017年第4期421-424,共4页Biomedical Engineering and Clinical Medicine

基　　金：国家自然科学基金项目(81670432);山东省自然科学基金项目(ZR2014HM098)

摘　　要：目的探讨层流剪切力(SS)作用下miR-21对内皮细胞Caveolin-1基因表达的影响。方法采用Lipofectamine2000将miR-21 mimics和miR-21 inhibitor分别转染到人主动脉内皮细胞,将转染后的内皮细胞分别加载低[0.4 Pa(4 dyn/cm^2)]和高[1.5 Pa(15 dyn/cm^2)]SS,并作用24 h。采用逆转录聚合酶链反应(RT-PCR)及Western blot技术检测Caveolin-1 m RNA和Caveolin-1蛋白的表达。结果对照组、低SS+miR-21 mimics组、低SS+miR-21 inhibitor组、高SS+miR-21 mimics组及高SS+miR-21 inhibitor组Caveolin-1 mRNA/甘油醛-3-磷酸脱氢酶(GAPDH)表达分别为1.08±0.17、0.44±0.16、1.19±0.34、0.93±0.27及1.70±0.39,Caveolin-1/β-actin蛋白分别为1.02±0.24、0.44±0.24、0.98±0.34、1.01±0.35及1.51±0.48。低SS+miR-21 mimics组Caveolin-1蛋白表达显著低于对照组(P<0.05),高SS+miR-21 inhibitor组Caveolin-1蛋白表达显著高于其他4组(P<0.05),低SS+miR-21 inhibitor组及高SS+miR-21 mimics组Caveolin-1蛋白表达显著高于低SS+miR-21 mimics组(P>0.05)。结论 miR-21对层流SS作用下内皮细胞的Caveolin-1基因的表达可能具有重要的调控作用。Objective To investigate the effect of miR-21 on the expression of Caveolin-1 gene in endothelial cells under laminar shear stress（SS）. Methods The cultural of human aortic endothelial cells（HAEC） were transfected by miR-21 mimics and miR-21 inhibitor by Lipofectamine 2000. The low[0.4 Pa（4 dyn/cm^2）] and high[1.5 Pa（15 dyn/cm^2）] SS was exerted on transfected HAEC for 24-hour, respectively. The expressions of Caveolin-1 m RNA and Caveolin-1 protein were assessed by reverse transcription polymerase chain reaction（RT-PCR） and Western Blot, respectively. Results The Caveolin-1 m RNA/glyceraldehyde-3-phosphate dehydrogenase（GAPDH） expression in control group, low SS ＋ miR-21 mimics group, low SS ＋ miR-21 inhibitor group, high SS ＋ miR-21 mimics group and high SS ＋ miR-21 inhibitor group was 1.08 ± 0.17, 0.44 ± 0.16, 1.19 ± 0.34,0.93 ± 0.27, and 1.70 ± 0.39, respectively. The expressions of Caveolin-1/β-actin protein were 1.02 ± 0.24, 0.44 ± 0.24, 0.98 ±0.34, 1.01 ± 0.35 and 1.51 ± 0.48, respectively. The Caveolin-1 protein expression in low SS ＋ miR-21 mimics group was significantly lower than that of control group（P 〈 0.05）, Caveolin-1 protein expression in high SS ＋ miR-21 inhibitor group was significantly higher than that of other 4 groups（P 〈 0.05）, and Caveolin-1 protein expressions in low SS ＋ miR-21 inhibitor group and high SS ＋ miR-21 mimics group were significantly higher than that of low SS ＋ miR-21 mimics group（P 〈 0.05）. Conclusion It is demonstrated that miR-21 may have an important regulation effect on the expression of Caveolin-1 gene in endothelial cells under laminar SS.

关 键 词：microRNA CAVEOLIN-1基因 CAVEOLIN-1蛋白 剪切力 内皮细胞 小窝蛋白 质膜小窝 

分 类 号：R318.01[医药卫生—生物医学工程]