miR-206对高磷诱导大鼠血管平滑肌细胞钙化的影响及机制  

Effect of miR-206 on high phosphorus-induced calcification in rat vascular smooth muscle cells

作　　者：邵娟[1] 李敏才[1] 吴基良[1] 

机构地区：[1]湖北科技学院,湖北咸宁437100

出　　处：《山东医药》2016年第21期10-12,共3页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81270355);湖北省自然科学基金资助项目(2014CFB211)

摘　　要：目的观察微小RNA-206(miR-206)对β-甘油磷酸钠(β-GP)诱导的大鼠血管平滑肌细胞(VSMCs)钙化的影响,并探讨其机制。方法取原代培养大鼠主动脉VSMCs并进行细胞鉴定。将VSMCs随机分为5组:空白对照组、模拟物阴性对照组、miR-206模拟物组、抑制物阴性对照组、miR-206抑制物组,用脂质体Lipo2000进行转染。转染后48 h,各组给予β-GP 10 mmol/L诱导VSMCs钙化。诱导4 d,茜素红染色镜下观察各组钙化情况,采用微量酶标法检测细胞碱性磷酸酶(ALP)活性,采用免疫荧光法检测细胞缝隙连接蛋白43(Cx43)表达。结果VSMCs转染后48 h,经β-GP诱导4 d,与阴性对照组比较,miR-206模拟物组钙化结节减少,miR-206抑制物组钙化结节增多;与阴性对照组比较,miR-206模拟物组ALP活性及Cx43表达降低(P均<0.05),miR-206抑制物组ALP活性及Cx43表达升高(P均<0.05)。结论 miR-206可能通过调控Cx43表达抑制β-GP诱导的大鼠VSMCs钙化。Objective To observe the effect of microRNA-206（ miR-206） on β-glycerophosphate（ β-GP）-induced calcification in rat vascular smooth muscle cells（ VSMCs）,and to investigate its mechanism. Methods Aortal VSMCs were primarily cultured and identified in vitro. VSMCs were divided into five groups： control group,mimics negative control group,miR-206 mimics group,inhibitor negative control group and miR-206 inhibitor group. Each group was transfected by lipofectamine2000. After 48-hour transfection,each group was stimulated with 10 mmol / Lβ-GP to induce calcification of VSMCs. Observing calcium deposition with Alizarin red staining while detecting the alkaline phosphate（ ALP） activity and the expression of connexin 43（ Cx43） respectively by Trace enzyme standard method and immunofluorescence method after induction for 4 days. Results After transfection 48 h,VSMCs were treated with β-GP for 4 days. Compared with the negative control group,the calcium deposition decreased in the miR-206 mimics group and increased in the miR-206 inhibitor group; the ALP activity and the expression of Cx43 decreased in the miR-206 mimics group and increased in the miR-206 inhibitor group（ all P〈0. 05）. Conclusions miR-206 may inhibit β-GP-induced calcification of VSMCs by regulating Cx43.

关 键 词：微小RNA-206 β-甘油磷酸钠 血管平滑肌细胞 钙化 大鼠 

分 类 号：R543[医药卫生—心血管疾病]