论文部分内容阅读
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，用相应引物进行转染。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（Cx-43）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 investigate the effect of microRNA-206 (miR-206)on β-glycerophosphate (β-GP)-in-duced calcification in rat vascular smooth muscle cells (VSMCs).Methods Aortal VSMCs were primarily cultured and i-dentified in vitro.VSMCs were divided into five groups:control group,mimics negative control group,miR-206 mimic group,inhibitor negative control group and miR-206 inhibitor group.Each group was transfected by lipofectamine2000.Af-ter 48-hour transfection,each group was stimulated with 10 mmol/Lβ-GP to induce calcification of VSMCs.Observing cal-cium deposition with Alizarin red staining while detecting the alkaline phosphate (ALP)activity and the expression of con-nexin 43 (Cx-43)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 mimic group and increased in the miR-206 inhibitor group;the ALP activity and the expression of Cx43 decreased in the miR-206 mimic 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.