miR-145与MyoCD调控VSMCs脂质蓄积的研究

Research of myocardin combined with miR-145 in regulating lipid accumulation in VSMC

目的:研究miR-145与心肌素(MyoCD)对人主动脉血管平滑肌细胞(VSMCs)脂质蓄积的影响。方法:ox-LDL孵育细胞,油红O染色观察细胞成脂,ELISA检测细胞内胆固醇含量;miR-145 siRNA及MyoCD过表达观察miR-145和MyoCD对血管平滑肌细胞脂质蓄积的影响,荧光定量PCR检测三磷酸腺苷结合盒转运体(ABCA)1的表达。结果:与对照组相比,MyoCD过表达组MyoCD蛋白及mRNA表达量均明显升高(P<0.05),miR-145 inhibitor组miR-145表达量下调(P<0.05)。ELISA检测结果表明,与空载+miR-145 inhibitor组相比,miR-145通过MyoCD过表达降低胆固醇含量。油红O染色结果显示,与空载组相比,抑制miR-145以及MyoCD+miR-145 inhibitor组均可使细胞内脂滴数量较少。PCR结果显示,抑制miR-145对ABCA1水平无显著影响。结论:miR-145低表达和MyoCD过表达可抑制VSMC细胞脂质蓄积。

Objective To explore effect of Myocardin(MyoCD)and miR-145 on regulating lipid accumulation in human aortic vascular smooth muscle cells(VSMC).Method VSMC was incubated with ox-LDL.Adipogenesis was observed by oil red O staining.Cholesterol content was detected by ELISA.The effect of miR-145 and MyoCD on lipid accumulation was detected by miR-145 siRNA and MyoCD overexpression vector.The expression of adenosine triphosphate binding cassette transporter 1(ABCA1)was detected by fluorescence quantitative PCR.Results Compared with the blank control group,the expression of MyoCD protein and mRNA in MyoCD overexpression group was significantly increased(P<0.05),and the expression of miR-145 in miR-145 inhibitor group was decreased(P<0.05).Oil red O staining showed that compared with the control group,the number of lipid droplets in miR-145 inhibitor group and MyoCD+miR-145 inhibitor group was less.Compared with the control group,the content of cholesterol in MyoCD+miR-145 inhibitor group decreased most(P<0.05).Overexpression of MyoCD and miR-145 inhibitor had no effect on the expression of ABCA1.Conclusion MyoCD overexpression combined with miR-145 inhibitor could inhibit lipid accumulation in VSMC cells.