阿托伐他汀对肌细胞增强因子2A基因突变型血管平滑肌细胞的作用

Effect of atorvastatin on myocyte enhancer factor 2A gene mutational vascular smooth muscle cells

目的观察阿托伐他汀对肌细胞增强因子2A(MEF2A)基因突变型血管平滑肌细胞(VSMC)的作用。方法将人主动脉血管平滑肌细胞分为3组:①WT组,转染野生型(WT)MEF2A质粒;②△21组,转染21碱基缺失突变型(△21)MEF2A质粒;③阿托伐他汀组,转染MEF2A△21质粒,并在实验前向该组细胞培养基中加入阿托伐他汀溶液至终浓度100μmol.L-1。通过溴化噻唑基蓝四唑(MTT)法和Millicell小室观察VSMC的增殖和迁移变化,免疫印迹(Western blotting)检测VSMC内平滑肌α肌动蛋白、SM22α、骨桥蛋白、p38和ERK1/2丝裂素活化蛋白激酶(MAPK)信号通路表达差异。结果 MEF2A基因△21突变可促进VSMC增殖、迁移和表型转化。而阿托伐他汀可抑制这些变化,并且明显下调磷酸化p38和ERK1/2信号通路表达。结论阿托伐他汀通过作用于p38和ERK1/2MAPK信号通路,以抑制MEF2A基因突变诱导的VSMC增殖、迁移和表型转化。

Objective To observe the effect of atorvastatin on myocyte enhancer factor 2A（MEF2A） gene mutational vascular smooth muscle cells（VSMC）.Methods The human aortic VSMC was randomly divided into 3 groups transfected with different plasmids：a WT group with MEF2A wild-type（WT） plasmid,a △21 group with MEF2A 21-base pair deletion mutation（△21） plasmid,and an atorvastatin group with MEF2A △21 plasmid and atorvastatin solution（100 μmol·L-1） added into the medium before the experiment.VSMC proliferation was determined by methyl thiazolyl diphenyl tetrazolium bromide（MTT）,and migration was measured by Millicell chamber.The expression of α-SM-actin,SM22α,osteopontin,p38,and ERK1/2 mitogen-activated protein kinase（MAPK） signaling pathway were detected by Western blot.Results MEF2A △21 improved the proliferation,migration,and phenotypic modulation of VSMC,but atorvastatin inhibited these changes.The phosphorylated p38 and ERK1/2 MAPK signaling pathway expression was significantly reduced by atorvastatin.Conclusion Through antagonizing p38 and ERK1/2 MAPK signaling pathway,atorvastatin can inhibit the proliferation,migration,and phenotypic modulation of vascular smooth muscle cells induced by MEF2A gene mutation.