血清饥饿可诱导体外培养的血管平滑肌细胞(vascular smooth muscle cells,VSMC)由合成型转变为收缩型,微丝重塑是该过程的一个重要事件。平滑肌22α(smooth muscle 22 alpha,SM22α)是VSMC的标志蛋白,为了证实SM22α是否参与调节VSMC的微丝重塑过程,采用反义技术,封闭SM22α表达,利用间接免疫荧光染色、透射电镜观察SM22α表达对VSMC微丝重塑的影响,利用细胞平面迁移实验观察SM22α表达对VSMC运动功能的影响。实验结果显示,在血清饥饿培养的VSMC中,伴随着SM22α和SMα肌动蛋白表达上调,微丝数量明显增多,呈极性束状分布。用反义SM22α抑制SM22α表达后,血清饥饿诱导的VSMC微丝重塑受阻,微丝纤细,排列紊乱,且细胞迁移活性下降。结果提示,在VSMC微丝组装过程中,SM22α可能起一种捆绑蛋白作用。
Smooth muscle 22 alpha (SM22α) is a marker of vascular smooth muscle cells (VSMC). In order to investigate the effect of SM22α on microfilament remodeling, antisense SM22α was used in present study. The change of microfilaments in VSMC was observed by immunofluorescence and transmission electron microscopy (TEM), and the ability of migration of VSMC was measured by using a monolayer wounding model. The results demonstrated that the expression of SM22α and SM α-actin was induced quickly after serum starvation and the microfilaments were remodulated from discrete network to well arranged and dense bundles. However, after VSMC was transfected by antisense SM22α recombinant plasmid, reorganization of cytoskeleton by serum starvation was blocked, and the ability of migration of VSMC was declined. These data suggested that SM22α plays a key role during microfilament remodeling.