KCa3.1通道对增殖表型大鼠血管平滑肌细胞增殖和迁移的影响

目的探讨KCa3.1通道对增殖表型大鼠血管平滑肌细胞增殖和迁移的影响。方法组织贴壁法培养大鼠血管平滑肌细胞,采用光学显微镜、电镜和免疫细胞化学染色法观察初代和9代平滑肌细胞形态学特征。RT-PCR和免疫细胞化学染色法检测初代和9代平滑肌细胞KCa3.1通道mRNA和蛋白表达。MTT和Transwell法观察KCa3.1通道对平滑肌细胞增殖和迁移的影响。结果初代和9代平滑肌细胞分别表现出收缩表型和增殖表型特征,9代平滑肌细胞KCa3.1通道mRNA、蛋白表达水平、增殖和迁移能力显著高于初代细胞(P<0.01),KCa3.1通道阻断剂TRAM-34可显著抑制9代细胞的增殖和迁移(P<0.01),对初代细胞无明显影响。结论增殖表型血管平滑肌细胞KCa3.1通道表达增加可能促进了细胞的增殖和迁移。

Role of K(Ca)3.1 channel in proliferation and migration of rat vascular smooth muscle cells of the proliferative phenotype

Objective To investigate the role of KCa3.1 channel in the proliferation and migration of rat vascular smooth muscle cells of the proliferative phenotype.Methods Rat vascular smooth muscle cells(VSMCs) were cultured with tissue adhesion method.The morphological characteristics of the fist and ninth passages of VSMCs were observed with light and electron microscopy and immunocytochemistry.The expressions of KCa3.1 channel mRNA and protein in the cells were detected using RT-PCR and immunocytochemistry,respectively.MTT and transwell assay were employed to assess the effect of the KCa3.1 channel blocker TRAM-34 on the proliferation and migration of VSMCs.Results The first and ninth passages of VSMCs showed morphological characteristics of contractile and proliferative phenotypes,respectively.Compared with the first-passage cells,the ninth-passage VSMCs exhibited significantly increased KCa3.1 channel mRNA and protein expressions with enhanced cell proliferation and migration(P<0.01),which was inhibited by the application of TRAM-34(P<0.01).TRAM-34 produced no obvious effect on the first-passage VSMCs.Conclusion Upregulated expression of KCa3.1 channel can promote the proliferation and migration of rat VSMCs of the proliferative phenotype.