摘要
目的 探讨纤粘连蛋白介导的血管平滑肌细胞粘附和迁移与粘着斑激酶 (focaladhesionkinase ,FAK)的磷酸化的关系。方法 不同浓度的纤粘连蛋白 (fibronectin ,FN)刺激培养的血管平滑肌细胞 (smoothmusclecells,SMCs) ,观察细胞粘附反应 ,统计铺展比率。免疫沉淀和Wsternblot分别检测FAK及FAK磷酸化的表达量。利用改良的BoydenChamber测SMCs迁移。结果 FN有效地促进了SMC粘附 ,其铺展比率、迁移细胞数均显著高于对照 (P <0 0 5 ) ,且随FN浓度递增而增加。其中 2 0、40、6 0 μg ml组分别为 (75 6± 6 5 ) %、(80 9± 5 4) %和 (82 4± 7 9) % ,无组间差异 ,但均高于 5 μg ml的 (2 0 8± 3 2 ) %和 10 μg ml组的 (32 8± 4 7) % ,各组迁移细胞数也从 16 8± 3 6 HFP 2 0 0×增加到48 9± 6 1 HFP 2 0 0×。不同浓度FN作用后均有FAK的表达 ,FN10 μg ml即可致FAK磷酸化。表明FN介导SMCs粘附和迁移时伴有显著的FAK活化。结论 FN诱导平滑肌细胞粘附和迁移可能是通过FAK介导的 ,对其活性进行调控将有助于抑制血管损伤后内膜平滑肌细胞的迁移。
To investigate the role of focal adhesion kinase (FAK) phosphorylation in smooth muscle cells (SMCs) adhesion and migration mediated by fibronectin (FN) Methods Cultured rat aortic SMCs were stimulated by FN Cell adhesion and spreading were enumerated by light microscope Content of FAK and tyrosine kinase were investigated by immunoprecipitation and immunoblot Cell counting for migration were enumerated in modified Boyden Chamber Results We observed SMCs adhesion and spreading stimulated by FN significantly than the control Cell migration were also enhanced significantly In the mean time, expression of FAK were detected and tyrosine phosphorylation of FAK stimulated by at least 10 μg/ml FN were also observed It showed that tyrosine kinase activity was associated with SMCs adhesion and migration mediated by FN Conclusion SMCs adhesion and migration induced by FN maybe mediated by FAK To modulate tyrosine kinase activity of FAK may contribute to inhibit SMCs migration in injured arteries
出处
《中国介入心脏病学杂志》
2000年第3期149-151,共3页
Chinese Journal of Interventional Cardiology
关键词
血管平滑肌细胞
细胞外基质
粘着斑激酶
Vascular smooth muscle cells
Extracellular matrix
Focal adhesion kinase
Adhesion
Migration
