论文部分内容阅读
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。
Objective To investigate the relationship between fibronectin-mediated adhesion and migration of vascular smooth muscle cells and phosphorylation of focal adhesion kinase (FAK). Methods Cultured vascular smooth muscle cells (SMCs) were stimulated with different concentrations of fibronectin (FN), and the cell adhesion reaction was observed. The spreading ratio was calculated. Immunoprecipitation and Wsternblot were used to detect the phosphorylation of FAK and FAK respectively. Migration of SMCs measured using a modified BoydenChamber. Results FN effectively promoted SMC adhesion. The spreading ratio and the number of migrating cells were significantly higher than that of control (P <0.05), and increased with the increasing of FN concentration. (75 6 ± 6 5)%, (80 9 ± 5 4)% and (82 4 ± 7 9)% respectively in 20, 40 and 60 μg ml groups without any difference among the groups, all of which were higher than 5 The number of migrating cells in each group also increased from 16 8 ± 3 6 HFP 200 × to 48 9 ± 6 (2 0 8 ± 3 2)% μg / ml and (32 8 ± 4 7)% in 10 μg ml group 1 HFP 2 0 0 ×. After different concentrations of FN have FAK expression, FN10 μg ml can cause FAK phosphorylation. FN-mediated SMCs adhesion and migration associated with significant FAK activation. Conclusion FN-induced smooth muscle cell adhesion and migration may be mediated through FAK, and its regulation of activity will help to inhibit the migration of intimal smooth muscle cells after vascular injury.