RhoA蛋白参与调节白介素8诱导的血管内皮细胞迁移

Role of RhoA Protein on Regulating the Migration of Vascular Endothelial Cells Induced by IL-8

目的研究RhoA蛋白在内皮细胞迁移过程中的作用。方法采用脂质体包绕法将RhoA野生型(RhoAwt)、稳定表达持续活化型(RhoA63L)、主导抑制型(RhoA188A)突变质粒转染进入血管内皮细胞,获得稳定表达的EA.hy926-RhoAwt、EA.hy926-RhoA63L和EA.hy926-RhoA188A细胞。然后,利用Westernblot检测细胞中活性RhoA蛋白含量。最后,利用化学趋化因子IL-8(100ng/ml)分别刺激这3组细胞和对照组细胞,观察细胞迁移情况,并分析细胞迁移与活性RhoA之间的关系。结果1)各组细胞活性RhoA含量较对照组均有变化,EA.hy926-RhoAwt组稍稍升高,EA.hy926-RhoA63L组明显升高,而EA.hy926-RhoA188A组明显降低;2)在IL-8刺激下,EA.hy926-RhoAwt组细胞迁移能力稍高于对照组,EA.hy926-RhoA63L组细胞迁移能力远高于对照组,而EA.hy926-RhoA188A组细胞迁移能力则被明显抑制。因此,在IL-8刺激下,各组细胞迁移能力与细胞活性RhoA蛋白含量正相关。结论RhoA蛋白是IL-8诱导内皮细胞迁移过程中的关键分子,这有助于我们进一步探讨IL-8诱导内皮细胞迁移的理论机制和临床应用。

Objective To investigate the role of RhoA Protein in the migration of endothelial cell. Methods Firstly, endothelial cells were transfected to obtain the wild type RhoA（ RhoAwt）, the constitutively active forms of RhoA（ RhoA63L）, and the dominant negative forms of RhoA （ RhoA188A）, respectively, with lipofectamine 2000 reagent. Secondly, with Western blot technology, the content of activated RhoA in each type of these cells and the control group cells was measured respectively. Finally, with stimulation of IL-8 on these cells, the ability of cell migration was detected, and the interaction between cell migration and activated RhoA was analyzed. Results 1） The content of activated RhoA in each infected group was different with that in the control group. Content of EA. hy926-RhoAwt group was little higher, while EA. hy926-RhoAd3L group was great higher, however, EA. hy926-RhoA188A group was lower. 2） The ability of cell migration of EA. hy926-RhoAwt group was little higher, EA. hy926-RhoAd3L was great higher, and EA. hy926-RhoAlggA was inhibited ob- viously. The relation between ability of cell migration and content of activated protein RhoA was positive. Conclusion It is suggested that RhoA is a key factor in cellular signal transduction process of IL-8 inducing endothelial cell migration. These results will be helpful for investigating the mechanism of IL-8 inducing endothelial cell migration, and also for clinical application.