摘要
目的探讨Racl活化在血小板衍生生长因子（PDGF-BB）刺激引起的大鼠主动脉平滑肌细胞增殖与迁移中的作用。方法贴块法分离培养主动脉平滑肌细胞，CCK8法和Transwell小室检测Racl抑制剂NSC23766和RaclsiRNA对PDGF-BB诱导的平滑肌细胞增殖和迁移的影响。GST．pulldown法和免疫印迹（Western印迹）检测PDGF-BB对Racl活性和pi-JNK表达的时间特性以及NSC23766和RaclsiRNA对Racl活性和pi—JNK表达的影响。结果PDGF—BB（50肛g／L）显著促进了大鼠主动脉平滑肌细胞的增殖和迁移。在给予不同浓度NSC23766（25、50、100¨g／L）以及RaclsiRNA（50nmol／L）后，其增殖和迁移显著受到了抑制。PDGF—BB刺激后Racl活性和pi．JNK逐渐升高，并分别在5min和15min达到最高峰后下降。给予NSC23766和RaclsiRNA处理后Racl活性（5min）和pi—JNK（15min）表达均明显受到了抑制。结论Racl活性影响JNK磷酸化并在调节PDGF—BB诱导的主动脉平滑肌细胞增殖与迁移中起着重要的作用。
Objective To explore the impact of Racl activation on the proliferation and migration under the stimulation of PDGF-BB （ platelet derived growth factor-BB）. Methods The inhibitory effects of Racl inhibitor （NSC23766） and RaclsiRNA on the proliferation and migration of vascular smooth muscle cell under the stimulation of PDGF-BB were measured by CCK8 assay and Transwell chamber. The time characteristics of Racl activity and pi-JNK expression under the stimulation of PDGF-BB was detected by GST pulldown assay and Western blot. And the inhibitory effects of NSC23766 and RaclsiRNA on the Racl activation and pi-JNK expression were also measured. Results Migration and proliferation of vascular smooth muscle cell increased significantly after the stimulation of PDGF-BB （ 50 trg/L） . Migration and proliferation was inhibited significantly after a pretreatment of RaclsiRNA and various concentrations of NSC23766（25, 50, 100μg/L）. After the stimulation of PDGF-BB, the expression of pi-JNK and Racl activity increased over time. Racl-GTP peaked at 5 minutes and pi-JNK at 15 minute. The expressions of pi- JNK at 15 minutes and Racl-GTP at 5 minutes were inhibited significantly by RaclsiRNA and NSC23766 in a concentration-dependent manner. Conclusion JNK phosphorylation is controlled by Racl activation. And Racl activation play a pivotal role in the migration and proliferation of aortic smooth muscle cell under the stimulation of PDGF-BB.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2011年第39期2793-2796,共4页
National Medical Journal of China
关键词
平滑肌细胞
血小板源性生长因子-BB
Rac1活性
增殖
迁移
Smooth muscle cell
Platelet derived growth factor-BB（PDGF-BB）
Racl activity
Proliferation
Migration
