血管损伤后酪氨酸激酶活性变化与血管紧张素Ⅱ介导的血管平滑肌细胞迁移  

Protein tyrosin kinases activation associated vascular injury and angiotensin Ⅱ-induced migration of smooth muscle cell

作　　者：景涛[1] 何国祥[1] 吴昊[1] 刘建平[1] 王耿[1] 冉擘力[1] 王海东[2] 

机构地区：[1]第三军医大学西南医院心内科,重庆400038 [2]第三军医大学西南医院胸心外科,重庆400038

出　　处：《重庆医学》2004年第1期21-23,共3页Chongqing medicine

摘　　要：目的　本研究旨在探讨蛋白酪氨酸激酶活性变化在血管紧张素Ⅱ (AngⅡ )介导的血管平滑肌细胞 (VSMC)迁移中的作用。方法　体外培养VSMC ,采用 [γ 3 2 P] ATP掺入法、改良Boyden′schamber法和细胞化学的方法 ,观察AngⅡ干预前后VSMC胞浆内PTK活性的变化 ,粘着斑、肌动蛋白纤维丝的动态组装变化以及迁移能力的变化 ,探讨AT1 R拮抗剂、AT2 R拮抗剂以及酪氨酸激酶抑制剂对上述观测指标的影响。结果　 (1 )AngⅡ可以剂量依赖性诱导VSMC内胞浆PTK激活 ;(2 )AngⅡ刺激后 ,VSMC粘着斑表达增强 ,肌动蛋白丝数量明显增加 ,纵向平行排列 ;(3)AngⅡ刺激后VSMC发生迁移 ;(4 )AT1 R拮抗剂CV 1 1 974可显著抑制上述生物学效应 ,AT2 R拮抗剂PD1 2 331 9对此无明显的作用 ;酪氨酸激酶抑制剂Genistein可显著但不能完全抑制上述生物学效应。结论　AngⅡ通过AT1 R介导调节VSMC内粘着斑和肌动蛋白丝动态组装 ,进而改变VSMC的迁移能力 ;酪氨酸激酶的激活参与AngⅡ介导的VSMC迁移的信号转导调控 ;由于完全抑制PTK活性尚不能完全抑制AngⅡ诱导的VSM Cs跨膜迁移 ,因此VSMCs迁移的调控可能还有其他的信号转导通路的参与。Objective To investgate the effect of protein tyrosine kinases(PTK) on AngⅡ induced migration of VSMCs,and to explore the mechanisms of neintimal formation associated vascular injruy.Methods VSMCs isolated from aortic media of Wistar rats and cultured by the modified explant method were adopted.In prersence and absence of AngⅡ,the cytoplasm PTK activity was analyzed by method of [γ 32 P] ATP incorporation and reorganization of focal adhesion and actin cytoskeleton of VSMCs were studied by immunocytochemistry technique,fluorocytochemistry technique.The migration assays were performed by a modified Boyden′s chamber.And the effects of AT 1R antagonist(CV 11974),AT 2R antagonist(PD123319),PTK inhibitor(Genistein)on aforementioned target were studied.Results (1)AngⅡ stimulated cytoplasm PTK activity of VSMCs in a concentration dependent manner.(2)Treatment with AngⅡ resulted in a substantial increase in the number of stress fibers and rearragngement of these structures into ordered parallel arrays.There was a corresponding increase in the number and size of vinculin positive FA associated with the ends of actin filaments.(3)VSMC migrate after stimulate by AngⅡ.(4)Pretreated with CV 11974 significantly inhibited above mentioned effect.PD123319 had no significant effect.Pretreated with Genistein partly reduced above mentioned effect.Conclusion The dynamic assemble and disassemble of FA and reorganization of actin cytoskeleton may be required for AngⅡ induced VSMC migration,and this effect is mediated by AT 1R.The activation of PTK involved in the signaling transduction of VSMCs migration.VSMCs migration induced by AngⅡ isn′t inhibited completely,while PTK activity is absolutely inhibited,it suggests there be other signaling transdution pathway existing in regulation VSMCs migration.

关 键 词：血管紧张素Ⅱ 血管平滑肌细胞 酪氨酸激酶 

分 类 号：R543[医药卫生—心血管疾病]