黏着斑激酶对血小板源性生长因子刺激的血管平滑肌细胞迁移和黏附的调控  

Focal adhesion kinase regulates migration and adhesion of vascular smooth muscle cells induced by platelet derived growth factor

作　　者：武衡[1] 钟杨[1] 汤显靖[1] 成志[1] 

机构地区：[1]南华大学附属第一医院神经内科,衡阳421001

出　　处：《中华老年心脑血管病杂志》2010年第7期648-651,共4页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

基　　金：湖南省卫生厅医药卫生科研课题(B2004-092)

摘　　要：目的探讨黏着斑激酶(FAK)信号分子在雷帕霉素抑制血小板源性生长因子(PDGF)诱导血管平滑肌细胞(VSMC)迁移、黏附中的调控作用。方法将培养的大鼠VSMC分为对照组、PDGF组、雷帕霉素+PDGF组(雷帕霉素组)和FAK反义寡核苷酸+PDGF组(FAK组)。用PDGF诱导VSMC的迁移和黏附,计数贴壁细胞;采用Boyden检测细胞迁移;采用RT-PCR、Western blot、免疫沉淀方法分别检测FAK基因、蛋白及蛋白磷酸化表达量。将FAK反义寡核苷酸经脂质体转染VSMC,观察FAK mRNA及蛋白磷酸化、细胞迁移和黏附的变化。结果与对照组比较,PDGF组明显诱导细胞迁移和黏附,上调FAK mRNA的表达,提高FAK蛋白和磷酸化FAK表达,差异有统计学意义(P<0.05),与PDGF组比较,FAK组和雷帕霉素组细胞迁移、黏附能力减弱,FAK mRNA、FAK蛋白和磷酸化FAK表达水平明显降低,差异有统计学意义(P<0.05)。结论 PDGF诱导细胞迁移和黏附可能是FAK介导的,雷帕霉素可能是通过抑制FAK蛋白和磷酸化FAK来抑制VSMC的迁移和黏附。Objective To investigate the effect of focal adhesion kinase（FAK） on rapamycin（Rapa） inhibition of migration and adhesion of vascular smooth muscle eells（VSMC） induced by platelet derived growth factor（PDGF）. Methods VSMC were divided into four groups：the control group, PDGF group, Rapa ＋PDGF group （Rapa group） and FAK antisense oligodeoxynueleotides （FAKAODNS） ＋PDGF group（FAK group）. PDGF was used to stimulate the migration and adhesion of VSMC. The adhering cells and migrating cells were enumerated by light microscope and Boyden Chamber, FAK rnRNA, FAK protein and phosphorylation were detected by RT-PCR, Western blot and immunoprecipitation. FAKAODNS was transfected into VSMC by cationic lipid. The effects of FAKAODNS and Rapa on FAK mRNA and protein phosphorylation, VSMC migration and adhesion were investigated. Results Compare with the control group, PDGF could obviously induce VSMC adhesion and migration, up-regulate FAK mRNA expression, increase FAK protein and phosphorylated FAK expression （P 〈 0.05）. In FAK group and Rapa group, VSMC migration and adhesion were attenuated, expression of FAK mRNA, FAK protein and phosphorylated FAK decreased significantly （P 〈 0.05）. Conclusions Migration and adhesion of VSMC induced by PDGF may be mediated by FAK,Rapa may suppress the migration and adhesion of VSMC through inhibition of FAK protein and phosphorylated FAK.

关 键 词：黏着斑激酶1 西罗莫司 血小板源性生长因子 肌 平滑 血管 寡核苷酸类 反义 细胞黏附 

分 类 号：R965[医药卫生—药理学]