降钙素基因相关肽调节平滑肌细胞表型改变对动脉损伤后新生内膜增生的作用  

Effect of calcitonin gene-related peptide on vascular smooth muscle cells phenotypic switch and neointima formation after artery injury

作　　者：赵然尊[1] 龙仙萍[1] 陈攀科[1] 崔璨[1] 盛瑾[1] 王冬梅[1] 石蓓[1] 

机构地区：[1]遵义医学院第一附属医院心血管内科,贵州遵义563003

出　　处：《中华高血压杂志》2014年第11期1036-1042,共7页Chinese Journal of Hypertension

基　　金：国家自然科学基金(NSFC 30860100);贵州省国际合作项目[黔科合外G字(2010)0732]

摘　　要：目的通过降钙素基因相关肽(CGRP)干预体外培养的血管平滑肌细胞(VSMC)和体内颈动脉损伤大鼠模型,探讨其对VSMC表型改变和动脉损伤后新生内膜增殖的影响。方法取大鼠胸主动脉,以组织块贴壁培养法获得VSMC,给予血小板源生长因子(PDGF,终浓度为2.0×10-8 mol/L)和(或)CGRP(终浓度为3.5×10-7 mol/L)处理,利用Western blot法检测细胞表型改变,噻唑蓝法和流式细胞仪检测细胞增殖,细胞划痕实验检测细胞迁移能力;同时建立颈动脉损伤大鼠模型,并分为对照组和CGRP组,通过HE染色观察新生内膜形成情况,Western blot法检测损伤动脉中VSMC表型改变和信号转导通路细胞外信号调节激酶(ERK)激活情况。结果PDGF明显诱导体外培养的VSMC表型改变;而CGRP处理则显著抑制PDGF诱导的细胞表型转换,降低细胞增殖[48h:CGRP组(0.20±0.03)×105比PDGF组(0.67±0.08)×105,P<0.05;72h:CGRP组(0.17±0.04)×105比PDGF组(1.42±0.11)×105比对照组(0.45±0.05)×105,P<0.05]和迁移能力[24h:CGRP组(2.12±0.21)%比PDGF组(5.04±0.52)%比对照组(4.87±0.44)%,P<0.05;48h:CGRP组(2.84±0.43)%比PDGF组(5.78±0.74)%比对照组(5.50±0.37)%,P<0.05];CGRP降低新生内膜形成,伴随着损伤动脉中VSMC合成表型标志蛋白骨桥蛋白表达减少,并抑制细胞外信号激活途径ERK的激活。结论 CGRP可能通过抑制ERK1/2信号途径激活而调节VSMC表型,抑制细胞增殖和迁移,从而降低动脉损伤后新生内膜形成。Objective To investigate the effects of calcitonin gene-related peptide（CGRP）on vascular smooth muscle cells（VSMC）phenotypic switch and neointima formation after artery injury through CGRP-treated cultured VSMC in vitro and balloon-injured rat carotid artery in vivo. Methods VSMC were obtained by aortic tissue adherent culture method and treated with platelet-derived growth factor（PDGF,2.0×10-8 mol/L）and/or CGRP（3.5×10-7 mol/L）. Phenotypic switch of VSMC was detected by Western blot,cell proliferation was determined by both MTT and FACS,and cell migration ability was assessed by scratch test. Furthermore,balloon-injured rat carotid artery was induced and the rats were divided into control group and CGRP group,in which neointima formation was observed by HE staining. VSMC phenotypic switch and extracellular signal-regulated kinase（ERK）activity in injured artery was detected by Western blot. Results In vitro cultured VSMC,PDGF induced obviously cell phenotypic changes,while CGRP treatment significantly suppressed cells phenotypic switch,reduced cell proliferation[48h：CGRP group（0.20±0.03）×105 vs PDGF group（0.67±0.08）×105,P〈0.05;72h：CGRP group（0.17±0.04）×105 vs PDGF group（1.42±0.11）×105 or control group（0.45±0.05）×105,P〈0.05]and migration ability[24h：CGRP group（2.12±0.21）% vs control group（4.87±0.44）% or PDGF group（5.04±0.52）%,P〈0.05;48h：CGRP group（2.84±0.43）% vs control group（5.50±0.37）% or PDGF group（5.78±0.74）%,P〈0.05）]induced by PDGF.In the rat model of carotid artery injury,CGRP significantly reduced neointima formation,accompanied with the decrease of osteopontin（OPN）expression（a synthetic phenotypic marker protein of VSMC）and the inhibition of ERK activation. Conclusion CGRP may regulate VSMC phenotypic switch and inhibit cell proliferation and migration through the inhibition of ERK1/2signal pathway,and therefore reduce neointima formation after arterial injury.

关 键 词：血管平滑肌细胞 降钙素基因相关肽 表型改变 

分 类 号：R543.5[医药卫生—心血管疾病]