生胃酮对大鼠颈动脉损伤后新生内膜过度增殖的影响  

Effect of Carbenoxolone on Neointimal Hyperplasia after Rat Carotid Balloon Injury

作　　者：宋明宝[1] 于学军[1] 崔馨[2] 陈剑飞[1] 卢来春[1] 赵刚[1] 于世勇[1] 董红梅[1] 黄岚[1] 

机构地区：[1]第三军医大学新桥医院全军心血管病研究所,重庆400037 [2]第三军医大学大坪医院,重庆400037

出　　处：《儿科药学杂志》2009年第5期10-13,共4页Journal of Pediatric Pharmacy

基　　金：国家自然科学基金;项目编号:30570765;30700889;重庆市自然科学基金;项目编号:CSTC;2005bb5304

摘　　要：目的:观察生胃酮对大鼠颈动脉损伤后新生内膜过度增殖的影响。方法:采用组织贴块法培养大鼠主动脉平滑肌细胞,Western blot检测生胃酮对大鼠主动脉平滑肌细胞缝隙连接蛋白43表达的影响。采用球囊损伤建立大鼠颈动脉损伤模型,对照组给予生理盐水2 mL、生胃酮组给予生胃酮3 mg/kg腹腔注射,1次/d,HE染色观察新生内膜形成情况,免疫荧光染色检测新生内膜中缝隙连接蛋白43的表达,伊文思蓝-DAPI双染色以评价新生内膜形成情况。结果:所培养的细胞抗SMα-actin免疫荧光染色阳性。50μmol/L生胃酮处理平滑肌细胞24 h后对缝隙连接蛋白43的表达无影响(0.85±0.06vs0.83±0.03,n=3,P>0.05)。球囊损伤大鼠颈动脉14 d后可见血管管腔狭窄、新生内膜增生明显。免疫荧光染色显示形成的新生内膜中缝隙连接蛋白43表达丰富。经生胃酮干预后新生内膜增生明显减轻,新生内膜细胞计数显著低于对照组(89±28.40vs236±15.04,n=5,P<0.01)。结论:缝隙连接在血管损伤后新生内膜形成过程中扮演重要角色,而缝隙连接阻断剂生胃酮能抑制新生内膜过度增生。Objective： To observe the effect of carbenoxolone on neointimal hyperplasia after rat carotid balloon injury. Methods： Rat aortic SMCs （RASMCs） were cultured by explanted rat aortic wall tissue and identified with cell inanunofluorescence staining for SM α-actin. The expression of Cx43 in RASMCs which were treated with carbenoxolone at a concentration of 50μmol/L for 24 h was detected with western blot. The model of vascular injury was established with rat carotid balloon injury. And animals were administrated with intrapefitoneal injections of carbenoxolone [3 mg/（ kg· d） ] in the carbenoxolone group or with saline （2 mL/d） in the control group for 2 weeks after carotid balloon injury. After 2 weeks, HE staining and DAPI-Evens blue double staining were applied to evaluate the neointimal formation of targeted vessels. And cell immunofluorescence staining was used to detect protein Cx43 expression on targeted vessels. Results： RASCMCs were cultured successfully with positive immunofluoreseence staining for SM α-actin. In this experiment, we found that carbenoxolone couldn＇t induce the expression of Cx43 protein in RASMCs （0.85 ± 0.06 vs 0. 83 ±0.03, n = 3, P 〉 0.05）. Two weeks after carotid balloon injury, carbenoxolone could significantly reduce the neointimal formation and the stenosis of blood vessel lumen. When nuclei number of neointima was used to evaluate the neointimal formation, result suggested that nuclei number of neointima in carbenoxolone group was significantly lower than that in the control group （ 89 ± 28.40 vs 236 ± 15.04, n = 5, P 〈 0.01 ）. The expression of Cx43 protein in neointima was abundant. Conclusions： Gap junctions play a key role in neointimal hyperplasia, and carbenoxolone, a special blocker of gap junction, can inhibit neointimal hyperplasia after rat carotid balloon injury.

关 键 词：新生内膜 血管 缝隙连接 生胃酮 大鼠 

分 类 号：R363[医药卫生—病理学]