ORAI1在损伤导致大鼠颈动脉新生内膜形成中的作用及其与钠钙交换体1的关系  

Role of ORAI1 in the balloon injury-induced rat carotid artery neointimal formation and the relationship between ORAI1 and sodium and calcium exchange 1

作　　者：黄世亮[1] 杨丽霞[1] 杨淑娟[1] 郭瑞威[1] 

机构地区：[1]成都军区昆明总医院心血管内科昆明医科大学,650032

出　　处：《中华心血管病杂志》2016年第11期968-972,共5页Chinese Journal of Cardiology

基　　金：国家自然科学基金（81370383）

摘　　要：目的 利用大鼠颈动脉球囊损伤模型探讨ORAI1在血管平滑肌细胞增生形成新生内膜中的作用及其与钠钙交换体（NCX）1的关系.方法 根据是否建立颈动脉球囊损伤模型和观察时间,将Sprague Dawley大鼠分为假手术组、损伤后7d组和损伤后14 d组,每组大鼠3只;根据转染的病毒液,将Sprague Dawley大鼠分为阴性对照组（建立颈动脉球囊损伤模型后,转染阴性对照慢病毒质粒）和siORAI1组（建立颈动脉球囊损伤模型后,转染沉默ORAI1的慢病毒质粒）,每组3只大鼠.采用HE染色评价颈动脉内膜的增生情况;采用Real-time PCR检测血管壁ORAI1的mRNA表达水平;采用Western blot检测血管壁核增殖核抗原（PCNA）、ORAI1和NCX1的蛋白表达水平.结果 （1）损伤后7d组和损伤后14 d组的颈动脉内膜厚度均大于假手术组[（0.54 ±0.11）μ m2和（0.89±0.12） μm2比（0.11 ±0.08） μm2,P均＜0.05],PCNA的蛋白相对表达水平均高于假手术组（1.43±0.16和1.95±0.16比1,P均＜0.05）,ORAI1的mRNA表达水平均高于假手术组（1.39 ±0.14和1.78 ±0.21比0.56±0.09,P均＜0.05）,ORAI1的蛋白相对表达水平均高于假手术组（1.42 ±0.19和1.78±0.22比1,P均＜0.05）.（2）球囊损伤后14 d,siORAI1组的ORAI1蛋白相对表达水平低于阴性对照组（0.21 ±0.16比1,P＜0.05）,颈动脉内膜厚度小于阴性对照组[（0.19±0.14） μm2比（0.91 ±0.23） μm2,P＜ 0.05],NCX1蛋白相对表达水平低于阴性对照组（0.53 ±0.13比1,P＜0.05）.结论 ORAI1可能是调控血管新生内膜形成和细胞增殖的一个关键因素;ORAI1基因沉默可能会引起NCX1表达水平的降低.Objective To investigate the role of ORAI1 in the balloon injury-induced rat carotid artery neointimal formation and the relationship between ORAI1 and sodium and calcium exchanger（NCX） 1.Methods According to condition of carotid artery balloon injury and observation time,Sprague Dawley rats were divided into 3 groups including sham group,7 days injury group,and 14 day injury group （n =3 each）.According to virus of transfection,Sprague Dawley rats were divided into 2 groups including negative control group （negative lentiviruses particles transfection after establishment of carotid artery balloon injury model,n =3） and siORAI1 group（siORAI1 transfection after operation,n =3）.Carotid artery neointimal formation was analyzed after HE staining.The ORAI mRNA expression level was detected using real-time PCR.The proliferating cell nuclear antigen （PCNA）,ORAI and NCX1 protein expression level were measured by Western blot.Results （1） Carotid artery intima was significantly thicker in 7 days and 14 days injury group than in sham group（（0.54 ±0.11） μm2 and （0.89 ±0.12） μm2 vs.（0.11 ±0.08） μm2,both P 〈0.05）.Relative expression level of PCNA protein was significantly higher in 7 days and 14 days injury group than in sham group（1.43 ±0.16 and 1.95 ±0.16 vs.1,both P 〈0.05）.Relative mRNA expression level of ORAI1 （1.39 ± 0.14 and 1.78 ± 0.21 vs.0.56 ± 0.09,both P 〈 0.05） and protein expression level （1.42 ± 0.19 and 1.78 ±0.22 vs.1,both P 〈0.05） were significantly higher in 7 days and 14 days injury group than in sham group.（2） After 14 days,relative expression level of ORAI1 protein was significantly lower in siORAI1 group than in negative control group （0.21 ±0.16 比 1,P 〈0.05）.Carotid artery intima thickness was significantly reduced in siORAI1 group compared to negative control group（（0.19 ± 0.14）μm2 vs.（0.91 ±0.23） μm2,P〈0.05）.Relative expression level of NCX1 protein was also significantly lower in siORAI1 group than i

关 键 词：肌细胞 平滑肌 血管内膜 增生 

分 类 号：R543[医药卫生—心血管疾病]