受体活性修饰蛋白1基因载体构建及其对大鼠血管平滑肌细胞增殖的影响  

Construction of Receptor Activity Modifying Protein Gene Vector and Effect on Proliferation of Rat VSMC

作　　者：郑元斌[1] 秦旭平[1] 孙飞[1] 唐江琼[1] 廖端芳[1] 

机构地区：[1]南华大学药物药理研究所,湖南衡阳421001

出　　处：《南华大学学报（医学版）》2010年第1期1-5,共5页Journal of Nanhua University(Medical Edition)

基　　金：国家自然科学基金(30572192);湖南省自然科学基金(05JJ30042)

摘　　要：目的探讨受体活性修饰蛋白1(RAMP1)表达载体的构建及其高表达RAMP1对血管平滑肌细胞增殖的影响。方法通过构建RAMP1大鼠基因开放阅读框的真核表达载体,利用脂质体将其转入原代培养的大鼠主动脉平滑肌细胞,G418筛选稳定表达细胞集落。逆转录-聚合酶链反应和蛋白印迹法检测RAMP1的表达量;MTT法测定细胞的增殖或生存率,并计算细胞倍增时间。结果成功构建RAMP1基因真核表达载体,脂质体能将RAMP1表达载体稳定转入血管平滑肌细胞,RAMP1高表达能明显增强降钙素基因相关肽抑制血管紧张素Ⅱ诱导的血管平滑肌细胞生存率及明显延长其倍增时间。结论RAMP1高表达能增强CGRP抑制血管紧张素Ⅱ诱导的血管平滑肌细胞增殖。Objective To explore construction of receptor activity modifying protein（RAMP1）,a eukaryotic expression vector of Rat RAMP1 ORF,and study the effect of RAMP1 hyperexpression on proliferation of vascular smooth muscle cell（VSMC）. Methods Lipofectin was utilized to transfer the RAMP1 ORF vector into rat thoracic aortas VSMC,stable transfection cell colony of express RAMP1 was obtained by G418 screening.RT-PCR and Western-blotting were used to detect the expression quantity of RAMP1.VSMC viability or proliferation was determined by MTT and doubling time. Results Vector of RAMP1 was successfully constructed,and lipofectin can stably transfect the vector of RAMP1 to VSMC,and overexpression of RAMP1 significantly decrease the ratio of viability and doubling time of VSMC induced by angiotensin Ⅱ. Conclusions RAMP1 overexpression can significantly enhance the inhibitory effect of CGRP on the proliferation of VSMC induced by Angiotensin Ⅱ.

关 键 词：受体活性修饰蛋白1 降钙素基因相关肽 血管平滑肌细胞 增殖 

分 类 号：R972[医药卫生—药品]