中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (46): 8598-8602.doi: 10.3969/j.issn.1673-8225.2011.46.012

• 血管组织构建 vascular tissue construction • 上一篇    下一篇

腺病毒介导hRAMP1基因转染血管平滑肌细胞的增殖和凋亡

龙仙萍，赵然尊，石  蓓，崔  璨，盛  瑾，陈攀科   

  1. 遵义医学院第一附属医院心血管内科，贵州省遵义市  563003
  • 收稿日期:2011-02-18 修回日期:2011-04-24 出版日期:2011-11-12 发布日期:2011-11-12
  • 通讯作者: 石蓓，教授，硕士生导师，遵义医学院第一附属医院心血管内科，贵州省遵义市 563003 shibei2147@yahoo.com.cn
  • 作者简介:龙仙萍★，女，1980年生，贵州省遵义市人，苗族，2011年遵义医学院毕业，硕士，主治医师，主要从事冠心病介入治疗后再狭窄的基础与临床研究。 Longxianping1201@163.com
  • 基金资助:

    国家自然科学基金(NSFC30860100)：心肌梗死后RAMP1基因修饰的MSCs移植对血管成形术后再狭窄的作用研究”；贵州省国际合作项目(黔科合外G字[2010]0732)：RAMP-1基因修饰MSCs移植治疗心肌梗死的实验研究”。

Effect of adenovirus mediated human RAMP1 gene on proliferation and apoptosis of vascular smooth muscle cells

Long Xian-ping, Zhao Ran-zun, Shi Bei, Cui Can, Sheng Jin, Chen Pan-ke   

  1. Department of Cardiology, First Affiliated Hospital of Zunyi Medical College, Zunyi 563003, Guizhou Province, China
  • Received:2011-02-18 Revised:2011-04-24 Online:2011-11-12 Published:2011-11-12
  • Contact: Shi Bei, Professor, Master’s supervisor, Department of Cardiology, First Affiliated Hospital of Zunyi Medical College, Zunyi 563003, Guizhou Province, China shibei2147@yahoo.com.cn
  • About author:Long Xian-ping★, Master, Attending physician, Department of Cardiology, First Affiliated Hospital of Zunyi Medical College, Zunyi 563003, Guizhou Province, China longxianping1201@163.com
  • Supported by:

    the National Natural Science Foundation of China, No. NSFC30860100*; International Cooperation Program of Guizhou Province, No. qiankehewai G[2010]0732*.

PDF

341

被引次数

9

摘要：

背景：外源性受体活性修饰蛋白1基因转染可能对血管平滑肌细胞增殖调控具有重要的作用。
目的：探讨腺病毒载体介导人受体活性修饰蛋白1基因对体外培养的兔血管平滑肌细胞增殖和凋亡的影响。
方法：分离培养获得兔主动脉血管平滑肌细胞，分别以携带人受体活性修饰蛋白1基因的腺病毒和空腺病毒转染后，分成人受体活性修饰蛋白1组、空病毒组和对照组。
结果与结论：腺病毒转染细胞后随时间延长，报告基因绿色荧光蛋白表达逐渐增加，72 h表达最强，感染率达80%，96 h时仍有绿色荧光蛋白表达。人受体活性修饰蛋白1组的受体活性修饰蛋白1蛋白表达增加，血管平滑肌细胞凋亡率增加，细胞增殖明显抑制，与空病毒组和对照组比较差异有显著性意义(P < 0.05)。提示人受体活性修饰蛋白1基因感染血管平滑肌细胞后明显抑制血管平滑肌细胞增殖，促进细胞凋亡。

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, 凋亡

Abstract:

BACKGROUND: Hyperproliferation of vascular smooth muscle cells (VSMCs) is one of the most important mechanisms of atherosclerosis and restenosis. Receptor activity modifying protein-1 (RAMP1) is receptor subunit of calcitonin gene related peptide (CGRP) and decides the activity of receptor. Exogenous RAMP1 could play an important role in the regulation of VSMCs proliferation. 
OBJECTIVE: To investigate the effects of adenovirus mediated human RAMP1 (hRAMP1) gene on proliferation and apoptosis of VSMCs derived from rabbit thoracic aorta.
METHODS: VSMCs, isolated from rabbit thoracic aorta, were divided into three groups: hRAMP1 group, empty-adenovirus vector group and control group according to whether transferring Ad-EGFP-hRAMP1 or Ad-EGFP, and each group included four subgroups of 24 hours, 48 hours, 72 hours and 96 hours. RAMP1 protein was detected by immunocytochemistry, and the proliferation of VSMCs was determined by cell counting and MTT assay, and the apoptosis of VSMCs was determined by flow cytometry and TUNEL assay.
RESULTS AND CONCLUSION: VSMCs were successfully infected by Ad-EGFP-hRAMP1 or Ad-EGFP, and EGFP expression was increased, peaked at 72 hours and lasted for 96 hours. In the hRAMP1 group, the VSMCs showed hRAMP1 protein expression, and the activity of cell proliferation was significantly inhibited in the hRAMP1 group compared with empty adenovirus vector group and control group (P < 0.05). Also, the level of VSMCs apoptosis was significantly higher in the hRAMP1 group compared with other groups (P < 0.05). These results showed that the over-expression of hRAMP1 mediated by adenovirus vector plays an important role in inhibition of VSMCs proliferation and promotion of VSMCs apoptosis, which may be the therapeutic target of atherosclerosis and restenosis.

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