上海交通大学学报(医学版)

• 论著(基础研究) • 上一篇    下一篇

血管紧张素转化酶2基因转染对动脉硬化血管内皮细胞的保护作用

陈 旭1,朱 莉1,郝青青1,于庆涛1,李树英1,张月辉1,2,董 波1   

  1. 1.山东大学附属省立医院心内科, 济南 250021; 2.南方医科大学附属深圳宝安医院重症医学科, 深圳 518101
  • 出版日期:2014-04-28 发布日期:2014-05-13
  • 通讯作者: 张月辉, 电子信箱: dbsh2004@163.com; 董 波, 电子信箱: dongbo1@medmail.com.cn。
  • 作者简介:陈 旭(1988—), 女, 硕士生; 电子信箱: chenxusophy@sina.com。
  • 基金资助:

    国家自然科学基金(81170207)

Protective effect of transfection of angiotensin converting enzyme 2 gene on atherosclerotic vascular endothelial cells

CHEN Xu1, ZHU Li1, HAO Qing-qing1, YU Qing-tao1, LI Shu-ying1, ZHANG Yue-hui1,2, DONG Bo1   

  1. 1.Department of Cardiology, Shandong Provincial Hospital, Shandong University, Jinan 250021,China; 2.Department of Critical Care Medicine, the Affiliated Baoan Hospital of Southern Medical University, Shenzhen 518101,China
  • Online:2014-04-28 Published:2014-05-13
  • Supported by:

    National Natural Science Foundation of China, 81170207

摘要:

目的 通过观察血管紧张素转化酶2(ACE2)基因转染对动脉粥样硬化模型兔血管内皮细胞的作用,阐明ACE2基因转染对内皮细胞的保护作用及机制。方法 采用球囊损伤内皮细胞及高脂饲养建立兔动脉粥样硬化模型,16只新西兰大白兔随机分为Ad-ACE2组(转染携带ACE2基因的重组腺病毒)和Ad-EGFP组(转染空载重组腺病毒),每组8只。Ad-ACE2组和Ad-EGFP组分别经腹主动脉注射ACE2腺病毒(2.5×109 pfu/mL)和Ad-EGFP。1个月后处死动物取腹主动脉制备标本,Western blotting法检测动脉粥样硬化斑块中ACE2蛋白表达,免疫组织化学法检测细胞间黏附分子-1 (ICAM-1)和P选择素(P-selectin)蛋白的表达,油红O染色进行脂质半定量分析。结果 Ad-ACE2组斑块中ACE2蛋白表达显著高于Ad-ACE2组(P<0.05)。Ad-ACE2组斑块内ICAM-1和P-selectin的阳性表达率均明显低于Ad-EGFP组[(16.95±3.09)%和(24.81±4.78)%;(16.22±2.45)%和(23.46±3.28)%],组间比较差异均有统计学意义(P<0.01)。Ad-ACE2组斑块内脂质含量为(41.77±3.17)%,明显低于Ad-EGFP组的(59.55±1.53)%,组间比较差异有统计学意义(P<0.05)。结论 ACE2基因转染通过下调ICAM-1和P-selectin蛋白的表达,保护兔动脉内皮细胞,抑制动脉粥样硬化的进展。

关键词: 血管紧张素转化酶2, 基因转染, 动脉粥样硬化, 内皮损伤

Abstract:

Objective To explain the protection effect and mechanism of gene transfection of angiotensin converting enzyme 2 (ACE2) towards endothelial cells by observing the effects of gene transfection of ACE2 on vascular endothelial cells of model rabbits with atherosclerosis. Methods The atherosclerosis model of rabbit was established by balloon injuries of endothelial cells and atherogenic diet. Sixteen New Zealand rabbits were randomly divided into the Ad-ACE2 group (transfected by the recombinant adenovirus carrying the ACE2 gene) and Ad-EGFP group (transfected by the recombinant adenovirus without carrying the ACE2 gene). Each group consisted of 8 rabbits. The rabbits of two groups received abdominal aortic injections of ACE2 recombinant adenovirus (2.5×109 pfu/mL) and Ad-EGFP, respectively. After one month, all rabbits were sacrificed and specimens of plaques from abdominal aorta segments were prepared. The protein expressions of ACE2 in atherosclerotic plaques were detected by the Western blotting. The expressions of intercellular adhesion molecule-1 (ICAM-1) and P-selectin protein were evaluated by the immunohistochemical staining. The semi-quantitative analysis was carried out by the oil red O staining. Results The results showed that the expressions of ACE2 protein of the Ad-ACE2 group were significantly higher than that of the Ad-EGFP group (P<0.05). The positive expression rates of ICAM-1 and P-selectin protein in plaques of the Ad-ACE2 group were significantly lower than those of the Ad-EGFP group [(16.95±3.09)% vs (24.81±4.78)% and (16.22±2.45)% vs (23.46±3.28)%, respectively]. The differences between two groups were statistically significant (P<0.01). The lipid contents in plaques of the Ad-ACE2 group (41.77±3.17)% were significantly lower than those of the Ad-EGFP group (59.55±1.53)%. The differences between two groups were statistically significant (P<0.05). Conclusion ACE2 gene transfection can protect the endothelial cells of rabbits and inhibit the development of atherosclerosis by down-regulating the expressions of ICAM-1 and P-selectin protein.

Key words: angiotensin converting enzyme 2, gene transfection, atherosclerosis, endothelial injury