[1]赵然尊,龙仙萍,刘志江,等.转染hRAMP1的间充质干细胞对心肌梗死后局部血管再生的作用[J].第三军医大学学报,2012,34(15):1496-1500.
 Zhao Ranzun,Long Xianping,Liu Zhijiang,et al.MSCs with hRAMP1 overexpression improve revascularization in a rabbit model of myocardial infarction[J].J Third Mil Med Univ,2012,34(15):1496-1500.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
34卷
期数:
2012年第15期
页码:
1496-1500
栏目:
论著
出版日期:
2012-08-15

文章信息/Info

Title:
MSCs with hRAMP1 overexpression improve revascularization in a rabbit model of myocardial infarction
作者:
赵然尊龙仙萍刘志江王冬梅石蓓
遵义医学院第一附属医院心血管内科
Author(s):
Zhao Ranzun Long Xianping Liu Zhijiang Wang Dongmei Shi Bei
Department of Cardiology, First Affiliated Hospital, Zunyi Medical College, Zunyi, Guizhou Province, 563000, China
关键词:
受体活性修饰蛋白-1间充质干细胞心肌梗死再血管化细胞因子
Keywords:
receptor activity modified protein-1 mesenchymal stem cells myocardial Infarction (MI) revascularization cytokine factor
分类号:
R364.33; R394.1; R542.22
文献标志码:
A
摘要:
目的      探讨腺病毒介导hRAMP1基因转染间充质干细胞(mesenchymal stem cells,MSCs)移植对心肌梗死后新生血管生成及其可能机制。      方法      密度梯度离心并贴壁培养获得兔骨髓MSCs,并分别转染pAd2-hRAMP1或pAd2-EGFP 后给予CGRP刺激,ELISA法测定细胞培养上清液中VEGF和HGF水平。建立心肌梗死再灌注兔模型,采用随机数字表法分为MSChRAMP1组(pAd2-hRAMP1转染MSCs移植)、MSCnull组(pAd2-EGFP转染MSCs移植)组和对照组(等量生理盐水注射)(n=10)。2周时Western blot检测心肌梗死组织促血管生长因子VEGF和HGF蛋白表达;4周时TTC染色测定心肌梗死面积比;抗CD31免疫组化染色检测心肌梗死区及梗死周边区新生毛细血管密度。      结果       CGRP诱导72 h后MSChRAMP1培养上清液中VEGF和HGF水平显著高于MSCnull组[VEGF:(1 859.4±267.4) vs (1 344.4±137.2);(1 052.2±239.3) vs (683.8±150.5)pg/ml,P<0.05]。细胞移植后4周TTC染色检测显示MSChRAMP1组心肌梗死面积显著降低对照组和MSCnull组[(10.1±2.9)% vs(30.6±2.7)%和(22.5±3.2)%,P<0.05];抗CD31免疫组化染色显示,MSChRAMP1组的梗死区和梗死交界区新生毛细血管计数明显高于对照组和MSCnull组(P<0.05);Western blot检测显示细胞移植后2周MSChRAMP1组心肌梗死区促血管生长因子VEGF和HGF蛋白表达显著高于对照组和MSCnull组(P<0.05)。      结论        MSChRAMP1能通过提高梗死区心肌组织中促血管生长因子VEGF和HGF的表达,促进心肌梗死区和梗死交界区新生毛细血管生成,降低心肌梗死面积。
Abstract:
Objective      To determine the effect of human receptor activity modified protein 1 (hRAMP1)-overexpressing mesenchymal stem cells (MSCs) by adenovirus vector on infarcted related revascularization and its potential mechanism in a rabbit model of myocardial infarction.       Methods        Thirty 4- to 5-month-old health male rabbits were developed to myocardial infarction (MI) and reperfusion models by ligating left ventricular branch of coronary artery for 60 min, and then divided into MSChRAMP1 group (n=10), MSCnull group (n=10) and control group (n=10) according to whether they were received intravenously injection of MSCs transfected with adenovirus vector encoding hRAMP1 gene and enhanced green fluorescent protein (EGFP), of MSCs transfected with adenovirus vector encoding only EGFP without hRAMP1 gene or of equally volume of phosphate buffered saline (PBS).The protein expression of vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) in infracted myocardium were measured by Western blotting in 2 weeks following MI. MI was evaluated by TTC staining for infracted area, and capillary density in infarcted region and its border region were tested by anti-CD31 immunohistochemistry staining. VEGF and HGF levels in the supernatant were quantified by ELISA assay in 24 and 72 h after induction of CGRP in cultured MSCs.       Results       The levels of VEGF and HGF in the cultured supernatant of MSChRAMP1 group were significantly higher than in the supernatant of MSCnull group when they were induced by CGRP for 72 h. Furthermore, there was a significant decrease in the area ratio of MI in MSChRAMP1 group than in MSCnull and control group by TTC staining. Also, compared with MSCnull and control groups, capillary density indicated by CD31-positive cells in the infarcted and border region was statistically augmented in MSChRAMP1 group by immunohistochemistry assay. The protein expression of VEGF and HGF detected by Western blotting were much higher in MSChRAMP1 group compared with MSCnull and control group at 2 weeks following MSCs delivery.       Conclusion        Gene modified MSCs with hRAMP1 overexpression have the powerful potential to increase capillary density and improve revascularization through more secretion of VEGF and HGF in infarcted and border region.

参考文献/References:

赵然尊, 龙仙萍, 刘志江, 等. 转染hRAMP1的间充质干细胞对心肌梗死后局部血管再生的作用[J].第三军医大学学报,2012,34(15):1496-1500.

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更新日期/Last Update: 2012-07-26