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心肌样细胞与内皮祖细胞联合移植治疗兔心肌梗死心室重构的实验研究

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

期数:
2010年第3期
页码:
322-326
栏目:
基础研究
出版日期:
2010-04-06

文章信息/Info

Title:
Effect of combined cell transplantation of cardiomyocyte-like cells and endothelial progenitor cells on ventricular remodeling after acute myocardial infarction in rabbits
作者:
郑翔; 方志成; 刘伯毅; 黄云飞; 陈黎
湖北郧阳医学院附属太和医院ICU，湖北 十堰 442000
Author(s):
ZHENG Xiang;  FANG Zhi-cheng;  LIU Bo-yi;  HUANG Yun-fei;  CHEN Li
Department of ICU, Taihe Hospital, Yunyang University of Medicine, Shiyan 442000, Hubei, China
关键词:
骨髓间充质干细胞; 心肌样细胞; 内皮祖细胞; 细胞移植
Keywords:
mensenchymal stem cell;  cardiomydium cell;  endothelial progenitor cell;  cell
分类号:
R457.7
DOI:
-
文献标识码:
A
摘要:
目的: 评价心肌样细胞和内皮祖细胞联合移植对心肌梗死心室重构疗效是否优于单一细胞移植。方法: 体外骨髓穿刺提取、分离、培养骨髓间充质干细胞（mensenchymal stem cell,MSC），向心肌样细胞和内皮祖细胞定向诱导分化。心肌梗死模型制作成功2周后，随机分为对照组、心肌样细胞组、内皮祖细胞组、联合移植组，沿梗死周边区域注射100 μl培养基或心肌样细胞、内皮祖细胞、二者混合悬液各3×1012/L。移植前、后4周心脏超声检查，TTC染色评价梗死面积、氯胺T氧化法测定血浆、心肌组织羟脯氨酸（hydroxyproline，Hyp）含量及基质金属蛋白酶含量。结果: ①诱导后MSC呈现心肌样细胞超微结构特征，PCR检查表达β-肌球蛋白重链（β-myosin heavy main，β-MHC）、受磷蛋白（phospholamban,PLB）、心房钠尿肽（atrial natriuretic polypeptide,ANP）；内皮祖细胞呈“铺路石”外观，高表达CD133并随时间推移表达逐步下降。②与对照组相比，细胞移植能改善心功能（P<0.05)，降低梗死面积（P<0.05），血浆羟脯氨酸含量（P<0.05），心肌组织羟脯氨酸（P<0.05）基质金属蛋白酶（MMP-9）含量(P<0.05)，联合细胞移植组效果更好。结论: MSC可定向分化心肌样细胞和内皮祖细胞，两者联合移植能够更好降低梗死面积、改善心室重构，效果优于单一细胞移植。
Abstract:
AIM: To compare the effect of combined cell transplantation of cardiomyocyte-like cells and endothelial progenitor cells with that of single-cell transplantation on the ventricular remodeling of rabbit acute myocardial infarction model. METHODS: Isolated mesenchymal stem cells (MSC) from the bone marrow of 24 rabbits were differentiated into cardiomyocyte-like cells and endothelial progenitor cells. Cell transplantation was performed 2 weeks after the establishment of acute myocardial infarction models. Rabbits were divided into control group, cardiomyocyte-like cell transplantation group, endothelial progenitor cell transplantation group and combined cell transplantation group. One hundred μl DMEM or cell suspension was injected into the border area of the myocardial infarction. Heart functions were assessed by transthoracic echocardiograph (TTC) before and 4 weeks after cell transplantation. Areas of infarction and constitution hydroxyproline were examined by TTC and chloraseptine oxidation method. RESULTS: MSCs induced by 5-AZA (5-azacytidine, 5-aza) and endothelial cell growth supplement (ECGS) from bovine pituitary were characterized by cardiomyocyte-like cells and endothelial progenitor cells. RT-PCR showed that the differentiated cells expressed atrial natriuretic polypeptide, phospholamban and myosin heavy chain mRNA. Endothelial progenitor cells showed a cobblestone shape when observed with phase-contrast microscope. CD133 significantly expressed and gradually descended in endothelial progenitor cells. Compared with control group, both combined cell transplantation and single cell transplantation ameliorated cardiac function and decreased the areas of infarction, plasma hydroxyproline or constitution hydroxyproline and MMP-9 (all P<0.05), but combined cell transplantation achieved better results than single cell transplantation. CONCLUSION: Our results show that MSCs can be differentiated into cardiomyocyte cells and endothelial progenitor cells in vitro. The effect of combined cell transplantation on ventricular remodeling of rabbit acute myocardial infarction model is superior to that of single cell transplantation.

参考文献/References

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备注/Memo

备注/Memo:
收稿日期：2009-04-24.基金项目：湖北省教育厅重点项目（B200624010） 通讯作者：方志成，副主任医师，主要从事干细胞移植研究Email:fang5280@yahoo.com.cn 作者简介：郑翔，副主任医师，硕士生Email:syzx481@163.com
更新日期/Last Update: 2010-04-09