同种异体骨髓间充质干细胞在大鼠体内心肌缺血微环境中Nav1.5的表达  

Expression of Nav1.5 on allogeneic mesenchymal stem cells in the microenvironment of rat myocardial ischemia in vivo

作　　者：张静[1] 王亭忠[2] 魏峰[2] 倪雅娟[3] 马爱群[2] 

机构地区：[1]新疆医科大学第四附属医院心脏中心,乌鲁木齐830000 [2]西安交通大学医学院第一附属医院心内科 [3]西安交通大学医学院第二附属医院心内科

出　　处：《山西医科大学学报》2014年第10期907-910,1001,共5页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(30800455)

摘　　要：目的观察大鼠同种异体骨髓间充质干细胞(mesenchymal stem cells,MSCs)在体内心肌缺血微环境中向心肌细胞分化过程中Nav1.5的分化表达。方法采用左冠状动脉前降支(left anterior descending coronary artery,LAD)结扎术建立大鼠急性心肌梗死(myocardial infarction,MI)模型(n=50),将绿色荧光蛋白(green fluorescent protein,GFP)转染标记MSCs,并贴壁培养,用心外膜下注射的方法将GFP标记的MSCs移植到心肌梗死周边区;通过免疫荧光染色观察移植后3,5,7 d移植的MSCs上Nav1.5蛋白的表达;应用激光捕获显微切割技术分离移植后3,5,7 d心肌组织中GFP标记的MSCs群,应用realtime PCR测定移植后3,5,7 d Nav1.5及未移植MSCs mRNA的相对表达量;对移植后9 d的移植区域行TUNEL凋亡染色。结果免疫荧光染色观察移植的MSCs于移植后3,5,7 d均不表达Nav1.5;real-time PCR测定显示Nav1.5相对表达量在移植后3,5,7 d之间比较及分别与未移植的MSCs相比均无明显变化(P>0.05);移植的MSCs于移植后9 d在心肌组织中几乎观察不到,TUNEL凋亡染色显示移植的MSCs发生了凋亡。结论在大鼠体内心肌缺血微环境中移植的同种异体MSCs不表达Nav1.5且不能长期存活。Objective To observe the expression of Nav l. 5 on allogeneie mesenchymal stem cells （MSCs） in microenvironment of rat myocardial ischemia in vivo. Methods The myocardial infarction（MI） rat models（n =-50） were established by ligating the left ante- rior descending coronary artery（LAD）. The MSCs were labeled by green fluorescent protein（GFP） and then cultured using direct ad- herent culture methods. GFP-labeled MSCs were injected into the surrounding area of the epicardium myocardial infarction. The protein expression of Navl. 5 in the transplanted MSCs was observed by immunofluorescence staining at day 3, 5 and 7, and the GFP-labeled MSCs were acquired by laser capture microdissection and the relative mRNA expression of Navl. 5 and untransplanted MSCs was meas- ured by real-time PCR. The transplanted areas were observed by TUNEL staining at day 9. Results The transplanted MSCs did not express Navl. 5 at day 3, 5, 7 after transplantation. Compared with non-transplanted MSCs, the relative expression of Navl. 5 was not significantly different in transplanted MSCs at day 3, 5, 7 or among each other by real-time PCR（ P 〉 0.05 ）. The transplanted MSCs were hardly observed in myocardial tissues at day 9. TUNEL staining showed that MSCs were apoptotic. Conclusion The transplan- ted allogeneic MSCs could not express Navl. 5, and could not survive for a long time effectively.

关 键 词：心肌梗死 骨髓间充质干细胞 移植 NAV1.5 

分 类 号：R541[医药卫生—心血管疾病]