BMP-2预诱导骨髓间充质干细胞移植治疗大鼠再灌注后心肌梗死的实验研究  

Effect of transplantation of BMP-2-induced bone marrow mesenchymal stem cells on myocardial infarction of rats after reperfusion

作　　者：苗忠澄 高航[1] 

机构地区：[1]辽宁医学院附属第一医院心内科,辽宁锦州121001

出　　处：《解放军医学杂志》2014年第8期601-608,共8页Medical Journal of Chinese People's Liberation Army

基　　金：辽宁省自然科学基金(201202130)~~

摘　　要：目的观察骨形态发生蛋白-2(BMP-2)能否诱导骨髓间充质干细胞(BMSCs)分化为类心肌细胞,以及BMSCs移植治疗大鼠急性心肌梗死(AMI)再灌注心肌的作用及对大鼠心功能的影响。方法体外分离大鼠BMSCs并进行培养扩增、鉴定,BMP-2预诱导后鉴定其心肌特有蛋白的表达。体外DAPI标记BMSCs。SD大鼠冠状动脉前降支(LAD)结扎100min后开通复流,建立AMI后再灌注模型。造模成功大鼠随机分为3组:对照组(在梗死周边区用微量注射器分4点注射培养基)、干细胞组(于相同部位注射等量未经诱导的BMSCs)、预诱导组(相同部位注射等量诱导后的BMSCs)。术后4h、4d及4周荧光显微镜观察被移植细胞是否存活于心肌组织中。术后4周采用彩色多普勒超声心动图检测心功能变化,并采用Masson、HE染色观察实验大鼠的心肌组织形态变化。结果经BMP-2预诱导后,部分BMSCs形态发生变化,表达心肌细胞特有蛋白缝隙连接蛋白43(Cx43)、肌钙蛋白T(cTnT)。术后4h、4d及4周时,荧光显微镜观察显示预诱导组及干细胞组均有DAPI标记的BMSCs存活在梗死心肌组织中。与对照组相比,干细胞组及预诱导组左室射血分数(LVEF)及左心室缩短分数(FS)均明显提高(P<0.01),预诱导组更为明显。4周后,HE染色显示,对照组梗死区炎性细胞浸润最严重,伴有严重纤维化,梗死呈连续性大片状分布,干细胞组梗死区炎性细胞浸润及纤维化较轻,预诱导组梗死区炎性细胞浸润及纤维化程度最轻。Masson染色可见干细胞移植部位出现胶原纤维,且随时间延长无明显变化,在胶原纤维中可见移植的细胞存活。结论 BMSCs经BMP-2诱导可向类心肌细胞分化,对AMI再灌注损伤有一定治疗作用;经BMP-2预诱导的BMSCs较未诱导的BMSCs治疗效果理想。Objective To observe whether the bone morphogenetic protein-2 （BMP-2） could induce bone marrow mesenchymal stem cells （BMSCs） to differentiate into cardiomyocyte-like cells, and investigate its effect on cardiac function in rats. Methods IBMSCs were isolated, cultured, amplified and identified in uitro. The expression of specific cardiac proteins was identified after pre-induction of BMP-2, and BMSCs were marked in vitro by DAPI. In order to reproduce the reperfusion after acute myocardial infarction （AMI） model, the left anterior descending （LAD） coronary artery was interrupted for 100min and then freed in SD rat. The rats with successful manipulation were randomly assigned into 3 groups, i.e. control group： the culture medium was injected with micro syringe at 4 points around infarcted zone; stem cell group： non-induced BMSCs were injected with the same amount and at the same sites as in control group; and pre-induction group： induced BMSCs were injected with the same amount and at the same sites as in control group. The surviving state of transplanted cells in myocardial tissue was observed with fluorescence microscopy at 4 hours, 4 days and 4 weeks postoperatively. Cardiac function was evaluated by employing color Doppler echocardiography, and the morphology of rat myocardial tissue was observed at 4 weeks after operation by HE and Masson staining, and the therapeutic effect was compared among 3 groups. Results After the BMP-2 pre-induction, changes in morphology were found in a part of BMSCs, and the specific cardiomyocyte protein connexin 43 （Cx43） and cardiac troponin T （cTnT） were expressed. Fluorescence microscopy revealed that DAPI-labeled BMSCs survived in myocardial infarction in pre-induction group and stem cell group 4 hours, 4 days and 4 weeks postoperatively. Compared with the control group, the left ventricular ejection fraction （LVEF） and left ventricular fractional shortening （FS） significantly increased in stem cell group and, especially, in pre- induction

关 键 词：心肌梗死 再灌注损伤 间质干细胞 骨形态发生蛋白质类 干细胞移植 

分 类 号：R542.22[医药卫生—心血管疾病]