骨髓源性心肌干细胞移植治疗心肌梗死的实验研究  

Effects of marrow-derived cardiac stem cell transplantation after myocardial infarction in rats

作　　者：张贵焘[1] 谭玉珍[1] 王海杰[1] 敖红[1] 国海东[1] 黄国英[2] 

机构地区：[1]复旦大学上海医学院人体解剖与组织胚胎学系,200032 [2]复旦大学附属儿科医院心血管中心

出　　处：《中华心血管病杂志》2007年第10期940-944,共5页Chinese Journal of Cardiology

基　　金：国家自然科学基金(30470883)

摘　　要：目的研究骨髓源性心肌干细胞(MCSC)移植对心肌梗死的治疗作用。方法通过单细胞克隆培养技术从雄性 SD 大鼠骨髓间充质干细胞(MMSC)中筛选 MCSC。结扎雌性 SD 大鼠的左冠状动脉前降支,建立心肌梗死模型,1周后于梗死区边缘移植 MMSC 和 MCSC。移植后4周,用超声心动图检测心功能变化。取心肌组织作冷冻切片,用 HE 和 Masson 染色法显示瘢痕区的组织结构变化,通过免疫组织化学染色标记血管内皮牛长因子受体-1阳性(VEGFR-1^+)微血管,用图像分析系统测量瘢痕而积和微血管密度。利用原位荧光杂交标记含有 Y 染色体的 MCSC,并检测心肌特异性肌钙蛋白 T(cTnT)的表达。结果筛选的 MCSC 表达 c-kit,心肌早期转录因子 Nkx2.5呈低表达。细胞移植后4周,MCSC 移植组的左室短轴缩短分数(62.9%±2.2%)和左室射血分数(32.8%±1.1%)高于 MMSC 移植组(分别为55.7%±1.6%和28.2%±1.6%)和对照组(分别为42.4%±2.1%和23.6%±1.2%);MCSC 组心肌梗死面积比率(8.7%±0.7%)低于 MMSC 组(12.0%±1.1%)和对照组(16.8%±0.9%)。含有 Y 染色体的 MCSC 表达 cTnT,与受体心肌相续。MCSC 移植组的梗死区周围微血管密度[(101.8±6.2)条/mm^2]大于对照组[(68.4±4.9)条/mm^2],与 MMSC 组[(97.2±3.2)条/mm^2]比较,差异无统计学意义。结论移植入心肌梗死模型的 MCSC 能够分化为功能性心肌,明显改善心功能,并诱导血管新生。MCSC 的移植治疗效果优于 MMSC。Objective To evaluate the therapeutic potential of marrow-derived cardiac stem cell （MCSC） transplantation after myocardial infarction （MI） in rats. Methods MCSC were selected from the marrow mesenchymal stem cell （MMSC） of male SD rats by single-cell cloning culture. MI was induced by left anterior descending artery ligating in female SD rats. Equal volume PBS, MMSC and MCSC were transplanted at the border zone of the infarct one week after MI. Cardiac function was assessed by echocardiography at four weeks after cell transplantation. The hearts were removed and morphological changes of scar tissue were examined with HE staining and Masson trichrome staining, VEGFR-1^＋ capillary vessels were labeled with immunohistochemical staining. Scar area and vessel density were measured by image analyzer. MCSC containing Y chromosome were examined using in situ fluorescent hybridization, and cardiomyocyte cTnT expression was also analyzed. Results Cardiac transcription factor Nkx2. 5 was expressed at low level in c-kit^＋ MCSC. Four weeks after cell transplantation, left ventricular fractional shortening and ejection fraction were significantly higher while scar area was significantly lower in MCSC group compared to MMSC group and control group, cTnT was expressed in cells containing Y chromosome and these cells were connected with myocardium of recipient rats in the rats transplanted with MCSC. Vessel density around the infarcted tissue in MCSC group was similar as that in MMSC group and significantly higher than that in control group. Conclusion MSCS could effectually differentiate into functional cardiomyocytes at the border zone of the infarct, and MCSC transplantation post MI significantly improved cardiac functions and promoted angiogenesis.

关 键 词：心肌梗塞 干细胞 骨髓 移植 

分 类 号：R542.22[医药卫生—心血管疾病]