论文部分内容阅读
目的探讨猪心肌缺血再灌注损伤区自体骨髓单个核细胞(BM-MNC)和外周血内皮祖细胞(EPC)移植后细胞存活和分化情况。方法 23头中国小型猪随机分为 BM-MNC 组(n=9)、EPC 组(n=7)和对照组(n=7)。BM-MNC 组,将 PKH26标记的 BM-MNC 经冠状动脉移植于心肌缺血再灌注损伤区。EPC 组,将 CM-DiI 标记的 EPC 经冠状动脉移植于心肌缺血再灌注损伤区。对照组,磷酸盐缓冲液对照。4周后观察荧光标记的移植细胞,并在缺血再灌注区内取材进行病理染色,用兔抗Ⅷ因子相关抗原多克隆抗体,鼠抗兔α-肌小节-肌动蛋白抗体和兔抗人纤连蛋白多克隆抗体免疫组化染色,比较各组缺血再灌注损伤区血管再生情况和反应强度。选取免疫组化可疑分化的组织观察可疑分化细胞的超微结构。结果缺血再灌注区的小血管数目 BM-MNC 组为(13.39±6.96)个/高倍视野,EPC 组为(12.39±4.72)个/高倍视野,两者问差异无统计学意义,而高于对照组的(3.50±1.90)个/高倍视野(P<0.05)。BM-MNC 和 EPC 组兔抗人纤连蛋白多克隆抗体阳性反应强度显著低于对照组(P<0.05),而鼠抗兔α-肌小节-肌动蛋白抗体阳性反应强度三组间差异无统计学意义。BM-MNC 组1头猪的密集细胞团形态结构上结合免疫组化染色鉴定,可能是干细胞的原位生长的表现。电镜检查发现了比较幼稚的内皮细胞和心肌样细胞。结论 EPC 和 BM-MNC 移植于梗死区具有刺激小血管形成、抑制纤维化的作用,还可能向心肌细胞方向分化。
Objective To investigate the cell survival and differentiation of autologous bone marrow mononuclear cells (BM-MNC) and peripheral blood endothelial progenitor cells (EPC) after myocardial ischemia-reperfusion injury in pigs. Method 23 Chinese miniature pigs were randomly divided into BM-MNC group (n = 9), EPC group (n = 7) and control group (n = 7). BM-MNC group, PKH26-labeled BM-MNC was transplanted into myocardial ischemic reperfusion injury area via coronary artery. EPC group, EPCs labeled with CM-DiI were transplanted into myocardial ischemic reperfusion injury area via coronary artery. Control group, phosphate buffer control. Four weeks later, the fluorescent labeled cells were observed and harvested for histopathological examination in ischemic reperfusion area. Rabbit anti-Ⅷ factor-related antigen polyclonal antibody, mouse anti-rabbit α-sarcomeric actin antibody and rabbit anti-human The fibronectin polyclonal antibody was immunohistochemically stained to compare the angiogenesis and reactivity of ischemic reperfusion injury in each group. Susceptible differentiated tissues were selected to observe the ultrastructure of suspicious differentiated cells. Results The number of small blood vessels in ischemia-reperfusion area was (13.39 ± 6.96) / high power in BM-MNC group and (12.39 ± 4.72) / high power in EPC group, but the difference was not significant The control group (3.50 ± 1.90) / high power field (P <0.05). The positive reaction intensity of anti-human Fibronectin polyclonal antibody in BM-MNC and EPC rabbits was significantly lower than that in control group (P <0.05), while the positive reaction intensity of anti-rabbit anti-rabbit anti-actin antibody Statistical significance. BM-MNC group 1 pigs dense cell mass morphology combined with immunohistochemical staining may be the performance of stem cells in situ growth. Electron microscopy revealed naive endothelial cells and cardiomyocyte-like cells. Conclusion EPC and BM-MNC transplanted in the infarct area can stimulate the formation of small blood vessels and inhibit fibrosis, and may also differentiate into cardiomyocytes.