猪心肌缺血再灌注损伤区自体骨髓单个核细胞和外周血内皮祖细胞移植后细胞分化探讨  

Autologous bone marrow mononuclear cells and peripheral endothefial progenitor cells differentiation in myocardial ischemia reperfusion injury region in Swine

作　　者：李崇剑[1] 高润霖[1] 杨跃进[1] 宋来凤[2] 阮英茆[2] 胡奉环[1] 杨伟宪[1] 陈纪林[1] 乔树宾[1] 秦学文[1] 刘玉清[3] 陈在嘉[1] 

机构地区：[1]中国医学科学院中国协和医科大学心血管病研究所阜外心血管病医院冠心病诊治中心,北京100037 [2]中国医学科学院中国协和医科大学心血管病研究所阜外心血管病医院病理科,北京100037 [3]中国医学科学院中国协和医科大学心血管病研究所阜外心血管病医院中德分子医学实验室,北京100037

出　　处：《中华心血管病杂志》2007年第4期350-353,共4页Chinese Journal of Cardiology

摘　　要：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 differentiation status of autologous bone marrow mononuclear cells （ BM-MNC ） and peripheral endothelial progenitor cells （ EPC ） transplanted into myocardial ischemia reperfusion injury region in swine. Methods BM-MNC marked with PKH26 （ n = 9 ）, EPC marked with CM-DiI （n = 7 ）, phosphate buffer saline （control, n = 7 ） were transplanted into myocardial ischemia reperfusion injury region of swine by intracoronary artery injection. Specimens were harvested 4 weeks after injection for histological analysis （HE, immunochemical stain for vWF, α- sarcomeric-actin and fibronectin antibody ）. Cell differentiation was observed under transmission electromicroscope. Results The number of small blood vessels was similar between BM-MNC group and EPC group （13.39 ± 6. 96/HP vs. 12.39 ± 4. 72/HP, P 〈 0. 05 ）, but was significandy higher than that of control group （ P 〈 0. 05 ） . Responsive intensity of immunochemical stain for fibronectin antibody was significantly lower in BM-MNC and EPC groups than that in control group. Responsive intensity of immunochemical stain for α-sarcomeric-actin antibody was similar among the three groups. Cluster cells were observed in one swine from BM-MNC group which might relate to the proliferation of stem cells in situ. Immature endothelial cells and myocytes were also detected by transmission electromicroscope in BM-MNC and EPC group, Conclusion BM-MNC and EPC transplanted into myocardial ischemia reperfusion injury region in swine stimulated the formation of blood vessels and inhibited fibrogenesis.

关 键 词：再灌注损伤 干细胞 细胞移植 细胞分化 

分 类 号：R541[医药卫生—心血管疾病]