摘要
目的:外周血干细胞趋向成熟细胞,分化为其他细胞的潜能较小,需在动员剂参与下移植发挥其干细胞分化作用。观察将自体外周血单个核细胞作为移植细胞,在动员剂参与下的成心肌样细胞形成和促进微血管生成的作用。方法:实验于2003-04/2004-03在哈尔滨医科大学附属二院动物实验中心和病理科完成。选用60只日本大耳白兔,随机分成3组,移植组和动员剂组及对照组,每组20只。移植组和动员剂组结扎冠状动脉前降支建立心肌梗死模型,而后连续给予粒细胞集落刺激因子7d,移植组在模型建立后1周,将5-溴-2'-脱氧脲苷标记的细胞悬液心外膜注射到心肌梗死及邻近区,动员剂和对照组只给予等量的生理盐水注射。以苏木精-伊红染色发现5-溴-2'-脱氧脲苷阳性细胞和肌动蛋白表达阳性(为移植后成活为心肌样细胞的标志)。以Masson和免疫组化染色检测移植的自体外周血单个核细胞存活及分化情况,评估移植后细胞成活的组织学变化,并测定含梗死区心肌的毛细血管密度。结果:按意向处理分析,进入结果分析的白兔60只,每组20只。①移植后4周,移植组免疫组化染色在梗死区可发现5-溴-2'-脱氧脲苷阳性细胞,且表达肌动蛋白阳性。②苏木精-伊红染色显示对照组梗死区结构紊乱,而动员剂和细胞移植组梗死区细胞排列有序,同时动员剂和细胞移植组可见大量的新生毛细血管,其密度明显高于对照组犤(58.2±11.5),(52.3±6.0),(21.6±4.9)个/高倍视野,P<0.01)犦。结论:在动员剂参与下,自体外周血单个核细胞移植后4周可以在梗死区检测到5-溴-2'-脱氧脲苷阳性细胞,且心肌特异性蛋白肌动蛋白表达阳性,在一定程度上证实了植入的细胞可存活并分化成心肌样细胞,同时促进了心肌局部的毛细血管生成。
AIM: Peripheral blood stem cells tend to mature cells,and the potential of differentiating into other cells is nearly inpossible.Therefore,the stem cells play the differentiating role by means of transplantation concerned with mobilizing agent. This paper observes the formation of cardioblasts and the role of promoting the microvascularization involved by mobilizing agent with autologous peripheral blood mononuclear cells as the transplanted cells. METHODS: The experiment was carried out in the animal experimental center and Department of Pathology, the Second Affiliated Hospital of Harbin Medical University from April 2003 to March 2004. Sixty Japanese big-ear rabbits were randomly divided into 3 groups with 20 rabbits in each group: transplantation group, mobilizing agent group and control group. Myocardial infarction models were made by ligating the anterior descending coronary artery in the transplantation group and mobilizing agent group, and then they were treated with granulocyte colony-stimulating factors for continuous 7 days; One week after model establishment in the transplantation group, cell suspensions labeled with BrdU were injected into the infarcted regions and vicinities; The rabbits in the mobilizing agent group and control group were injected with saline of the same volume. BrdU positive cells and positive expression of actin, which were taken as the markers of survival of myocardium-like cells after transplantation, were observed with haematine-eosin staining. The survival and differentiation of transplanted autologous peripheral blood mononuclear cells were detected with Masson and immunohistochemical staining; The histological changes of survived cells after transplantation were assessed, and the capillary densities of myocardium containing infarcted area were determined. RESULTS: According to intention-to-treat analysis, 60 white rabbits were involved in the analysis of results with 20 in each group. ①Four weeks after transplantation, BrdU positive cells and positive expression of actin were observed in the infarcted area with immunohistochemical staining in the transplantation group.②Haematine-eosin staining showed that the structure of the infarcted area was disorder in the control group, and the cells in the infarcted area were orderly arranged in the mobilizing agent group and transplantation group; Meanwhile, plenty of newly generated blood capillaries were observed in the mobilizing agent group and transplantation group, the densities were obviously higher than that in the control group[(58.2±11.5), (52.3±6.0), (21.6±4.9) per high power visual sight, P< 0.01]. CONCLUSION: Through the participation of mobilizing agent,BrdU positive cells can be detected in the infarcted area and the myocardial specific protein actin expresses positively 4 weeks after the autologous peripheral blood mononuclear cells transplantation; It is proved to a certain degree that the transplanted cells can survive and differentiate into myocardium-like cells, and promotes the generation of blood capillary in local myocardium.
出处
《中国临床康复》
CAS
CSCD
北大核心
2005年第22期60-61,i002,共3页
Chinese Journal of Clinical Rehabilitation
