大鼠骨髓间质干细胞和新鲜骨髓细胞静脉移植治疗脑梗死的疗效.doc

1 大鼠骨髓间质干细胞和新鲜骨髓细胞静脉移植治疗脑梗死的疗效【摘要】目的比较大鼠骨髓间质干细胞(rMSC) 和新鲜骨髓细胞(fBMC) 静脉移植后在脑梗死时脑内存活、迁移及分化情况, 及对神经功能恢复的影响。方法制备大鼠脑梗死模型, 经尾静脉将标记的 rMSC 和 fBMC 植入体内, 于不同时程对神经功能状况进行评分。应用荧光激发及免疫组织化学方法检测移植细胞在脑内存活、迁移及分化情况。细胞化学染色观察 rMSC 体外分化情况。结果与对照组比较, 两移植组神经功能改善明显(), 组间 mNSS 评分比较无统计学意义() 。两移植组在荧光激发下均可见移植细胞在梗死区及其周边区聚集并存活, 且有部分植入细胞分化表达神经细胞表面标志物, 在体外 rMSC 未见类似分化表达。两组植入细胞存活与分化标志表达率差别无统计学意义。结论 rMSC 和 fBMC 经静脉移植后在宿主脑内均能够存活、迁移并分化表达神经细胞表型, 均有促进脑梗死后神经功能恢复的作用。【关键词】间质干细胞移植; 骨髓细胞; 脑梗死; 疾病模型, 动物 2 ABSTRACT: Objective To investigate that rat mesenchymal stem cells(rMSC) and fresh bone marrow cells(fBMC) infused intravenously for migrate to infarct focus, survive, differentiate into nerve cells, and improve neurological function after cerebral ischemia in adult rats. Methods rMSC were cultured in vitro. Adult male SD rats were subjected to transient(2 hours) middle cerebral artery occlusion(MCAO). rMSC or fBMC or PBS were infused via tail vEin at 24 hour after MCAO respectively. Functional measurements using the modified neurological severity scores(mNSS) were performed at 24 hour, 1 week and 2 weeks after MCAO respectively. Immunohistochemical staining and laser scanning microscopy were used to identify rMSC and cells derived from bone marrow in the brain sections. The specific marking protEIn of neurons and glial cell were dectected by immunocytochemistry in vitro. Results rMSC proliferated rapidly in vitro. There was significant neurological function improvement in rats treated with rMSC and pared with that of control group, as evidenced by mNSS scores(). Theres not significant difference in mNSS scores between rMSC and fBMC treated 3 groups. rMSC and fBMC survived and were localized around the ischemic core and its boundary zone, and a few cells expressed Nestin, NF-200 and GFAP, but there were no expression when rMSC cultured in vitro. Conclusion Both rMSC and fBMC can survive and localize in the ischemic area of the brain, and some cells expressed the phenotypic protein marker of ne