经静脉移植骨髓基质细胞治疗大鼠局灶性脑梗死Bcl-2蛋白表达的实验研究

Intravenous administration of bone marrow stromal cells after stroke in rats and the expression of Bcl-2 protein

目的 观察经静脉移植骨髓基质细胞(MSCs)治疗大鼠局灶性脑梗死时bcl- 2蛋白的表达。方法 体外培养及扩增骨髓基质细胞后,将实验大鼠2 4只分为3组:第1组(n=8)单纯大脑中动脉闭塞(MCAO)组;第2组(n=8) ,MCAO后4 8h经尾静脉注射生理盐水1ml;第3组(n=8) ,MCAO后4 8h经尾静脉注射1×10 7个MSCs。神经功能严重程度评分日一次进行。所有动物均在MCAO后第8天处死。用免疫细胞化学方法体外鉴定骨髓基质细胞;免疫组织化学染色显示鼠脑内bcl- 2蛋白表达情况。结果 原代培养2 0 d后可分离得到骨髓基质细胞,在5代以前生长形态相对稳定。与对照组相比,实验组大鼠的神经功能严重程度评分有明显改善(P<0 .0 0 1) ,且脑内bcl-2表达增加(P<0 .0 0 1)。结论 经静脉移植骨髓基质细胞可明显改善大鼠局灶性脑梗死后神经功能恢复,并可上调脑内bcl- 2蛋白表达水平。

Objective To observe the expression of Bcl-2 protein during the course of intravenous administration of bone marrow stromal cells after stroke in rats.Methods In vitro cultivate and amplificate the bone marrow stromal cells of the Wistar rats.Separate 24 rats into three groups:Group 1(n=8) middle cerebral artery occlusion (MCAO);Group 2(n=8)intravenous injection of saline 1ml 48 hours after MCAO;Group 3(n=8) intravenous transplantation of bone marrow stromal cells 1×10 7.Observe the nervous severity score of every rat everyday.All animals were put to death 8 days after MCAO.In vitro identify the MSCs by immunocytochemical method and display the expression of bcl-2 protein by immunohistochemistry stain.Results Bone marrow stromal cells can be separated 20 days after primary culture.The living characteristic of the cultured cells was quite stable before the fifth generation.In contrast to the control group,the NSS of the test group ameliorated significantly(P<0.001) and the expression of bcl-2 protein improved greatly (P<0.001).Conclusion Intravenous administration of bone marrow stromal cells after stroke in rats can promote the recovery of the nervous functions and improve the expression of bcl-2 protein.