骨髓间充质干细胞重复移植对大鼠脑梗死后bcl-2、caspase-3蛋白表达的影响

EFFECT OF REPEAT ADMINISTERED BONE MARROW MESENCHYMAL STEM CELLS ON EXPRESSION OF BCL-2 AND CASPASE-3 PROTEIN ON THE ISCHEMIC CEREBRAL INFARCTION IN RAT

目的探讨重复移植骨髓间充质干细胞(BMSCs)治疗大鼠脑梗死bcl-2、caspase-3蛋白表达。方法 BMSCs体外培养、纯化。线栓法建立大鼠永久性大脑中动脉阻塞模型,随机分为对照组(n=15):1,7d分别注射1ml生理盐水;单次组(n=15):1d,注射1ml细胞悬液,7d注射1ml生理盐水;重复组(n=15)在1,7d注射1ml细胞悬液,均尾静脉注射。神经功能评分进行行为学评价;TTC染色评估梗死体积;免疫组化及western检测bcl-2、caspase-3蛋白表达。结果单次组、重复组mNSS评分比对照组显著降低(P<0.01),同组比较评分降低(P<0.05);单次组、重复组相对梗死体积比对照组小(P<0.05),单次组和重复组bcl-2阳性细胞灰度值比对照组大(P<0.01),caspase-3阳性细胞灰度值降低(P<0.05),单次组和重复组bcl-2蛋白含量比对照组多(P<0.01),caspase-3蛋白含量降低(P<0.05)。结论重复移植BMSCs治疗脑梗死可更好改善大鼠神经功能,减少梗死体积,可能与上调bcl-2蛋白表达,减少caspase-3蛋白表达有关系。

Objective To investigate the expression of bcl-2 and caspase-3 protein on the ischemic cere-bral infarction in rat with repeat administered bone marrow mesenchymal stem cells（BMSCs ）.Methods In vitro cultivated and purified the BMSCs of the Sprague Dawley rats and established cerebral ischemia model with modified Zea Longa method.Forty-five rats were divided into three groups （control group, single group and repeat group）,15 rats in each group,and then treated respectively with intravenous injec-tion of saline 1ml 1d and 7d after MCAO or BMSCs 1ml 1d and saline 1ml 7d or BMSCs 1ml 1d and 7d. Cerebral infarction volume of rats was observed by TTC staining.The expression of bcl-2 and caspase-3 protein was detected through Immunohistochemistry and Western blot.Results Compared with the control group,in the single group and repeat group,the mNSS score was remarkable lower,the cerebral infaration volume was smaller,the expression of caspase-3 protein was also lower,but the expression of bcl-2 was higher（allP 〈0.05）.In addition,the above indicators were better in the repeat group than those in the sin-gle group（allP 〈0.05）.Conclusion Repeat transplantation of BMSCs after stroke in rats can improve the neurological function,decrease the infarction volume and the expression of caspase-3 and increase the ex-pression of bcl-2.