目的探讨立体定向移植人骨髓间充质干细胞(hMSCs)治疗对局灶性脑缺血大鼠海马区α-微管蛋白表达的影响。方法体外培养、扩增hMSCs;100只大鼠随机分为正常对照(NC)组、假手术组、缺血再灌注(IR)组、假移植组、移植组,每组再分为1d、3d、7d、28d4个时间点;制作大鼠局灶性脑缺血90min再灌注1h的IR模型,制模成功后移植组大鼠立即进行立体定向hMSCs移植,移植后相应时间点应用免疫组化及免疫荧光染色,观察各组大鼠海马区α-微管蛋白表达。结果 NC组、假手术组大鼠各时间点海马区α-微管蛋白表达的差异无统计学意义;与之相比,IR组、假移植组各时间点及移植组移植后1d、3d时表达明显下降(均P〈0.01);但移植组各时间点海马区α-微管蛋白表达明显高于IR组及假移植组(均P〈0.01),移植7~28d时与NC组比较,差异无统计学意义。结论局灶性脑缺血大鼠海马区α-微管蛋白表达减少;hMSCs移植促进α-微管蛋白表达增加,这可能是其减轻脑缺血后神经损伤的机制之一。
Objective To explore the influence of human bone marrow mesenchymal stem cells (hMSCs) oriental transplant on expression of α-tubulin in hippocampus in rats with cerebral focal ischemia. Methods The hMSCs were cultured,purified,and proliferated in vitro. 100 Wistar rats were randomly divided into normal control (NC) group,sham-operation group,cerebral ischemia/reperfusion (IR) group,sham-transplant group and hMSCs transplant group; each group was including 4 time points (after transplant 1 d,3 d,7 d and 28 d). The IR models were prepared by middle cerebral artery occlusion 90 min and reperfusion 1 h. Then the hMSCs were orientatal transplanted immediately. Immunohistochemical and immunofluorescence methods were used to detect the expressions of α-tubulin in hippocampus. Results The expression of α-tubulin in hippocampus between the NC group and the shame-operation group in each time point was no significant difference. Compared with them,the expressions of α-tubulin in IR group,sham-transplant group on each time point and transplant group 1 d,3 d were significantly decreased (all P0.01). Compared with IR group and sham-operation group,the expression of α-tubulin in the transplant group in each time point were obviously increased (all P0.01). After transplanted 7-28 d,the difference of expression of α-tubulin in hippocampus was no statistical significance between transplant group and NC group. Conclusions The expression of α-tubulin in hippocampus is decreased in rats with cerebral focal ischemia. Transplanted hMSCs can improved the expression of α-tubulin. Which may be one of mechanism of that hMSCs can alleviate the neural injury after cerebral ischemia.