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目的探讨骨髓间充质干细胞(MSCs)移植治疗脑缺血模型大鼠的时间窗及最佳移植时间点。方法雄性SD大鼠56只,随机分为移植组32只和对照组24只,在建立大鼠大脑中动脉缺血2 h再灌注模型后分为1、7、14、21 d 4个时间点。对大鼠进行改良神经功能缺损评分、HE染色、Bielshowsky Iuxol Fast Blue染色、直接免疫荧光染色和免疫组织化学染色。结果移植组1、7 d在移植14 d后,14、21 d在移植21 d后改良神经功能缺损评分明显低于对照组(P<0.05)。移植组1、7 d胶质纤维酸性蛋白阳性细胞数、突触素累计吸光度值、胼胝体面积明显高于对照组(P<0.05)。移植组1、7、14、21 d Ki-67阳性细胞数明显高于对照组(P<0.05)。移植组7 d神经元特异性烯醇酶累计吸光度值明显高于对照组(P<0.05)。移植组1、7 d Nogo A阳性细胞数明显低于对照组(P<0.05)。结论 MSCs移植治疗大鼠脑缺血模型的时间窗可延长至制模后的21 d,最佳移植时间点为制模后7 d。MSCs移植治疗脑梗死的作用机制可能与其促进内源性修复相关。
Objective To investigate the time window and optimal transplantation time of bone marrow mesenchymal stem cells (MSCs) transplantation in rats with cerebral ischemia. Methods Fifty-six male Sprague-Dawley rats were randomly divided into two groups: transplantation group (32 rats) and control group (24 rats). After establishment of rat middle cerebral artery ischemia 2 h reperfusion model, rats were divided into 4 groups . Rats were evaluated for neurological deficit score, HE staining, Bielshowsky Iuxol Fast Blue staining, direct immunofluorescence staining and immunohistochemical staining. Results On the first day after transplantation, the improved neurological deficit scores were significantly lower in the transplantation group than those in the control group (P <0.05) after 14 days of transplantation and 14 and 21 days after transplantation. The number of glial fibrillary acidic protein positive cells, synaptophysin cumulative absorbance value, and corpus callosum area in the transplantation group at 1,7 d were significantly higher than those in the control group (P <0.05). The number of Ki-67 positive cells on the 1st, 7th, 14th and 21st day in the transplantation group was significantly higher than that in the control group (P <0.05). The cumulative value of neuron-specific enolase in the 7-day transplantation group was significantly higher than that in the control group (P <0.05). The number of Nogo A positive cells in the transplantation group at 1 and 7 days was significantly lower than that in the control group (P <0.05). Conclusion The time window of MSCs transplantation in rat model of cerebral ischemia can be prolonged to 21 days after model establishment, and the best transplantation time point is 7 days after model establishment. The mechanism of MSCs transplantation in the treatment of cerebral infarction may be related to its promotion of endogenous repair.