骨髓间充质干细胞抑制大鼠脑缺血后神经细胞凋亡的机制研究

发布时间：2018-03-07 14:09

本文选题：骨髓间充质干细胞　切入点：脑缺血/再灌注　出处：《吉林大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:本实验主要探讨骨髓间充质干细胞对发生脑缺血/再灌注损伤的大鼠,在脑梗死体积、神经功能损伤评分、脑缺血区域细胞的凋亡情况等方面的影响,并对其通过抑制神经细胞凋亡,从而对神经细胞发挥保护作用的可能机制给予研究。方法:对大鼠骨髓进行初步分离,细胞培养,骨髓间充质干细胞鉴定,最终标记等步骤,从而获取实验用骨髓间充质干细胞;将SD雄性大鼠60只进行分组,随机平均分为模型组、实验组、假手术组,共设3组,20只每组。假手术组不行栓塞大脑中动脉处理,其余组利用线栓栓塞大鼠大脑中动脉,缺血90分钟后,取出线栓,造成大鼠脑缺血再灌注,从而建立大鼠脑缺血再灌注模型,再灌注时间为24 h。实验组给予浓度为1×109/L的骨髓间充质干细胞1ml,采用尾静脉注射法,另两组经尾静脉给予1ml生理盐水。分别在术后第1 Day、第1Week、第2 Weeks进行神经功能评分;在术后第2周测定脑梗死体积;RT-PCR法及Western blot检测Bax/Bcl-2的m RNA和蛋白比例;检测caspase-3蛋白含量、检测Survivin蛋白表达情况。结果:与模型组比较,实验组神经功能评分显著降低(P0.05),脑梗死体积显著减小(P0.05),Bcl-2的m RNA和蛋白表达明显升高;而Bax的m RNA和蛋白表达明显降低(P0.05),caspase-3蛋白表达减少(P0.05),Survivin蛋白表达增加(P0.05)。结论:1、骨髓间充质干细胞具有保护神经细胞的作用,可以减少脑梗死体积。2、骨髓间充质干细胞能够减轻缺血灌注后神经功能的损伤,促进神经功能的恢复。3、上调Bcl-2m RNA的表达,促进Bcl-2蛋白表达。4.降低Baxm RNA的表达,减少Bax蛋白含量。5.骨髓间充质干细胞能够增加Survivin蛋白的表达,减少caspase-3蛋白的表达。综上所述,骨髓间充质干细胞能抑制神经细胞凋亡,提高神经细胞对缺血再灌注损伤的耐受力,发挥神经保护作用。总之骨髓间充质干细胞能够有效抑制缺血再灌注后神经细胞的凋亡,但其更为详细的机制仍需进一步的深入研究。
[Abstract]:Objective: to investigate the effects of bone marrow mesenchymal stem cells (BMSCs) on cerebral infarction volume, neurological function injury score, apoptosis in ischemic area of brain in rats with cerebral ischemia / reperfusion injury. The possible mechanism of its protective effect on nerve cells was studied by inhibiting neuronal apoptosis. Methods: the primary isolation of rat bone marrow, cell culture, identification of bone marrow mesenchymal stem cells, and final labeling, etc. In order to obtain bone marrow mesenchymal stem cells for experiment, 60 male SD rats were divided into three groups: model group, experimental group and sham operation group. In the other group, the middle cerebral artery was embolized with thread embolism. After 90 minutes of ischemia, the thread embolus was removed to make the cerebral ischemia-reperfusion in rats, and the model of cerebral ischemia-reperfusion was established. The time of reperfusion was 24 hours. The experimental group was given 1 ml of bone marrow mesenchymal stem cells (1 脳 10 9 / L) by caudal vein injection, and the other two groups were given 1 ml of normal saline through tail vein. The neurological function was evaluated at 1 day, 1 week and 2 Weeks after operation. At the second week after operation, the ratio of m RNA and protein of Bax/Bcl-2 was detected by RT-PCR and Western blot, and the content of caspase-3 protein and the expression of Survivin protein were detected. Results: compared with the model group, the ratio of m RNA and protein of Bax/Bcl-2 was detected by RT-PCR and Western blot at the second week after operation. In the experimental group, the scores of nerve function decreased significantly (P 0.05), and the volume of cerebral infarction significantly decreased the expression of m RNA and protein of Bcl 2. The expression of m RNA and protein in Bax decreased significantly, while the expression of caspase-3 decreased significantly. Conclusion: bone marrow mesenchymal stem cells (BMSCs) can protect nerve cells by increasing the expression of survivin. Bone marrow mesenchymal stem cells can reduce the injury of nerve function, promote the recovery of nerve function, up-regulate the expression of Bcl-2m RNA, promote the expression of Bcl-2 protein, and decrease the expression of Baxm RNA. Bone marrow mesenchymal stem cells can increase the expression of Survivin protein and decrease the expression of caspase-3 protein. In conclusion, bone marrow mesenchymal stem cells can inhibit neuronal apoptosis. In short, bone marrow mesenchymal stem cells can effectively inhibit neuronal apoptosis after ischemia-reperfusion injury. However, its more detailed mechanism still needs to be further studied.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R743.3

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