骨髓间充质干细胞移植对脑梗死小鼠大脑HMGB1释放的影响

发布时间：2018-06-24 03:32

本文选题：骨髓间充质干细胞 + 脑梗死　；参考：《郑州大学》2016年硕士论文

【摘要】：背景及目的脑梗死已成为中国人的第二位死亡原因,仅次于恶性肿瘤。随着我国逐步进入老龄化社会,脑梗死已经成为一个重大的社会问题。脑梗死早期炎症反应的发生是导致神经元死亡、机体神经功能缺失的重要原因,抑制脑梗死早期脑组织炎症的发生和发展能够有效改善脑梗死后神经功能缺损症状。骨髓间充质干细胞(Bone Marrow Stromal Cells,BMSCs)是脑梗死最有前景的干细胞之一,其可通过抑制炎症反应发挥其保护作用,但其具体机制尚未被完全阐明。HMGB1是脑梗死早期炎症发生的重要起始因子,因此考虑抑制HMGB1的释放是其炎症抑制机制之一,故设计本实验予以验证。方法Ficoll淋巴细胞分离液梯度离心法收集小鼠骨髓单个核细胞,体外培养分离传代得到BMSCs,鉴定后低温保存;之后通过线拴法制作小鼠脑梗死模型(t MCAO),按照研究要求将小鼠随机分为3组:假手术组,溶剂组和BMSCs移植组;尾静脉注射移植BMSCs至移植组小鼠体内,使用TTC染色法,观察建模情况及BMSCs对小鼠脑梗死体积的影响;各组小鼠分别进行m NSS评分,评价其神经功能缺损症状;然后通过免疫荧光技术,观察小鼠大脑梗死周围区HMGB1的释放情况;Western Blot技术检测脑组织TNF-α、IL-1β等炎症因子、HMGB1及Bcl-2等蛋白的表达情况,进而综合分析BMSCs对HMGB1释放的影响,阐述其对脑梗死小鼠早期炎症反应的作用机制。结果静脉移植BMSCs能够有效减少小鼠脑梗死脑组织体积及梗死后脑组织TNF-α、IL-1β等炎症因子的表达;显著改善梗死小鼠神经功能缺损症状并增加抗凋亡因子Bcl-2的表达;与溶剂组相比,移植组小鼠大脑脑组织HMGB1释放较溶剂组更少。结论BMSCs移植能通过抑制梗死早期小鼠脑组织炎症反应的进展,显著改善脑梗死小鼠的神经功能缺损症状,这一治疗作用可能与其抑制脑组织中HMGB1的释放有关。
[Abstract]:Background and objective Cerebral infarction has become the second leading cause of death in Chinese, second only to malignant tumors. As our country gradually enters the aging society, cerebral infarction has become a major social problem. The occurrence of inflammatory reaction in the early stage of cerebral infarction is an important cause of neuronal death and loss of nervous function. Inhibiting the occurrence and development of inflammation in brain tissue in the early stage of cerebral infarction can effectively improve the symptoms of neurological deficit after cerebral infarction. Bone Marrow Stromal cells (BMSCs) is one of the most promising stem cells in cerebral infarction. It can play a protective role by inhibiting inflammatory response, but its specific mechanism has not been fully elucidated that .HMGB1 is an important initial factor of inflammation in the early stage of cerebral infarction. Therefore, one of its inflammatory inhibition mechanisms is to consider inhibiting the release of HMGB1, so this experiment is designed to verify it. Methods Mouse bone marrow mononuclear cells were collected by gradient centrifugation with Ficoll lymphocyte isolate. BMSCs were isolated and subcultured in vitro. The mice were randomly divided into three groups: sham operation group, solvent group and BMSCs transplantation group. To observe the model and the effect of BMSCs on cerebral infarction volume in mice; to evaluate the neurological deficit symptoms of mice in each group by mNSS scores; and then to evaluate the neurological deficit symptoms by immunofluorescence technique. To observe the release of HMGB1 in the cerebral infarction area of mice and to detect the expression of HMGB1, Bcl-2 and other inflammatory factors such as TNF- 伪 and IL-1 尾 in brain tissue by Western blot, and then to analyze the effect of BMSCs on HMGB1 release. The mechanism of its effect on early inflammatory reaction in mice with cerebral infarction was described. Results Venous transplantation of BMSCs could effectively reduce the volume of cerebral tissue and the expression of inflammatory factors such as TNF- 伪 and IL-1 尾 in the cerebral tissue of mice with cerebral infarction, and significantly improve the symptoms of neurological deficit and increase the expression of anti-apoptotic factor Bcl-2 in the infarcted mice. HMGB1 release from brain tissue of transplanted mice was less than that of solvent group. Conclusion BMSCs transplantation can significantly improve the neurological deficit symptoms of cerebral infarction mice by inhibiting the progress of inflammatory reaction in cerebral tissue in early stage of infarction. This therapeutic effect may be related to the inhibition of HMGB1 release in brain tissue.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R743.3

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