兰州大学机构库 >生命科学学院
CSF1R的抑制剂Ki20227对全脑缺血小鼠小胶质细胞和神经元的影响
Alternative TitleKi20227 influences the morphology of microglia and neurons through inhibition of CSF1R in mice during global ischemia
侯博儒
Thesis Advisor张胜祥
2016-12-16
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name博士
Keyword中风 小胶质细胞 重灌流 Ki20227 树突棘
Abstract

脑中风严重危害人类的生命健康。缺血性脑中风指由各种原因所致的大脑血流供应障碍,从而导致神经元、胶质细胞以及周围血管因局灶性脑缺血而受损的急性脑血管事件的一种类型,相应的神经功能缺损包括偏瘫、失语、认知和学习能力受损等。目前中风治疗方法主要是通过多种机制来提高神经环路的可塑性或者修复神经环路,新的方法需要关注在组织修复过程中多种细胞之间相互联系的复杂性和细胞的存活途径。中风后,小胶质细胞,血管内皮细胞以及一些其他诱导因素共同协调参与组织损伤和修复。小胶质细胞是脑内具有免疫活性的细胞,在不同的生理病理条件下具有多种多样的功能。脑中风后,小胶质细胞迅速反应,目前对脑中风后小胶质细胞的功能还有很大争议。小胶质细胞对神经元具有两种作用:积极作用和消极作用,但在中风后,小胶质细胞的作用还不是很清楚,同时小胶质细胞增生的调控机制也尚不清楚。本研究探讨中风后Ki20227对小胶质细胞和神经元的影响,以及中风后小胶质细胞是否对神经元具有保护作用。

本论文利用小胶质细胞特异性表达绿色荧光蛋白的转基因小鼠,采用药物Ki20227抑制CSF1R来干预小胶质细胞,通过建立全脑缺血脑中风模型,利用双光子活体成像技术,免疫组织化学染色,荧光显微成像分析,荧光定量PCR和行为学测试等研究方法,揭示中风后小胶质细胞对神经元的影响,初步探索小胶质细胞增生的调控机制。我们的结果表明,中风后BrdU染色显示小胶质细胞存在增殖现象,中风导致小胶质细胞中细胞因子表达异常;神经元数目减少;皮层和海马树突棘密度下降;行为学异常。同时发现全脑缺血后采用药物Ki20227抑制CSF1R后,一方面导致了小胶质细胞密度下降、突起减少、相关细胞因子表达异常,同时增殖的小胶质细胞密度下降,而另一方面进一步导致了树突棘密度的下降,突触分子表达水平下降,以及行为学异常。综上,我们的结果揭示了小胶质细胞的形态和细胞群密度以及相关细胞因子的表达与CSF1R密切相关;小胶质细胞在全脑缺血后对神经元具有保护作用。

Other Abstract

The stroke causes serious damage to people’s health. Ischemic stroke is a type of acute cerebrovascular incidence in which blood supply obstacles in the brain caused by various reasons damage neurons, microglia and surrounding vessels due to focal cerebral ischemia. The corresponding neurologic impairments include hemiplegia, aphasias, the cognitive and language impairments. At present, the main treatment for stroke is focused on improving the plasticity of neural circuits or on their repairing through multiple mechanisms. The new method should focus on complexity of the connections among multiple cells as well as cell survival pathways in the tissue repair process. After the stroke, microglia together with endothelial cells and other inducing factors, take part in the renovation of damaged tissues. As immunological competent cells in the brain, microglia can respond rapidly after the stroke. Under different physiologic and pathologic conditions, they have different functions. But there is still much debate as to their function after the stroke currently. Microglia may exert positive or negative effect on the neurons. At present, however, their effects after the stroke, along with the regulatory mechanism of microglial proliferation, are still unclear. In present study, we investigated the effect of Ki20227 on microglia and neurons, and whether microglia can improve the survival of neurons after global ischemia.

This research is based on the experiment on the transgenic mice whose microglia can specifically express the green fluorescent proteins. Microglia were intervened with by inhibiting colony-stimulating factor 1 receptor (CSF1R) with Ki20227 and a global ischemic stroke model was established. Using two-photon transcranial imaging technique, the immunohistochemical staining technique, fluorescence microscopy, fluorescence quantitative polymerase chain reaction, and the behavioral testing, this research tries to reveal the influence of microglia on neurons after the stroke and to tentatively explore the regulatory mechanism of microglial proliferation. BrdU (5-Bromo-2-deoxy Uridine) staining demonstrated that microglia proliferated after stroke, and the results showed that stroke caused the abnormal expression of cytokines in the microglia, the reduction in the number of neurons, and the decline in the density of dendritic spines in cortex and hippocampus. In addition, stroke also resulted in behavior abnormality. In the meantime, the inhibition of CSF1R by Ki20227 led to a decrease in the number of microglia and cell processes, and reduction in proliferative microglia. The relevant cytokines were also abnormally expressed. In addition, the density of neurons and dendritic spines declined, the expression level of synaptic molecules decreased and abnormal behaviors occured. To sum up, the results revealed that the morphology of the microglia, the density of microglia populations, and the expression of relevant cytokines were closely related to CSF1R. Microglia may have a protective role for neurons after global ischemia.

URL查看原文
Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/222020
Collection生命科学学院
Recommended Citation
GB/T 7714
侯博儒. CSF1R的抑制剂Ki20227对全脑缺血小鼠小胶质细胞和神经元的影响[D]. 兰州. 兰州大学,2016.
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