以清除和再殖方法揭示小胶质细胞在青光眼病程中的功能变化

Glaucoma could cause retinal ganglion cell (RGC) irreversible degeneration and severely affects the visual acuity of the patients. During this process, microglial activation plays a key role: both apoptosis and protection. The M2 (alternative activation) type of retinal polarization might exert a protective role of RGCs, which might be applied in the clinical field in near future...In this project, we will begin with the profile of the phenomenon (profiling the general picture of microglial polarization and RGC degeneration at different disease stage after AOH); the focal point will be the exploration of the mechanism (with the help of microglial depletion and on schedule repopulation, we investigate the different function of microglia at different polarized state, and also the possible molecular mechanism and signaling pathway);the orientation will be the discovery of the therapeutic strategy (select the key molecules which can delay the degeneration of RGCs though the regulation of microglial polarization; providing the scientific basis and potential therapies for the clinical interventions which take microglia as the therapeutic target...In this project, we determine to partly solve the following scientific problem with the combination of morphological, molecular biological method, RNA-seq and vision functional analysis: (1) what is the relationship between microglial polarization and RGC degeneration? (2) during the neurodegenerative diseases, what is the dynamic interaction between microglial activation and the inflammatory environment? And though what kind of critical molecules? (3) Can we treat the intractable neurodegenerative disease, such as glaucoma, by the modulation of microglial polarization?

青光眼导致不可逆的视网膜神经节细胞损伤，严重危及病人视力；此过程中小胶质细胞发挥关键性调控作用，特别是其M2型极化可能对节细胞起保护作用，且有临床应用的可能。本项目以现象描述为起始（绘制AOH模型中，小胶质细胞极化和RGC凋亡的全景图）；以机制探索为重点（通过小胶质细胞的清除和择期回归，深入探讨不同极化方式的小胶质细胞的不同功能及相关的分子机制）；以治疗策略研发为导向（筛选通过调控小胶质细胞极化来延缓RGC凋亡的关键分子，为“以小胶质细胞作为治疗靶点”的神经保护策略，提供科学依据和备选药物）。主要依靠形态学、分子生物学、转录组测序，视觉功能分析等手段，解答以下科学问题：小胶质细胞的极化状态与RGC的凋亡速率之间是怎样的关系？小胶质细胞极化与炎性微环境，在神经退行性疾病中，是怎样的动态互动关系，由哪些关键分子参与？能否通过调控小胶质细胞的极化状态，治疗神经退行性疾病，解决青光眼治疗难题？

青光眼所致的不可逆的视网膜神经节细胞损伤严重危及病人视力。.小胶质细胞是中枢神经系统内重要的免疫细胞，与视网膜神经退行性病变密切相关。它的活化和极化对各类神经元的凋亡和保护均发挥重要作用。.前期研究中证实小胶质细胞的活化和极化对青光眼中RGC的凋亡有重大影响，小胶质细胞活化对视网膜神经细胞的凋亡起关键调控作用，尤其是前期结果表明，视网膜内小胶质细胞的替代激活（M2）对RGC同样可能起到保护作用，且这种作用有指导临床的可能。..方法: 制作急性高眼压（Acute ocular hypertension，AOH）青光眼模型，小胶质细胞发生活化，通过形态学观察，统计在AOH青光眼模型中，不同时间节点的RGC存活数量，绘制不同时间节点下RGC存活数量曲线图；对PLX+AOH模型组采取PLX5622随意喂养清除小胶质细胞，CD+AOH模型组用CD粮食随意喂养，根据存活数量曲线图，选择在PLX+AOH模型组和CD+AOH模型组中，小鼠模型制作后7天，通过形态学观察，对RGC存活数量进行统计及分析，并用电生理和视动学进行视功能学检测清除小胶质细胞后，视网膜功能的改善情况。..结果：在AOH模型中，小胶质细胞呈高度活化状态，促进RGC死亡，RGC存活数量持续下降，并随时间推移逐步趋于稳定。清除小胶质细胞后，在AOH模型中，RGC存活数量得到显著提高。