激活小胶质细胞α_7-nAChR减少Aβ蛋白所致的神经元毒性的研究  被引量:2

Study on activation microglia's α_7- n AChR for reducing neuron toxicity induced by Aβ protein

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作  者:雷露雯[1] 李伟煊[1] 冯智敏[1] 邓冲[1] 陈翔[1] 

机构地区:[1]南方医科大学附属南海区人民医院,广东佛山528000

出  处:《药学研究》2015年第8期435-438,共4页Journal of Pharmaceutical Research

基  金:广东省自然科学基金资助项目(No.07005203)

摘  要:目的研究激活小胶质细胞上的α7-烟碱型乙酰胆碱受体对减少由于β-淀粉样蛋白1-42沉积所致的慢性炎性反应对神经元毒性作用。方法取孕18 d的胎鼠大脑皮质做原代神经元的培养,选用微管相关蛋白-2与神经元特异性烯醇化酶作为神经元特异性标志物,用免疫细胞化学技术对神经元进行鉴定。培养10 d的神经元与小胶质细胞共同培养于带有膜插件的培养皿中。试验分空白对照组、β-淀粉样蛋白组、共培养组、烟碱预处理组,各组添加β-淀粉样蛋白1-42共培养96 h后,用流式细胞仪及免疫荧光法检测神经元的凋亡率。结果体外培养10 d的神经元,用免疫荧光及DAB显色鉴定神经元均能显色,细胞纯度达到了94.49%。激活组与未激活组相比神经元的凋亡率明显下降(P<0.05)。结论激活小胶质细胞上的α7-烟碱型乙酰胆碱受体能减少β-淀粉样蛋白介导的神经元凋亡,对神经元产生保护作用。Objective To investigate the activation of microglia’ s α7neuromal acetylcholine receptor( α7- n AChR)for reducing neuron toxicity,which resulted from accumultation of β- amyloid peptide 1- 42( Aβ1- 42) increased levels of pro- inflammatory factors,contributed to the local jnflammatory response. Methods The cortex of fetation 18 days rats were isolated and minced. The growth of neurons was observed with contrast phase miroscop. The cells were identified by immunocytochemistry after immunostaining with MAP- 2 and NSE on the tenth day. The tenth day neurons coculture with microglai cells in the transwells. The cells randomly divided into four groups: blank control,Aβ protein group,coculture group;nicotin pretreat group. Neurons apoptosis model was established by added Aβ1- 42 protein in each group after 96 hours,and the apoptosis rats was analysed by FCM and fluorescence microscopy. Results Neuron cells after 10 days incubation were identified by immunocytochemistry,immunofluorescence staining and DAB staining show neuron cells stainning positive,purity ratio reached 94. 49%. The neuron apoptosis ratio in activative microglia’s α7- n AChR group was lower than that in inactivative group. Conclusion Activative microglia’s α7- n AChR reduced neuron apoptosis ratio,so that protected neuron from the toxic effect induced by pro- inflammatory factors.

关 键 词:α7-烟碱型乙酰胆碱受体 神经元 免疫细胞化学 细胞凋亡 

分 类 号:R965[医药卫生—药理学]

 

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