β淀粉样蛋白对小胶质细胞合成一氧化氮影响.docVIP

β淀粉样蛋白对小胶质细胞合成一氧化氮影响 　 作者：韩笑峰 吕丽 葛汝丽 唐荣华 【摘要】 目的 探讨β淀粉样蛋白(amyloid betapeptide, Aβ)诱导小胶质细胞活化后κ轻链核因子(nuclear factorkappa B, NFκB)的表达及一氧化氮(NO)水平的变化。方法 Aβ干预纯化培养的小胶质细胞，观察其形态学变化，采用镉还原法测定NO水平，免疫细胞化学方法研究NFκB的表达。结果 500 nmol/L及1000 nmol/L Aβ干预组细胞形态呈“阿米巴样”，细胞核内NFκB的表达增加（P＜0.05），培养基中NO浓度升高（P＜0.05）。结论 Aβ激活小胶质细胞NFκB途径大量合成NO，可能参与阿尔茨海默病(Alzheimers disease, AD)的致病过程。 【关键词】 β淀粉样蛋白 小胶质细胞 κ轻链核因子 一氧化氮 阿尔茨海默病 【Abstract】 Objective To study the morphological changes of microglial cell activated by amyloid betapeptide, the expression of nuclear factorkappa B increased in nucleus and the levels of nitric oxide in cultural medium. Methods Purified microglial cells were intervened by amyloid betapeptide and the morphological changes were observed . The cadmiumreduction method was used to determine the levels of nitric oxide in culture medium, immunocytochemical method to investigate changes of nuclear factorkappa B. Results There were no statistical significant differences between the experimental group of 100 nmol/L amyloid betapeptide and the control group (P＞0.05). In the experimental groups of 500 nmol/L and 1000 nmol/L amyloid betapeptide, microglial cells were bigger, the expression of nuclear factorkappa B increased in nucleus and the levels of nitric oxide were higher in culture medium than control group (P＜0.05).Conclusion The amyloid betapeptide could participate in the processes of Alzheimer’s disease by inducing microglial cells to excrete nitric oxide. 【Key words】 amyloid betapeptide，microglial cell，nuclear factorkappa B，nitric oxide，Alzheimers disease 阿尔茨海默病(Alzheimers disease, AD)重要的病理特征之一是纤维状β淀粉样蛋白(amyloid betapeptide, Aβ)在神经细胞外及脑血管壁沉积形成老年斑(senile plaques, SP)。大量研究表明，Aβ的沉积在AD发病中起着关键作用，但其神经损伤的具体机制尚不完全清楚，研究提示，一氧化氮(nitric oxide, NO)可能与Aβ的神经毒性有关。本研究用Aβ干预小胶质细胞，观测κ轻链核因子(nuclear factorkappa B, NFκB)的表达以及NO水平的变化，探讨Aβ诱导小胶质细胞合成NO的机制。 1 材料和方法 1.1 材料 1.1.1 动物：出生3 d内的Wistar大鼠，由华中科技大学同济医学院实验动物中心提供。 1.1.2 试剂：DMEM/F12培养基、胎牛血清（购自Hyclone公司），Aβ140、Aβ142