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【目的】研究多巴胺诱导小脑颗粒神经元凋亡的分子机制 ,以及胆碱受体激动剂氨甲酰胆碱对多巴胺诱导凋亡的作用。【方法】在培养的小脑颗粒神经元建立多巴胺凋亡模型。用相差显微镜观察形态学 ,DNA凝胶电泳和Hoechst 332 5 8核染色分析神经元凋亡 ,细胞的存活率用二乙酸荧光素 (FDA)染色法检测。采用Westernblot分析细胞外信号调控的蛋白激酶 (ERK)激活情况。【结果】多巴胺可诱导小脑颗粒神经元凋亡 ,并可持续激活ERK ,二者均可被氨甲酰胆碱和PD 980 5 9抑制。氨甲酰胆碱对神经元的保护作用及对ERK激活的抑制作用可被阿托品阻断。【结论】多巴胺在小脑颗粒神经元诱导凋亡可能是通过持续激活ERK介导的。氨甲酰胆碱通过激活M胆碱受体 ,继而抑制了ERK的激活 ,从而起到对神经元的保护作用
【Objective】 To investigate the molecular mechanism of dopamine-induced apoptosis of cerebellar granule neurons and the effect of cholinergic agonist carbachol on dopamine-induced apoptosis. 【Methods】 Dopamine apoptosis model was established in cultured cerebellar granule neurons. Morphological, DNA gel electrophoresis and Hoechst 332 58 nuclear staining were used to observe the apoptosis of neurons. The viability of the cells was detected by FDA staining. Western blot analysis of extracellular signal-regulated protein kinase (ERK) activation. [Results] Dopamine induced apoptosis of cerebellar granule neurons and sustained activation of ERK, both of which were inhibited by carbachol and PD 980 5 9. The protective effect of carbachol on neurons and the inhibition of ERK activation can be blocked by atropine. 【Conclusion】 Dopamine induces apoptosis in cerebellar granule neurons possibly through sustained activation of ERK. Carmcholine protects neurons by activating the M choline receptor, which in turn inhibits the activation of ERK