摘要
【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 。
To investigate the molecular mechanism of dopamine (DA) induced apoptosis in cultured cerebellar granule neurons (CGNs) and the effect of muscarinic cholinergic receptor (mAChR) agonist carbachol on it. The apoptosis of neurons was measured by phase contrast microscopy, Hoechst 33258 nucleus staining and DNA fragmentation agarose gel electrophoresis. The neuronal viability was measured by fluorescein diacetate (FDA) staining. The activation of extracellular signal regulated protein kinase (ERK) was determined by Western blot. Dopamine increases the phosphorylation of ERK and induces apoptosis in CGNs, which is blocked by both carbachol and PD 98059. The protective effect and the inhibiting ERK phosphorylation of carbachol were blocked by atropine. [Conclusion] DA induced apoptosis in CGNs may be mediated by activation of ERK. Carbachol protects CGNs from DA induced apoptosis by activating mAChR and subsequent inhibition of activation of ERK.
出处
《中山医科大学学报》
CSCD
北大核心
2001年第3期161-164,169,F002,共6页
Academic Journal of Sun Yat-sen University of Medical Sciences
基金
ThegrantfromtheNationalScientificFoundationfortheOutstandingYoungScientistsofChina ( 396 2 5 0 2 2 )
