新型的治疗阿尔茨海默氏症药物(1-二甲基磷酰基-2，2，2-三氯)-乙基-1-醇烟酸酯对体外神经细胞的作用

Effects of a new drug-NMF for Alzheimer's disease on neurons in vitro

目的：观察本实验室合成的一种治疗阿尔茨海默氏症(AD)的药物(1-二甲基磷酰基-2，2，2-三氯)-乙基-1-醇烟酸酯(NMF)，对体外培养的皮层神经细胞活性的影响以及对海人藻酸(KA)所致的神经损伤的保护作用。方法：采用体外培养皮层神经元的方法，解剖分离15d胚胎小鼠皮层神经细胞，接种于96孔板，48h后加药并培养72h，以MTT法观察NMF对小鼠皮层神经细胞活性的影响；同时将接种于24孔板的细胞预先给予NMF，d3时加或不加KA处理后，以台盼蓝染色鉴别并计数死、活细胞，可得出细胞的存活率。结果：NMF明显促进胎鼠皮层神经元活性，其中NMF 1、0．1、10 nmol·L^(-1)促进神经元活性增殖率分别高达34．7％、37．4％、36．7％。NMF明显促进正常胎鼠皮层神经元存活率，与对照组比较，10、1nmol·L^(-1)NMF对皮层神经元的存活率分别提高39．3％、73．5％。NMF能显著对抗KA所致的神经元损伤，与KA损伤组相比，NMF0．1、10、1nmol·L^(-1)对损伤皮层神经元的保护率分别为77．30％、80．10％、84．15％。结论：NMF明显促进胎鼠皮层神经元的活性、提高正常皮层神经元的存活率，并能有效地保护KA所致的神经元损伤，提示NMF是一种很有潜力的治疗AD的药物。

AIM： To observe the effects of NMF, a drug synthesized for Alzheimer＇s Disease in our laboratory, on viability of mouse cortical neurons and provide protection against the excitotoxic effect of kainic acid （KA） on primary cortical cultures. METHODS： Primary cultured neurons were dissociated from 15 d old mouse embryo and plated on 96-well culture-plates for 48 h, then NMF was added in medium for 72 h, and MTF method was used to determine the effect of NMF on neuron viability. At the same time, neurons plated on 24-well culture-plates with NMF in the medium treated with or without KA on d 3. Dying cells were identified by trypan blue under phase contrast microscope and survival rate came up. RESULTS： NMF increased the viability of primary neurons, especially on the dose of 0.1,1μmol·L^-1 , 10 nmol·L^-1 and the rate enhanced was 34.7%, 37.4% and 36.7 %, respectively; culturing cortical neurons with NMF at 1, and 10 nmol·L^-1 for 4 d increase the survival of neurons at 39.3%, 73.5% and decreased the death rate of cells. Damaged neurons induced by cultured with KA for 30 rain showed various changes and higher mortality. While pretreated with NMF 0.1, 10 nmol·L^-1, 1nmol·L^-1 for 4 d, the cortical neurons were resistant to the injury of KA and the protective rate were 77.30%, 80.10%, and 84.15%, respectively. CONCLUSION： NMF can increase the viability and survival rate of mouse cortical neurons, and decrease the mortality and protect neurons against KA induced neurotoxicity.