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-醇烟酸醋对体外神经细胞的作用

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-醇烟酸醋(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 15 d胚胎小鼠皮层神经细胞, 接种于 96孔板,48 h 后加药并培养 72 h,以 MIT 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,d 3 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 NMF1、0. 1、10nmol·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卒,与对照组比较,10nmol·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、10nmol·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洁性、提高正常皮层神经元,的存活卒,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。

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

AIM: To observe the effects of NMP, adrug synthesized for Alheimer's Disease in our laboratory, on viability of mouse cortical neurons and provide pro -tection against the excitotoxic effect of kainic acid ( KA )on primary cortical cultures.METHODS: Primary cultured neurons were dissociated from 15 d old mouse em -bryo and plated on 96-well culture-plates for 48 NMF was added in medium for 72 h, and MTT methodwas 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 blueunder phase contrast microscope and survival rate carneup.RESULTS: NMF increased the viabilityneurons, especially on the dose of 0.1, 1μmol·L^-1, 10nmol·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 4d increase the survivalof neurons at 39.3%, 0.73.5% and decreased the deathrate of cells. Damaged neurons induced by cultured with KA for 30 min showed various changes and higher mortali While pretreated with NMF 0.1, 10nmol·L^-1, 10 nmol·L^-1 for 4d. the cortical neurons were resistant tothe 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 mousecortical neurons, and decrease the mortality and protectneurons against KA induced neurotoxicity.