Fundamental research about the ketone body therapy for Alzheimer's disearse.

我们利用原代培养的大鼠海马区建立了淀粉样β蛋白毒性模型。取18天胎龄SD大鼠海马神经细胞，用无血清无血清培养。我们成功地进行了7天以上的低密度培养，神经突起生长良好，没有增加胶质细胞。暴露于5μ淀粉样蛋白S 1-42肽(AβL-42)后，与对照组相比，细胞数、突起数和长度均减少。在暴露于Aβ1-42的细胞中加入酮，与暴露于Aβ1-42的细胞相比，存活的细胞数量增加一倍，细胞大小和突起生长增加。即使我们已经发展了隔膜培养和海马区培养，胆碱能神经元的免疫染色在这一点上仍然不稳定，无法获得定量的数据。众所周知，阿尔茨海默病患者脑中的Aβ1-42增加了6倍，并且Aβ1-42通过激活蛋白激酶来灭活隔神经元丙酮酸脱氢酶复合体。D-(-)-羟基丁酸酯(D-(-)-β-羟基丁酸酯)是脑内一种正常的底物，它绕过丙二醛复合体，直接向三氯乙酸循环提供乙酰辅酶A，并产生还原剂NADH。有研究认为，D-(-)-羟丁酸通过上述机制对A-β1-42所致的海马神经元毒性具有保护作用。这些数据表明，随着Aβ1-42水平的增加，酮体可以提供神经保护作用，表明酮体可能是治疗阿尔茨海默病和其他与能源危机相关的神经疾病的潜在候选者。

We have created the Amyloid beta protein toxicity model using rat primary hippocampal culture. Hippocampal cells are taken from 18 day fetal SD rats, which were cultured with absolute serum free medium. We have succeeded low density culture more that 7 days, with well neurites grouth and without increasing glial cells. Exposure of 5μ amyloicl s 1-42 peptide (Aβl-42) decreased cell number and neurite number and length in comparison to control. Addition of ketones to cells exposed to Aβ1-42 doubled the survived cell number and increased cell size and neurite outgrowth compared with cells exposed to Aβ1-42. Even we have developed the septum culture as well as hippocampal culture, the immunostaining with cholinergic neurons is unstable to have a quantitative data at this point.It is known that Aβ1-42 is increased up to 6 fold in brains from patients with Alzheimer's disease and that Aβ1-42 inactivates the pyruvate dehydrogenase (PDH) complex in septal neurons by activating the protein kinase. D-(-)-β-hydroxybutyrate (DBHB) is a normal substrate in brain, with provides Acetyl CoA to TCA cycle directly with bypassing PDH complex, and also produce the reducing agent NADH.It was thought that DBHB could rescue hippocampal neurons from the toxicity of Aβ1-42 with mechanism mentioned above. These data, which resulted that ketone bodies provide a neuroprotective effect against with increased levels of Aβ1-42, suggest that ketone body could be a potential candidate for use in the treatment of Alzheimer's disease and other neurological diseases associated with energy crisis.