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

我们利用大鼠原代海马培养建立了β淀粉样蛋白毒性模型。取18日龄SD胎鼠海马细胞，用无血清培养基培养。我们已经成功地进行了7天的低密度培养，神经突生长良好，没有增加胶质细胞。与对照组相比，暴露于5μ淀粉样蛋白1-42肽（Aβl-42）减少了细胞数量、神经突数量和长度。与暴露于Aβ1-42的细胞相比，在Aβ1-42细胞中添加酮可使细胞存活数量增加一倍，细胞大小和神经突生长增加。尽管我们已经开发了隔膜培养和海马培养，但在这一点上，胆碱能神经元的免疫染色是不稳定的，无法获得定量数据。已知a - β1-42在阿尔茨海默病患者的大脑中增加了6倍，并且a - β1-42通过激活蛋白激酶使间隔神经元中的丙酮酸脱氢酶（PDH）复合物失活。D-(-)-β-羟基丁酸酯（DBHB）是脑内正常的底物，通过绕过PDH络合物直接向TCA循环提供乙酰辅酶a，并产生还原剂NADH。认为dbb对海马神经元a - β1-42毒性的修复机制与上述机制有关。这些数据表明，酮体对a β1-42水平升高具有神经保护作用，这表明酮体可能是治疗阿尔茨海默病和其他与能量危机相关的神经系统疾病的潜在候选者。

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.