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背景:淀粉样β蛋白是老年斑的核心成分,其毒性片段淀粉样β蛋白25-35近年广泛用于实验研究中。已往研究表明自制中药复智散可促进培养神经细胞存活,可能具备治疗阿尔茨海默病的效用。目的:研究复智散对抗淀粉样β蛋白25-35对培养神经细胞的毒性作用及其发挥这一作用的可能途径。设计:以细胞为研究对象,重复测量设计。单位:一所大学医院的神经内科和一所大学医院的脑老化重点实验室。材料:实验于2002-06/2003-04在宣武医院北京脑老化重点实验室完成。人多巴胺能神经母细胞瘤株SH-SY5Y细胞。淀粉样β蛋白25-35片段。中药复智散文火煎制,留取上清冷冻抽干配成浓度为0.5g/mL的贮存液备用。抗体:cAMP应答元件结合蛋白、B淋巴细胞白血病-2基因产物中的Bcl-2、细胞色素C由宣武医院北京脑老化研究实验室制备。方法:用不同剂量的淀粉样β蛋白25-35单独或与复智散共同孵育SH-SY5Y细胞,与空白对照组相比,测定在不同孵育条件下培养神经细胞的噻唑蓝代谢率。利用Western-blot法测定复智散单独孵育、淀粉样β蛋白25-35单独孵育及两者共同孵育条件下与空白对照相比蛋白质表达的变化。主要观察指标:各实验组与空白对照组相比的噻唑蓝代谢率。与神经细胞存活及死亡相关蛋白cAMP应答元件结合蛋白,Bcl-2及细胞色素C的表达水
BACKGROUND: Amyloid β protein is the core component of senile plaques. Its toxic fragment amyloid β-protein 25-35 has been widely used in experimental studies in recent years. Previous studies have shown that self-made Fuzhi San can promote the survival of cultured nerve cells and may have the effect of treating Alzheimer’s disease. Objective: To study the toxic effects of Fuzhisan on the protection of cultured neurons against amyloid β-protein 25-35 and possible ways to exert this effect. Design: Repeat the measurement design with cells as the research object. Units: Neurology in a university hospital and a brain aging laboratory at a university hospital. MATERIALS: The experiment was performed at the Beijing Brain Aging Laboratory, Xuanwu Hospital from June 2002 to April 2003. Human dopaminergic neuroblastoma cell line SH-SY5Y cells. Amyloid β-protein 25-35 fragment. Traditional Chinese medicine Fuzhi prose preparation, leaving the supernatant freeze-dried into a stock concentration of 0.5g/mL reserve. Antibodies: The cAMP response element binding protein, Bcl-2 in the B lymphocyte leukemia-2 gene product, and cytochrome C were prepared by the Beijing Brain Aging Research Laboratory of Xuanwu Hospital. METHODS: SH-SY5Y cells were incubated with different doses of amyloid β-protein 25-35 alone or with Fuzhi San. Compared with the blank control group, the metabolic rate of thiazole blue in cultured neurons under different incubation conditions was determined. Western-blot method was used to determine the changes of protein expression of Fuzhisan alone incubated with amyloid β-protein 25-35 alone and in combination with the blank control. MAIN OUTCOME MEASURES: The metabolic rate of thiazole blue in each experimental group compared with the control group. Expressions of cAMP response element-binding protein, Bcl-2 and cytochrome C in neurons