【摘要】 背景:阿尔茨海默病是一种老年神经系统退行性疾病,神经元凋亡被认为是其可能的发病原因之一,体外细胞培养是研究其凋亡机制的常用方法。目的:观察一氧化氮供体硝普钠对体外培养的海马神经元cpp32基因表达的影响。设计:随机对照动物实验。单位:广东医学院生物化学与分子生物学研究所。材料:实验在广东医学院生物化学与分子生物学研究所完成,实验动物为新生(出生24h以内)SD大鼠。方法:取大鼠海马神经元进行原代培养,采用终浓度分别为0,25,50,100,200,400μmol/L的硝普钠处理海马神经元24h,用反转录聚合酶链反应检测mRNA表达变化,Westernblot检测蛋白表达的变化;再用终浓度分别为0,25,50,100,200μmol/L的硝普钠处理海马神经元12h,用CPP32活性检测试剂盒检测CPP32酶活性。主要观察指标:cpp32mRNA表达、CPP32蛋白表达及CPP32酶活性检测。结果:随着硝普钠剂量的增加,cpp32mRNA表达无改变;但CPP32酶原被裂解活化,从硝普钠50μmol/L开始,酶活性显著增加,为对照组的3.02倍,100μmol/L达最大值,为对照组的3.47倍。结论:硝普钠不增加cpp32mRNA的表达,但可诱导CPP32酶原的裂解,使CPP32活化。更多还原

【Abstract】 BACKGROUND Alzheimer disease is a senile degenerative disease of nervous system. Neuron apoptosis is regarded as one of possible reasons and neuron culture in vitro is a common method to research the mechanism of apoptosis. OBJECTIVE To observe the influences of nitric oxide-donor sodium nitroprusside SNP on the expression of gene cpp32 in the cultured hippocampal neurons in vitro. DESIGN A randomized controlled animal experiment. SETTING Institute of Biochemistry and Molecular Biology in Guangdong Medical College. MATERIALS The experiment was carried out in the Institute of Biochemistry and Molecular Biology Guangdong Medical College and newborn < 24 hours Sprague-Dawley rats were used. METHODS The hippocampl neurons of rats were primarily cultured and then treated with SNP of different terminal concentrations 0 25 50 100 200 400 μmol/L for 24 hours and the expressions of mRNA and protein were analyzed with RT-PCR and Western blot respectively. The hippocampl neurons of rats were treated with SNP of different terminal concentrations 0 25 50 100 200 400 μmol/L for 12 hours and the activity of CPP32 enzyme was detected with CPP32 activity detected kit. MAIN OUTCOME MEASURES The expressions cpp32 mRNA and CPP32 protein and activity of CPP32 were detected. RESULTS The cpp32 mRNA expression was unchanged as the increasing dose of SNP but the pro-CPP32 was activated and the activity of CPP32 was increased significantly at 50 μmol/L SNP which was 3.02 times of that in the control group and reached to the maximal value at 100 μmol/L which was 3.47 times of that in the control group. CONCLUSION SNP cannot increase the cpp32 mRNA expression but can increase degradation of pro-CPP32 and activate CPP32.更多还原