一氧化氮供体硝普钠对海马神经元cpp32基因表达的影响(英文)  

Influence of the nitric oxide-donor of sodium nitroprusside on the expression of hippocampal neurons gene cpp32

作　　者：刘勇军[1] 张海风[1] 祝其锋[1] 梁爱玲[1] 

机构地区：[1]广东医学院生物化学与分子生物学研究所,广东省湛江市524023

出　　处：《中国临床康复》2006年第14期182-184,F0003,共4页Chinese Journal of Clinical Rehabilitation

基　　金：广东省自然科学基金资助项目(04011397,5011588);广东省医学科研基金资助项目(A2003544);广东省教育厅重点扶持学科(粤教科[2004]1号)~~

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。BACKGROUND： Alzheimer disease is a senile degenerative disease of nervous system. Neuron apeptosis 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, bat 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.

关 键 词：硝普钠 海马 神经元 基因表达 

分 类 号：R745.1[医药卫生—神经病学与精神病学]