二氮嗪预处理对缺氧复氧后大鼠海马神经元凋亡及JNK表达的影响  

Effects of Diazoxide Preconditioning on JNK Expression and Anoxia-Reoxygenation-Induced Apoptosis of Hippocampal Neurons

作　　者：刘荣国[1] 崔翔[1] 宋娜[1] 陈彦青[1] 

机构地区：[1]福建医科大学省立临床医学院福建省立医院麻醉科,福州350001

出　　处：《福建医科大学学报》2010年第1期24-27,共4页Journal of Fujian Medical University

基　　金：福建省自然科学基金(2006J0083)

摘　　要：目的探讨二氮嗪(DZ)预处理能否抑制C-Jun氨基末端激酶(JNK)表达而减轻缺氧复氧后大鼠海马神经元的凋亡。方法原代培养9～10 d的SD大鼠海马神经元随机分为5组:对照组、DZ 0,30,100μmol/L和DZ 100μmol/L+5-羟癸酸(5-HD)100μmol/L预处理组。除对照组外,其余4组神经元自缺氧前2 d开始,每天实施以上对应浓度DZ预处理1 h,连续3 d。于体外缺氧4 h复氧48 h后,采用四唑蓝比色法测定海马神经元活力,AnnexinⅤ-FITC流式细胞术测定凋亡率,Western blotting法检测JNK蛋白表达。结果与对照组比较,缺氧复氧后海马神经元的活力下降,凋亡率增大,JNK蛋白表达增强(P<0.05)。与其他预处理组比较,DZ100μmol/L预处理组的神经元活力高,凋亡率低(P<0.05),其他预处理组间比较无统计学意义。DZ预处理后,JNK蛋白表达减低,且DZ 100μmol/L组表达弱于DZ 30μmol/L组(P<0.05)。同时给于5-HD预处理后,DZ未能抑制JNK蛋白表达。结论DZ预处理可能抑制JNK表达而减轻缺氧复氧后大鼠海马神经元的凋亡。Objective To study the effect of diazoxide （DZ） preconditioning on JNK protein expression and on anoxia-reoxygenation-induced apoptosis of hippocampal neurons. Methods Cultured for 9-10 d in vitro, the hippocampal neurons of SD rats were divided into the following 5 groups randomly： Control, DZ 0μmol/L, DZ 30 μmol/L, DZ 100 μmol/L, and DZ 100 μmol/Lq- 5-hydroxydeeanoate 100μmol/L （5-HD, a selective MitOKATP channel blocker）. Prior to oxygen deprivation, the hippocampal neurons except those in the control group were treated with the above concentrations of DZ or DZ＋5- HD for 1 h per day and for 3 d. Thereafter, neurons were subjected to anoxia for 4 h and followed by reoxygenation. After 48 h of reoxygenation, the neuronal vitality was measured by MTT method. The apoptotic rates were assayed by annexinV-FITC staining. The expression of JNK was detected and eval- uated by western blotting. Results Anoxia-reoxygenation injury reduced the neuronal vitality and increased the apoptotie rates significantly. In comparison with other anoxic groups, the DZ 100 μmol/L group showed an increased vitality but simultaneously decreased apoptotic rate（P〈0.05）. 5-HD could a- bolish the neuroprotection provided by 100μmol/L DZ. JNK in control normoxic neurons was expressed slightly while anoxia-reoxygenation led to its high expression. The administration of DZ reduced the expression of JNK significantly （P〈0.05）. Compared with DZ 30 μmol/L, DZ 100 μmol/L further decreased the expression of JNK protein （P〈0.05）. However, effects of DZ on the expression of JNK were reversed after the co-treatment with 5-HD. Conclusions DZ alleviates the anoxia-reoxygenation-in- duced apoptosis of hippoeampal neurons possibly through down-regulating the expression of JNK protein.

关 键 词：二氮嗪 海马 疾病模型 动物 缺氧缺血 脑 神经元 细胞凋亡 JNK丝裂原活化蛋白激酶类 

分 类 号：R96[医药卫生—药理学]