二氮嗪预处理联合低温对大鼠海马神经元缺氧复氧时Bcl-2和Bax表达的影响  

Effects of diazoxide preconditioning combined with hypothermia on expression of Bcl-2 and Bax during anoxia-reoxygenation in rat hippocampal neurons

作　　者：刘荣国[1] 陈彦青[1] 崔翔[1] 宋娜[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,福州市350001

出　　处：《中华麻醉学杂志》2009年第6期543-546,共4页Chinese Journal of Anesthesiology

基　　金：福建省自然科学基金资助项目（2006J0083）

摘　　要：目的二氮嗪预处理联合低温对大鼠海马神经元缺氧复氧时Bcl-2和Bax表达的影响。方法清洁级SD大鼠，出生〈24h，体重5～6g，离体培养海马神经元，接种于培养皿或96孔板。海马神经元随机分为8组，每组36孔或12皿：常温组（NT组）、二氮嗪预处理＋常温组（DP＋NT组）、浅低温组（MiH组）、二氮嗪预处理＋浅低温组（DP＋MiH组）、中低温组（MoH组）、二氮嗪预处理＋中低温组（DP＋MoH组）、深低温组（DeH组）和二氮嗪预处理＋深低温组（DP＋DeH组）。DP＋NT组、DP＋MiH组、DP＋MoH组和DP＋DeH组培养液中加入二氮嗪，终浓度为100μmol／L，孵育1h，1次／d，连续2d，随后分别在37、34、30和22℃下缺氧4h，37℃复氧48h。NT组、MiH组、MoH组和DeH组分别在37、34、30和22℃下行缺氧4h，37℃复氧48h。于复氧48h时测定海马神经元活力、凋亡率和Bcl-2和Bax的表达水平，计算Bcl-2／Bax。结果与NT组比较，DP＋NT组、MiH组、MoH组和DeH组海马神经元活力升高，凋亡率降低，Bcl-2表达上调，Bax表达下调，Bcl-2／Bax升高（P〈0．05）；与DP＋NT组比较，DP＋MiH组、DP＋MoH组和DP＋DeH组海马神经元活力升高，早期凋亡率降低，Bcl-2表达上调，Bax表达下调，Bcl-2／Bax升高（P〈0．05），晚期凋亡率差异无统计学意义（P〉0．05）；与MiH组比较，DP＋MiH组和DeH组海马神经元活力升高，早期凋亡率降低，Bcl-2表达上调，Bax表达下调，Bcl-2／Bax升高（P〈0．05），晚期凋亡率差异无统计学意义（P〉0．05），MoH组上述指标差异无统计学意义（P〉0．05）；与DP＋MiH组比较，DP＋DeH组海马神经元活力升高，早期凋亡率降低，Bcl-2表达上调，Bax表达下调，Bcl-2／Bax升高（P〈0．05）。结论二氮嗪预处理联合低温减轻大鼠神经元缺氧复氧损伤的机制可能与纠正Bcl-2与Bax失衡，抑制神经元早期凋亡有关。Objective To investigate the effects of diazoxide preconditioning combined with hypothermia on the expression of Bcl-2 and Bax during anoxia-reoxygenation in rat hippocampal neurons. Methods The hippocampal neurons isolated from newborn SD rats （ 〈 24 h, weghing 5-6 g） were inoculated in the culture dish or 96 well plates. The hippocampal neurons were randomly assigned into 8 groups and each group contained 36 wells or 12 dishes of neurons： normal temperature group （group NT）, diazoxide preconditioning （DP） ＋ NT group （group DP＋ NT）, mild hypothermia group （group MiH）, DP ＋ MiH group （group DP ＋ MiH）, moderate hypothennia group （group MoH）, DP ＋ MoH group （ group DP ＋ MoH）, deep hypothermia group （ group DeH） and DP ＋ DeH group （ group DP ＋ DeH）. In group DP ＋ NT, DP ＋ MiH, DP ＋ MoH and DP ＋ DeH, diazoxide was added to the culture media, the final concentration was 100 μmol/L, and the neurons were incubated for 1h once a day for 2 d, and then subjected to 4 h of bypoxia at 37, 34, 30 and 22℃, respectively, followed by 48 h of reoxygenation at 37℃ . Tile neuronal viability, apoptotic rates and expression of Bcl-2 and Bax were determined and the ratio of Bcl-2/Bax was calculated. Results The neuronal viability was significantly higher, apoptotic rate lower, Bcl-2 expression higher,Bax expression lower and Bcl-2/Bax ratio higher in group DP ＋ NT, MiH, MoH and DeH than in group NT （P 〈 0.05）. The neuronal viability was significantly higher, early apoptotic rate lower, Bcl-2 expression higher, Bax expression lower and Bcl-2/Bax ratio higher in group DP ＋ MiH, DP ＋ MoH and DP ＋ DeH than in group DP ＋ NT （ P 〈 0.05 ）, but there was no significant difference in the late apoptotic rate between group DP ＋ MiH, DP ＋ MoH, DP ＋ DeH and DP ＋ NT （P 〉 0.05）. The neuronal viability was significantly higher, early apoptotic rate lower, Bcl-2 expression higher, Bax expression lower and Bcl-2/Bax ratio higher in group DP ＋

关 键 词：二氮嗪 缺血预处理 低温 原癌基因蛋白质C-BCL-2 Bcl-2相关X蛋白 再灌注损伤 海马 

分 类 号：R741[医药卫生—神经病学与精神病学]