氨基胍对氧-糖剥夺大鼠皮质神经元一氧化氮及半胱氨酸天冬氨酸蛋白酶3表达的影响研究  

Effect of Aminoguanidine on the Expression of NO and Caspase-3 in Rat Cortical Neurons After Oygen Glucose Deprivation

作　　者：唐仕军[1] 赵冬[1] 朱立仓[1] 李晓天[1] 朱文学[1] 杨鹏[1] 王业忠[1] 

机构地区：[1]石河子大学医学院第一附属医院神经外科,新疆石河子市832000

出　　处：《中国全科医学》2016年第12期1429-1434,共6页Chinese General Practice

基　　金：国家自然科学资金资助项目(81360185);新疆生产建设兵团博士基金资助项目(2011BB016)

摘　　要：目的探讨氨基胍对氧-糖剥夺大鼠皮质神经元一氧化氮(NO)及半胱氨酸天冬氨酸蛋白酶3(Caspase-3)表达的影响。方法 2014年11月—2015年7月选取新生24 h内SD大鼠12只,分离及培养大鼠皮质神经元,取原代培养8 d的皮质神经元,分为3组:正常对照组、氧-糖剥夺组、氨基胍组。正常对照组皮质神经元采用Neurobasal培养液+2%B27培养液添加剂培养,氧-糖剥夺组皮质神经元缺氧、缺糖处理45 min后,换Neurobasal培养液+2%B27培养液添加剂培养;氨基胍组皮质神经元缺氧、缺糖处理45 min后,换Neurobasal培养液+2%B27培养液添加剂培养的同时加入终浓度为10 mmol/L的氨基胍溶液。各组培养2、6、12 h,采用硝酸还原法检测细胞上清液中NO表达水平,采用Western blotting法检测Caspase-3表达水平。结果各组培养2、6、12 h时细胞上清液中NO、Caspase-3表达水平比较,差异均有统计学意义(P<0.05);其中氧-糖剥夺组、氨基胍组不同时间点细胞上清液中NO、Caspase-3表达水平较正常对照组升高(P<0.05);氨基胍组不同时间点细胞上清液中NO、Caspase-3表达水平较氧-糖剥夺组降低(P<0.05)。结论氨基胍可抑制氧-糖剥夺大鼠皮质神经元NO及神经元凋亡相关基因Caspase-3表达水平,从而抑制皮质神经元的凋亡,对皮质神经元具有保护作用。Objective To investigate the effect of aminoguanidine on the expression of nitric oxide（NO）and Caspase- 3 in rat cortical neurons after oxygen and glucose deprivation. Methods From December 2014 to July 2015,12 SD rats born within 24 hours were selected. The cortical neurons of the rats were separated and cultured,and cortical neurons of primary culture for 8 days were obtained and were divided into three groups：normal control group,oxygen - glucose deprivation group and aminoguanidine group. The cortical neurons of normal control group were cultured by Neurobasal ＋ 2% B27 nutrient solution;the cortical neurons of oxygen - glucose deprivation group received 45 min management for hypoxia and lack of sugar,and converted to be cultured by Neurobasal ＋ 2% B27 nutrient solution;the cortical neurons of aminoguanidine group received 45 min management for hypoxia and lack of sugar,and converted to be cultured by Neurobasal ＋ 2% B27 nutrient solution plus 10 mmol/ L aminoguanidine solution. After culture for 2,6 and 12 h,the NO expression in cell supernatant was detected using nitric oxide assay kit,and Caspase - 3 expression was detected using Western blotting method. Results The three groups were significantly different in the expression levels of NO in cell supernatant and Caspase - 3 at 2,6 and 12 h（P 〈 0. 05）. Oxygen -glucose deprivation group and aminoguanidine group were higher than normal control group in the expression levels of NO in cell supernatant and Caspase - 3 at different time points（ P 〈 0. 05）. Aminoguanidine group was lower than oxygen - glucose deprivation group in the expression levels of NO in cell supernatant and Caspase - 3 at different time points（ P 〈 0. 05）. Conclusion Aminoguanidine can inhibit the expression of NO and apoptosis related gene Caspase - 3 in the rat cortical neurons after oxygen and glucose deprivation,which inhibits the apoptosis of cortical neurons and has a protective effect on the neurons.

关 键 词：神经元 一氧化氮 半胱氨酸天冬氨酸蛋白酶3 氨基胍 

分 类 号：R743.33[医药卫生—神经病学与精神病学]