摘要
目的 探讨奥美拉唑（OME）对缺氧缺糖（OGD）诱导的神经元损伤的作用及可能机制.方法选取妊娠15~16 d的SD孕鼠1只,提取新生大鼠大脑皮质神经元体外培养.7 d后将培养的神经元随机分为正常对照组（Control组）、模型组（OGD组）和OGD＋OME组（分别加入3、10、30、100、300 μg/ml的OME）,采用活细胞计数法检测神经元的增殖能力,Hoechst 33342荧光染色及TUNEL法观察神经元损伤和凋亡,Annexin V/PI双染流式细胞术检测神经元凋亡率;观察ALDH2激动剂Adal1和阻滞剂Cya对OGD神经元凋亡的影响;收集Control组、OGD组和OGD＋OME组（10 μg/ml的OME）细胞,测定乙醛脱氢酶2（ALDH2）活性.结果 与Control组比较,OGD使体外培养的原代神经元增殖能力下降[OD450：（0.370±0.027）与（0.959±0.062）,P＜0.05],凋亡率增加[（33.53±1.41）%与（15.42±1.29）%,P＜0.05];与OGD组比较,10、30 μg/ ml的OME可促进神经元增殖（P＜0.05）,降低神经元凋亡率（P＜0.05）.与OGD组比较,ALDH2激动剂Adal1能降低神经元凋亡率[（28.92±0.12）%与（16.73±0.62）%,P＜0.05],而其阻滞剂Cya则增加神经元凋亡率[（28.92±0.12）%与（45.22±3.23）%,P＜0.05].与Control组比较,OGD组神经元ALDH2相对活性降低了74.01%,而10 μg/ ml OME使OGD神经元ALDH2相对活性增加了130.00%.结论 OME对氧糖剥夺神经元具有保护作用,这可能与抗凋亡、激活ALDH2活性有关.
Objective To investigate the effects of omeprazole （OME） on injured neuron induced by oxygen glucose deprivation （OGD） and the possible mechanism. Methods A SD pregnant rats was selected, on day 15 - 16 of gestation, neu- rons isolated from newborn rat cerebral cortex were cultured in vitro for seven days, then neurons were randomly divided into con- trol group, OGD group, and OGD ±OME group （3, 10, 30, 100, 300 μg/ml OME was added in separately） . Proliferation of neurons was detected by CCK - 8. Injury and apoptosis of neurons were detected by Hoechst 33342 fluorescence staining and TUNEL, neurons apoptosis rate was detected by Annexin V/PI flow eytometry. To observe the effects of ALDH2 agonist （ Adall ） and retardant （Cya） on injured neuron induced by OGD. Neurons of control group, OGD group and OGD ± OME group （10 μg/ ml OME） were collected, the activities of ALDH2 were measured. Results Proliferation of primary neurons which were cultured in vitro in OGD group was significantly lower than that in control group [ OD450 ： （0. 370 ± 0. 027 ） vs. （0. 959 ± 0. 062）, P 〈 0.05]. Apoptosis rate of primary neurons in OGD group was significantly higher than that in control group [ （33.53 ± 1. 41 ） % vs. （ 15.42 ± 1.29 ） % , P 〈 0. 05] . Proliferation of primary neurons in OGD ± OME group （ 10, 30 μg/ml OME） was signifi- cantly higher than that in OGD group （P 〈0. 05）, apoptosis rate of primary neurons in OGD ± OME group （ 10, 30 μg/ ml OME） was significantly lower than that in OGD group （ P 〈 0. 05 ） . Apoptosis rate of primary neurons in which ALDH2 agonist （Adall） were added was significantly lower than that in OGD group [ （16. 73 ±0. 62）%vs. （28. 92 ±0. 12）%, P〈0. 05]. Apoptosis rate of primary neurons in which retardant （Cya） were added was significantly higher than that in OGD group [ （45.22 ± 3.23）% vs. （28. 92 ± 0. 12）%, P 〈0. 05] . Compared with the control group, the activities of ALDH2 in neurons am
出处
《中国全科医学》
CAS
CSCD
北大核心
2014年第6期636-640,共5页
Chinese General Practice
基金
无锡市科技支撑项目指令性课题(CSE01N1236)
无锡市科技支撑项目指导性课题(CSZ00N1233)
