银杏酮酯对β-淀粉样蛋白诱导大鼠海马神经元氧化应激致细胞损伤影响的实验研究  

Ginkgo biloba Extract 50 Inhibited Beta-amyloid-induced Oxidative Stress in Rats′ Hippocampal Neurons:an Experimental Study

作　　者：夏趁意[1] 董献文[1] 赵妍[1] 徐颖[1] 郝莉[1] 张志雄[1] 

机构地区：[1]上海中医药大学生理教研室,上海201203

出　　处：《中国中西医结合杂志》2014年第7期833-838,共6页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：上海市教委重点学科资助项目(No.J50301);上海中医药大学预算内资助项目(No.2013JW02)

摘　　要：目的研究银杏酮酯(GBE50)对由β-淀粉样蛋白(Aβ)诱导的海马神经元氧化应激反应的影响及可能机制。方法通过体外原代培养获得海马神经细胞,将细胞分成4组:正常对照组(Ctrl组)、Aβ组、溶剂对照组(PDO组)和GBE50组(分6个浓度梯度5、10、25、50、100、200μg/mL)。除Ctrl组外,其他组中的细胞均经20μmol/L Aβ25-35诱导后出现氧化应激状态,采用MTT和荧光探针标记法,观察不同浓度的GBE50对海马神经元细胞活力和细胞内活性氧(ROS)的影响。运用Western blotting法检测观察GBE50(25、50、100μg/mL)3个浓度梯度对氧化应激通路中的胞质内细胞色素C(Cyto C)与胞内总Cyto C蛋白表达量之间的比值,以及活化的Caspase-3蛋白表达量的影响。结果与Ctrl组比较,20μmol/L的Aβ25-35诱导细胞后使细胞活力明显降低,细胞内ROS量明显增多(均P<0.05)。与Aβ组比较,GBE50能够改善细胞活力,除10μg/mL浓度外,GBE50其他浓度组的细胞活力随着浓度的增高而明显升高(P<0.05);同时GBE50各浓度组的胞内ROS量均明显减少(均P<0.05)。Aβ诱导后细胞内的Cyto C比值及活化Caspase-3表达量均较Ctrl组明显升高(P<0.05)。与Aβ组比较,在GBE50的3个浓度组中,100μg/mL的GBE50组中Cyto C比值明显降低(P<0.05),50、100μg/mL的GBE50组中活化Caspase-3表达量均显著降低(P<0.05)。结论 20μmol/L的Aβ25-35能够诱导海马神经元胞内活性氧的产生,而GBE50通过减小胞质内与胞内总Cyto C量的比值,抑制凋亡蛋白Caspase-3活化,从而抑制Aβ诱导的细胞内氧化应激反应。Objective To study the in vitro effect and mechanism of Ginkgo biloba Extract 50 （GBE50） for inhibiting beta-amyloid （Aβ）-induced oxidative stress in rats＇ hippocampal neurons. Methods The primary hippocampal neurons were cultured in vitro and divided into 4 groups, i. e. the normal control group （Ctrl）, the Aβ group, the propanediol control group （PDO）, and the six GBE50 concentrations groups （5, 10,25, 50, 100, and 200 μg/mL）. Excepted the Ctrl group, neurons were induced to oxidative stress by 20 μmol/L Aβ25-35.The MTT and fluorescent probes labeling were used to observe the effect of GBE50 with different concentrations on the cell viability and the generation of intracellular reactive oxygen species （ROS） in neurons. Furthermore, Western blot was used to detect the cytoplasmic/total cytochrome C （Cyto C） ratio and total intracytoplasmal Cyto C, and the effect of the expression of oxidative stress-related protein Cyto C and activated Caspase-3 in three GBE50 concentrations groups （25, 50, and 100μg/mL）. Results Compared with the Ctrl group, the cell vitality was obviously lowered and intracellular ROS generation significantly increased after induction of 20 μmol/L Aβ25-35 （both P 〈 0.05）. Compared with the Aβ group, the cell vitality was evidently improved after treated with different GBE50 doses. Except for 10 μg/mL, the cell vitality could be obviously elevated along with increased drug concentrations （P 〈0.05）. Meanwhile, the intracellular ROS generation decreased significantly in each GBE50 dose groups （P 〈0.05）. Aβ could increase the cytoplasmic/total Cyto C ratio and enhance the activated Caspase-3 expression significantly （P 〈0.05）. Compared with the Aβ group, among the three concentrations of GBE50, the Cyto C ratio was obviously lowered in the 100 μg/mL GBE50 group （P 〈 0.05）, and the expression of activated Caspase-3 significantly decreased in 50 μg/mL and 100 p.g/mL GBE50 groups （P 〈0.05）. Conclusions 20 μmol/L Aβ2

关 键 词：银杏酮酯 Β-淀粉样蛋白 海马神经元 氧化应激 

分 类 号：R285.5[医药卫生—中药学]