【作者】 曾杰；

【导师】 雷德亮；

【作者基本信息】 中南大学 ， 神经生物学， 2010， 硕士

【摘要】 目的：探讨17α雌二醇(E2α)对稳定表达人APP/PS1基因的神经母细胞瘤细胞株(Neuro-2a) (APP/PS1 N2a)β-淀粉样蛋白(Aβ)产生的影响及可能机制。方法：将APP/PS1N2a细胞用10 nM E2α预处理2天,更换培养液后,分别进行如下处理：(1)加入不同浓度E2α(25nM,50nM,100nM,200nM)和溶媒(对照组),用Western-blot检测APP/PS1N2a细胞内和细胞外的Ap蛋白水平及细胞BACE1蛋白水平；(2)加入100 nM E2α、20 mM葡萄糖和5 mU葡萄糖氧化酶(glucose oxidase, GOX),在培养液中GOX可以分解G并产生H202,对细胞造成氧化损伤。24h后,进行DHE染色观察APP/PS1 N2a细胞中活性氧(ROS)含量的变化。结果：不同浓度E2α(25nM,50nM,100nM,200nM)处理后,细胞外Ap蛋白水平明显减少,差异具有统计学意义(P<0.05)；而细胞内Ap蛋白水平无明显变化(P>0.05)；细胞BACE1的表达水平无明显变化(P>0.05)；E2α可以减少G/GOX所致APP/PS1 N2a细胞ROS的产生。结论：E2α可以减少细胞外Ap的产生,其机制可能与其抗氧化损伤有关。更多还原

【Abstract】 Objective To investigate the effects of 17a-estradiol (E2a) on the release ofβ-amyloid protein (Aβ) with the transgenic cell line APP/PS 1 N2a, and discuss the potential mechanism of such effects.Methods APP/PS 1 N2a cell lines were pretreated with 10 nM E2a for 2 days, then changed the culture medium, and treated with different methods:(1) cell lines were treated with either E2a (25nM,50nM, 100nM,200nM) or vehicle, The protein level of Aβand BACE1 both intracellular and extracellular of APP/PS 1 N2a cells were detected by Western blot. (2) cell lines were treated with 100nM E2a,20 mM glucose and 5 mU glucose oxidase(GOX), followed by exposure to the glucose oxidase (G/Gox) to induce oxidative stress. After 24 hours, the change of the ROS contents in APP/PS 1 N2a cells was observed for DHE.Results Pretreatment of APP/PS 1 N2a cells with different concentration (25nM,50nM, 100nM,200nM) E2a significantly reduced extracellular levels of AP(P<0.05). There was no change both in the intracellular levels of Aβand the levels of BACE1(P>0.05). E2a can reduce the ROS produced by APP/PS 1 N2a cells after G/Gox treatment.Conclusion E2a can reduce the production of extracellular Aβ,its mechanism may be related to the oxidation damage.更多还原