摘要
目的 探讨17α雌二醇(ESUB>2/SUB>α)对稳定表达人β-淀粉样前体蛋白(APP)/早老因子-1(PS1)即APP /PS1基因的神经母细胞瘤细胞株(Neuro-2a)(APP/PS1 NSUB>2/SUB>a)β-淀粉样蛋白(Aβ)产生的影响及可能机制。方法 将APP/PS1 NSUB>2/SUB>a细胞用10×10SUP>-9/SUP> mol/L E2α预处理2d,更换培养液后,分别进行如下处理:1.加入不同浓度ESUB>2/SUB> a(25nmol/L、50nmol/L、100nmol/L、200nmol/L)和溶媒(对照组),用免疫印迹法(Western blotting)检测APP/PS1 NSUB>2/SUB> a细胞内和细胞外的Aβ蛋白水平及细胞BACE1蛋白水平;2.加入 100nmol/L ESUB>2/SUB>α、20nmol/L葡萄糖和5mIU葡萄糖氧化酶(GOX)。24h后,进行二氢乙啶(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(25nmol/L、50nmol/L、100nmol/L、200nmol/L)处理后,细胞外Aβ蛋白水
Abstract
Objective To investigate the effects of 17α-estradiol (ESUB>2/SUB>α) on the release of β-amyloid protein (Aβ) with the transgenic cells β-amyloid precursor protein(APP)/ presenilin-1(PS1)Neuro-2a (NSUB>2/SUB>a), and explore the potential mechanism of such effect. Methods APP/PS1 NSUB>2/SUB>a cells were pretreated with 10×10SUP>-9/SUP> mol/L Eα for 2 days, then changed the culture medium, and treated with different methods: 1. Cells were treated with either E2α(25 nmol/L, 50 nmol/L, 100 nmol/L, 200 nmol/L)or vehicle, the protein level of Aβ and β-secretase-1(BACE1) both intracellular and extracellular of APP/PS1 N2a cells were detected by Western blotting. 2. Cells were treated with 100 nmol/L E2α, 20 nmol/L glucose and 5 IU glucose oxidase(GOX), followed by exposure to the glucose oxidase (G/Gox) to induce oxidative stress. After 24 hours, the change of the Reactive oxygen species(ROS)contents in APP/PS1 NSUB>2/SUB>a cells was observed for dihydroethidium(DHE). Results The extracellular level of Aβin APP/PS 1 NSUB>2/SUB>a cells treated with various concentration (25 nmol/L,50 nmol/L,100 nmol/L,200 nmol/L/L)ESUB>2/SUB>α were significantly decreased(EM>P/EM> <0.05). There was no change in the intracellular levels of Aβ and the levels of BACE1(EM>P/EM> >0.05). ESUB>2/SUB>α reduced the ROS produced by APP/PS1 NSUB>2/SUB>a cells after G/Gox treatment. Conclusion ESUB>2/SUB>α may reduce the production of extracellular Aβ and its mechanism may be related
关键词
17α雌二醇 /
神经母细胞瘤细胞株 /
β-淀粉样前体蛋白/早老因子-1 /
β-淀粉样蛋白 /
免疫印迹法 /
二氢乙啶染色
Key words
17α-estradiol /
Neuro-2a /
APP/PS1 /
β-amyloid /
Western blotting /
Dihydroethidium staining
李建明;曾杰;罗学港;雷德亮.
17α雌二醇对转基因细胞株(APP/PS1N2a)β-淀粉样蛋白生成的影响 [J]. 解剖学报. 2012, 43(6): 772-777 https://doi.org/10.3969/j.issn.0529-1356.2012.06.010
Effects and possible mechanisim of 17α-estradiol on the release ofβ-amyloid protein with the transgenic cell strain APP/PS1 NSUB>2/SUB>a [J]. Acta Anatomica Sinica. 2012, 43(6): 772-777 https://doi.org/10.3969/j.issn.0529-1356.2012.06.010
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