柿叶提取物对HEK293-APPswe转基因细胞模型的抗氧化作用及对Nrf2/HO-1途径的影响  

Antioxidant effect of persimmon leaf extract to HEK293-APPswe transgenic cells and effect to Nrf2/HO-1 pathway

作　　者：吴小凡[1] 马斌[2] 侯训尧[1] 洪艳[1] 申超[1] 刘雪平[1] WU Xiao-Fan MA Bin HOU Xun- Yao HONG Yan SHEN Chao LIU Xue-Ping(Department of Senile Neurology, Shandong Provincial Hospital Affiliated to Shandong University, Ji＇nan 250021, China)

机构地区：[1]山东大学附属省立医院老年神经科,济南250021 [2]山东大学药学院,济南250012

出　　处：《中国免疫学杂志》2017年第6期854-858,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金(81371225);山东省科技发展计划(2014GSF118056)项目资助

摘　　要：目的:观察柿叶提取物(PLE)对阿尔茨海默病细胞模型HEK293-APPswe(20E2)的抗氧化应激的影响,并从核因子E2相关因子2(Nrf2)/血红素加氧酶1(HO-1)信号通路研究其作用机制。方法:Western blot检测APP蛋白表达水平,ELISA检测各组细胞上清Aβ_(1-40)的量,确定细胞模型是否建立成功。用CCK-8检测不同浓度的柿叶提取物对细胞活性的影响,选定干预最佳浓度。设分组:SH-SY5Y为空白对照组(NC组),20E2为模型组(20E2组),用柿叶提取物干预为加药组(20E2+PLE组)。DCFH-DA荧光探针检测各组细胞内ROS的变化,ELISA检测各组细胞上清Aβ_(1-42)的量,Western blot检测胞核、胞浆Nrf2和全细胞HO-1蛋白的表达水平变化。结果:与模型组相比,加药组ROS表达减少,细胞外Aβ_(1-42)浓度降低,细胞核内Nrf2表达增多,HO-1蛋白含量增加。结论:柿叶提取物能有效降低模型组氧化应激的水平,可能是通过降低Aβ_(1-42)的聚集,激活Nrf2/HO-1信号途径,促进Nrf2合成和核转位,从而促进下游抗氧化蛋白HO-1的表达来减弱氧化应激的损伤。氧化应激的影响,并从测APP蛋白表达水平,提取物对细胞活性的影提取物干预为加药组量,Western blot检测胞外Aβ_(1-42)浓度降低,细胞是通过降低Aβ_(1-42)的聚氧化应激的损伤。Objective：To investigate the effect of persimmon leaf extract （PLE） on HEK293-APPswe transgenic cells （20E2）. Methods：To determine whether the 20E2 cells model was successfully established,the level of Aβ1-42 in SH-SYSY was detected and 20E2 cells （HEK293 cells stably expressing Swedish mutant APP） cultured in vivo by ELISA kit, and the expression of APP protein level was detected by Western blot. Cell viability was assayed by CCK-8 method and then selected the best concentration. Set groups： SH-SY5Y as normal control group （NC group） ,20E2 as model group （20E2 group） ,treating with PLE as treating group （20E2＋PLE group）. Reactive oxygen species （ROS） levels of each group were detected with DCFH-DA fluorescent probe. The extracellular level of Aβ1-42 were detected by ELISA kit, Cytoplasmic Nrf2, Nuclear Nrf2, Whole-cell HO-1 were detected by Western blot. Results：Compared with model group, the expressions of ROS, Aβ1-42 were down-regulated and the Nuclear Nrf2 and Whole-cell HO-1 were up-regulated in 20E2 ＋PLE group. Conclusion ：PLE can reduce the level of oxidative stress of model group effectively, it possibly reduce the aggregation of Aβ1-42 and prevent oxidizing via activating Nrf2/HO-1 pathway.

关 键 词：柿叶 氧化应激 神经保护 Nrf2/HO-1 

分 类 号：R741.05[医药卫生—神经病学与精神病学]