姜黄素抑制Cu^(2+)诱导的转APP_(695)基因SH-SY5Y细胞氧化损伤和凋亡作用  

Inhibitory Effects of Curcumin on Cu^(2+)-Induced Oxidative Damage and Cell Apoptosis in Transgentic APP_(695) SH-SY5Y Cells

作　　者：郑博闻[1] 姜招峰[1] 黄汉昌[1] ZHENG Bowen;JIANG Zhaofeng;HUANG Hanchang(Beijing Key Laboratory of Bioactive Substances and Functional Foods, Beijing Union University, Beijing 100191, China)

机构地区：[1]北京联合大学生物活性物质与功能食品北京市重点实验室,北京100191

出　　处：《食品科学》2017年第3期164-169,共6页Food Science

基　　金：国家自然科学基金面上项目(31471587);北京市属高等学校高层次人才引进与培养计划项目(CIT&TCD201504034)

摘　　要：目的:研究Cu^(2+)诱导的转β-淀粉样前体蛋白(amyloid-β precursor protein,APP)基因SH-SY5Y(SHSY5Y-APP_(695))细胞氧化损伤、凋亡及姜黄素的抑制作用。方法:在姜黄素预保护和无姜黄素预保护条件下,50 μmol/L Cu^(2+)处理细胞24 h,测定细胞存活率、胞外乳酸脱氢酶(lactate dehydrogenase,LDH)水平、胞内活性氧(reactive oxygen,ROS)水平、线粒体膜电位、半胱氨酸天冬氨酸蛋白酶(caspase)-3、caspase-8和caspase-9活力,蛋白免疫印迹法测定核转录NF-E2相关因子2(NF-E2-related factor 2,Nrf2)Ser40位点磷酸化(p Ser40-Nrf2)水平以及血红素加氧酶(heme oxygenase,HO)-1蛋白表达水平。结果:与空白对照相比,Cu^(2+)损伤组细胞存活率降低,胞内ROS水平和胞外LDH活性升高,线粒体膜电位下降,caspase-9和caspase-3活性明显升高,p Ser40-Nrf2和HO-1含量增加,而姜黄素保护组细胞存活率升高,胞内ROS水平和LDH释放水平降低,线粒体膜电位恢复升高,caspase-9和caspase-3活性明显降低,p Ser40-Nrf2和HO-1表达量减少。结论:姜黄素在一定程度上减弱了Cu^(2+)诱导的氧化损伤和细胞凋亡作用。Objective: This study was focused on the protective effect of curcumin on cellular oxidative damage and cell apoptosis induced by Cu2+ in human neuroblastoma SH-SY5Y cells transfected by human amyloid-β precursor protein (APP) (SH-SY5Y-APP695). Methods: SH-SY5Y-APP695 cells were treated with 50 μmol/L Cu2+ at 37 ℃ for 24 h with or without curcumin pre-protection. Cell viability was detected by cell counting kit-8 (CCK-8). Extracellular lactate dehydrogenase (LDH), intracellular reactive oxygen species (ROS) and mitochondrial membrane potential were determined by commercial assay kits. The enzymatic activities of caspase-3, caspase-8 and caspase-9 were assessed. Phosphorylation levels of NF-E2-related factor 2 (Nrf2) at Ser-40 (pSer40-Nrf2) and HO-1 were detected by Western blot analysis. Results: Compared with the control group, Cu2+ administration led to decreased cell viability and mitochondrial membrane potential, increased levels of LDH, intracellular ROS and caspase-3, caspase-8 and caspase-9 activities, and elevated levels of pSer40-Nrf2 and HO-1.Conversely, curcumin increased cell viability and mitochondrial membrane potential, decreased the levels of LDH and intracellular ROS, significantly mitigated caspase-9 and caspase-3 activities, and reduced the expression levels pSer40-Nrf2 and HO-1. Conclusion: Curcumin can attenuate Cu2+-induced cellular oxidative damage and cell apoptosis.

关 键 词：阿尔茨海默病 CU2+ 氧化损伤 细胞凋亡 姜黄素 Nrf2/ARE信号通路 

分 类 号：Q26[生物学—细胞生物学]