姜黄素抑制Cu2＋诱导的转APP695基因SH-SY5Y细胞氧化损伤和凋亡作用

doi:10.7506/spkx1002-6630-201703027

摘要/Abstract

摘要： 目的：研究Cu2＋诱导的转β-淀粉样前体蛋白（amyloid-β precursor protein，APP）基因SH-SY5Y（SHSY5Y-APP695）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 Cu2＋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（pSer40-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，Cu2＋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，pSer40-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，pSer40-Nrf2和HO-1表达量减少。结论：姜黄素在一定程度上减弱了Cu2＋诱导的氧化损伤和细胞凋亡作用。

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

Abstract: 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.

Key words: Alzheimer’s disease, Cu2+, oxidative damage, cell apoptosis, curcumin, Nrf2-antioxidant response element pathway

中图分类号:

郑博闻，姜招峰，黄汉昌. 姜黄素抑制Cu2＋诱导的转APP695基因SH-SY5Y细胞氧化损伤和凋亡作用[J]. 食品科学, doi: 10.7506/spkx1002-6630-201703027.

ZHENG Bowen, JIANG Zhaofeng, HUANG Hanchang. Inhibitory Effects of Curcumin on Cu2+-Induced Oxidative Damage and Cell Apoptosis in Transgentic APP695 SH-SY5Y Cells[J]. FOOD SCIENCE, doi: 10.7506/spkx1002-6630-201703027.

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姜黄素抑制Cu2＋诱导的转APP695基因SH-SY5Y细胞氧化损伤和凋亡作用

Inhibitory Effects of Curcumin on Cu2+-Induced Oxidative Damage and Cell Apoptosis in Transgentic APP695 SH-SY5Y Cells

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