氢气对氧化损伤结肠腺癌细胞的保护作用及机制  

Protective effect of hydrogen saline in oxidative stress insulted enterocytes

作　　者：穆怀博[1,2] 盛庆丰[1,2] 吴伟[1,2] 吕志葆[1,2] 

机构地区：[1]上海交通大学附属儿童医院,上海220000 [2]上海市儿童医院

出　　处：《山东医药》2015年第6期4-7,共4页Shandong Medical Journal

基　　金：上海市科委引导类重点课题(124119a6500)

摘　　要：目的间接探讨氢气对氧化损伤肠上皮细胞的保护作用及其机制。方法取结肠腺癌Caco-2细胞与IEC-6细胞株,分别分为对照组、模型组及观察组。对照组采用常规RPMI1640培养基、模型组采用含抗霉素A的RPMI1640培养基、观察组采用含氢气及抗霉素A的RPMI1640培养基培养。检测各组以下指标:1氧自由基(O-2·、H2O2和·OH)水平;2线粒体膜电位;3氧化代谢产物丙二醛(MDA)和8-羟基鸟嘌呤(8-OH-G);4细胞凋亡率和乳酸脱氢酶(LDH)活性;5促炎症因子(IL-6、TNF-α、IL-1β)、抗炎症因子(IL-10)、转录因子核因子-E2相关因子2(Nrf2)及其下游靶基因(HO-1、SOD、Cat、GPx)表达;6磷酸化Nrf2蛋白表达。结果与模型组比较,观察组细胞内氧自由基水平降低,线粒体膜电位升高,细胞凋亡率及LDH活性降低,促炎症因子、抗炎症因子、转录因子Nrf2及其下游靶基因表达增强,磷酸化Nrf2蛋白表达增强。结论氢气对氧化损伤的结肠腺癌细胞有保护作用;其机制可能与调节Nrf2/ARE通路有关。根据本研究结果推测氢气对氧化损伤肠上皮细胞有保护作用。Objective To explore the effect of hydrogen saline in oxidative stress insulted enterocytes .Methods Caco-2 and IEC-6 cells were divided as 3 groups：control, model and hydrogen treated groups .Cells in control group was treated with normal RPMI1640 medium.Model group was treated with RPMI1640 medium containing antimycin A .Cells in hydrogen treated groups was treated with RPMI 1640 medium containing antimycin A and H 2.①Levels of O2^-· 、H2 O2 adn · OH;②Mitochondrial membrane potential;③MDA and 8-OH-G;④Cell apoptoiss and LDH activity;⑤Proinflammatory factors（IL-6、TNF-α、IL-1β）、anti-inflammatory factors（IL-10）,expressions of Nrf2 and the downstream target genes （HO-1、SOD、Cat、GPx）;⑥Expression of phosphorylation-Nrf2 were detected for further study .Results We found that H2 ef-fectively reduce the reactive oxygen specis , cell apoptosis and activity of LDH in culture medium and improved the mito-chondrial membrane potential .H2 activated the phosphorylation of Nrf 2.H2 significantly reduced the release of inflamma-tory cytokines and promoted the downstream genes expressions of Nrf 2.Conclusions Our work demonstrated that H2 showed favorable ability in necrotizing enterocolitis treatment .The mechanism was be closely related with Nrf 2-ARE signa-ling.

关 键 词：坏死性小肠结肠炎 结肠腺癌 氢气 Nrf2-ARE信号通路 氧化应激反应 

分 类 号：R34[医药卫生—基础医学]