CAJ | 학술논문

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（O2－·、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。
목적：간접탐토경기대양화손상장상피세포적보호작용급기궤제。방법취결장선암Caco-2세포여IEC-6세포주，분별분위대조조、모형조급관찰조。대조조채용상규RPMI1640배양기、모형조채용함항매소A적RPMI1640배양기、관찰조채용함경기급항매소A적RPMI1640배양기배양。검측각조이하지표：①양자유기（O2－·、H2O2화· OH）수평；②선립체막전위；③양화대사산물병이철（MDA）화8-간기조표령（8-OH-G）；④세포조망솔화유산탈경매（ LDH）활성；⑤촉염증인자（ IL-6、TNF-α、IL-1β）、항염증인자（ IL-10）、전록인자핵인자-E2상관인자2（Nrf2）급기하유파기인（HO-1、SOD、Cat、GPx）표체；⑥린산화Nrf2단백표체。결과여모형조비교，관찰조세포내양자유기수평강저，선립체막전위승고，세포조망솔급LDH활성강저，촉염증인자、항염증인자、전록인자Nrf2급기하유파기인표체증강，린산화Nrf2단백표체증강。결론경기대양화손상적결장선암세포유보호작용；기궤제가능여조절Nrf2／ARE통로유관。근거본연구결과추측경기대양화손상장상피세포유보호작용。
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.