胍丁胺对脂多糖诱导RAW264.7细胞氧化应激损伤的保护作用  

Protective effects of agmatine on oxidative stress induced by LPS in RAW264.7 cells

作　　者：侯凤艳[1] 罗莉[2] 朱俊宇[2] 康文渊[2] 范霞[2] 柴鉴深[2] 卢中秋[1] 梁华平[1,2] 

机构地区：[1]温州医科大学附属第一医院急诊科,浙江温州325035 [2]第三军医大学大坪医院野战外科研究所第一研究室,创伤、烧伤与复合伤国家重点实验室,重庆400042

出　　处：《第三军医大学学报》2016年第9期949-953,共5页Journal of Third Military Medical University

基　　金：国家重点基础研究发展计划(973计划,2012CB518102)~~

摘　　要：目的探讨胍丁胺(agmatine,AGM)对脂多糖(lipopolysaccharides,LPS)诱导小鼠巨噬细胞系RAW264.7氧化应激的抑制作用及其机制。方法 RAW264.7细胞经LPS(10μg/m L)刺激发生氧化应激,同时AGM(1 mmol/L)对其进行干预。分为对照组、LPS组、LPS+AGM组、AGM组。药物作用24 h,以2,7-二氢二氯荧光素二乙酸酯(2',7'-dichlorofluoresce in diacetate,DCFH-DA)为荧光探针检测细胞内活性氧(reactive oxygen species,ROS)水平;Griess法检测细胞内一氧化氮(NO)水平;Western blot检测细胞内的iNOS蛋白及细胞质细胞核内Nrf2蛋白表达;RT-PCR检测Nrf2下游抗氧化酶血红素氧合酶(heme oxygenase-1,HO-1)mRNA的表达。结果与对照组相比,LPS可显著增加RAW264.7细胞中ROS、NO以及iNOS蛋白水平,而AGM干预后上述指标含量均明显降低(P<0.05)。AGM干预组中细胞核的Nrf2表达及其下游分子HO-1 mRNA表达水平较LPS单独刺激组显著升高,表明Nrf2信号通路在AGM的作用下被活化。结论 AGM可通过活化转录因子Nrf2促进抗氧化酶HO-1的表达,进而减轻细胞内的氧化应激损伤。Objective To determine the inhibitory effect of agmatine（ AGM） on lipopolysaccharides（ LPS）-induced oxidative stress in mouse macrophage RAW264. 7 cells and its underlying mechanism.Methods RAW264. 7 cells were stimulated by LPS（ 10 μg / m L） to induce oxidative stress,and treated with1 mmol / L AGM. Thus the cells were divided into control,LPA,LPS ＋ AGM,and AGM groups. In 24 h after the treatment,reactive oxygen species（ ROS） level of these cells was determined with 2,7-dichlorofluoresce in diacetate（ DCFH-DA） as fluorescence probe,intracellular NO level by Greiss method,intracellular i NOS protein and cytoplasmic / nuclear Nrf2 protein expression by Western blotting,and mRNA expression of heme oxygenase-1（ HO-1） by RT-PCR. Results Compared with the control group,LPS significantly induced the levels of ROS,NO and iNOS protein in RAW264. 7 cells,and AGM significantly reversed such effect（ P〈0. 05）. Nrf2 pathway and its target HO-1 were significantly up-regulated compared to LPS-stimulated cells,suggesting that AGM activated the Nrf2 signaling pathway. Conclusion AGM reduces oxidative damage of the RAW264. 7 cells by promoting HO-1 antioxidase expression through activation of the Nrf2 pathway.

关 键 词：胍丁胺 RAW264.7细胞 氧化应激 NRF2 

分 类 号：R392.12[医药卫生—免疫学] R966[医药卫生—基础医学]