胍丁胺对脂多糖诱导RAW264.7细胞氧化应激损伤的保护作用

发布时间：2018-08-26 07:52

【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(P0.05)。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。
[Abstract]:Objective to investigate the inhibitory effect of agmatine (agmatine,AGM) on oxidative stress induced by lipopolysaccharide (lipopolysaccharides,LPS) in mouse macrophage cell line RAW264.7 and its mechanism. Methods oxidative stress was induced by LPS (10 渭 g / mL) in RAW264.7 cells, and AGM (1 mmol/L) interfered with it. The rats were divided into two groups: control group, lipopolysaccharide group, AGM group. For 24 h, the level of intracellular reactive oxygen species (reactive oxygen species,ROS) was detected with 2H7-dichlorofluoresce in diacetate,DCFH-DA as fluorescence probe. The level of nitric oxide (NO) in cells was detected by Griess method. Western blot was used to detect the level of iNOS protein and fine cytoplasm in cells. The expression of Nrf2 protein in nucleus was detected by RT-PCR. The expression of heme oxygenase (heme oxygenase-1,HO-1) mRNA in the downstream of Nrf2 was detected by reverse transcriptase polymerase chain reaction (RT-PCR). Results compared with the control group, lipopolysaccharide could significantly increase the levels of ROS,NO and iNOS protein in RAW264.7 cells. The expression of nuclear Nrf2 and its downstream HO-1 mRNA expression in AGM intervention group were significantly higher than those in LPS alone group, indicating that Nrf2 signaling pathway was activated by AGM. Conclusion AGM can promote the expression of antioxidant enzyme HO-1 by activating transcription factor Nrf2, and then alleviate the oxidative stress injury in cells.
【作者单位】：温州医科大学附属第一医院急诊科;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;
【基金】：国家重点基础研究发展计划(973计划,2012CB518102)~~
【分类号】：R965

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