脂氧素类似物BML-111对炎症状态人单核细胞株U937氧化应激标志物表达的影响  

Effect of BML-111 on expression of markers of inflammation-induced oxidant stress in human monocytes U937

作　　者：郝建[1] 郝华[2] 徐芬[2] 徐静 张建[2] 李里香[2] 曾磊[2] HAO Jian;HAO Hua;XU Fen;XU Jing;ZHANG Jian;LI Lixiang;ZENG Lei(The Third People＇s Hospital of Bengbu,Bengbu 233000,Chin)

机构地区：[1]蚌埠市第三人民医院,安徽蚌埠233000 [2]南昌大学第二附属医院

出　　处：《山东医药》2018年第23期22-25,共4页Shandong Medical Journal

基　　金：国家自然科学基金项目(81301741);江西省自然科学基金项目(20151BAB205038;20171BAB205054);江西省科技支撑计划项目(20161BBG70201);南昌大学第二附属医院青年基金项目(2016YNQN12017)

摘　　要：目的观察脂氧素类似物BML-111预处理对炎症状态人单核细胞株U937氧化应激标志物表达的影响。方法培养U937细胞并分为对照组、模型组、实验组。对照组不加入药物;模型组加入1μg/L的脂多糖(LPS)制作炎症模型;实验组先加入200μg/L的BML-111预处理30 min后再加入1μg/L的LPS。采用细胞免疫荧光法检测各组U937细胞中的核因子E2相关因子2(Nrf2);采用RT-PCR法检测各组U937细胞中的Nrf2、醌氧化还原酶(NQO1)、血红素加氧酶1(HO-1)、TNF-αmRNA。结果对照组细胞中Nrf2蛋白主要定位于胞核与胞质;模型组细胞中Nrf2蛋白主要定位于胞核,即从胞质转位至核内;实验组Nrf2蛋白表达定位与对照组相近。模型组Nrf2蛋白相对表达量高于对照组、实验组Nrf2蛋白相对表达量低于模型组(P均<0.05),模型组中Nrf2、NQO1、HO-1、TNF-αmRNA相对表达量高于对照组、实验组细胞中Nrf2、NQO1、HO-1、TNF-αmRNA相对表达量低于模型组(P均<0.05)。结论 BML-111预处理后,炎症状态的U937细胞中Nrf2蛋白核转位受抑制,Nrf2蛋白及其下游基因表达下调,TNF-α基因表达下调;调节Nrf2蛋白及其下游基因表达、下调TNF-α表达可能是BML-111抗氧化应激作用机制的一部分。Objective To investigate the effect of the analog of lipoxin BML-111 pretreatment on the expression of markers of inflammation-induced oxidant stress in the human monocytes U937. Methods U937 cells were randomly divided into three groups： the control group,lipopolysaccharide（ LPS） group（ model group）,and LPS ＋ BML-111 group（ experimental group）. We did not add any drugs to the control group; in the model group,1 μg/L LPS was added to make an inflammation model; the cells in the experimental group were first treated with 200 μg/L BML-111 for 30 min and then were treated with 1 μg/L LPS. The nuclear factor-E2-related factor 2（ Nrf2） expression was detected by immunofluorescence,and RT-PCR was used to detect the Nrf2,NADH quinone oxidoreductase 1（ NQO1）,heme oxygenase-1（ HO-1）and TNF-α mRNA in the U937 cells. Results In the control group,Nrf2 protein was mainly located in the nucleus and cytoplasm; in the model group,Nrf2 protein was mainly located in the nucleus,from the cytoplasm to the nucleus; the Nrf2 protein localization in the experimental group was similar to that in the control group. The relative expression of Nrf2 protein in the model group was higher than that in the control group,and the relative expression of Nrf2 protein in the experimental group was lower than that in the model group（ both P〈0. 05）. The relative expression levels of Nrf2,NQO1,HO-1 and TNF-α mRNA in the model group were higher than those in the control group; the relative expression levels of Nrf2,NQO1,HO-1 and TNF-α mRNA in the experimental group were lower than those in the model group（ all P〈0. 05）. Conclusion After pretreatment with BML-111,nuclear translocation of Nrf2 protein is inhibited in the inflammatory U937 cells,the Nrf2 protein is down-regulated,and its downstream gene expression is up-regulated; regulating the Nrf2 protein and its downstream gene expression and down-regulating the TNF-α expression may be part of the mechanisms in anti-oxidative stress of BML-111.

关 键 词：单核细胞 脂氧素类似物 炎症 氧化应激 核因子E2相关因子2 醌氧化还原酶 血红素加氧酶1 肿瘤坏死因子α 

分 类 号：R363.2[医药卫生—病理学] R631[医药卫生—基础医学]