去甲肾上腺素诱导人巨噬细胞白细胞介素6表达及其机制

发布时间：2018-03-21 15:59

本文选题：去甲肾上腺素　切入点：巨噬细胞　出处：《中国药理学与毒理学杂志》2017年03期 　论文类型：期刊论文

【摘要】：目的观察去甲肾上腺素(NE)预处理人单核/巨噬细胞U937细胞白细胞介素6(IL-6)的表达,探讨其致动脉粥样硬化可能的作用机制。方法 NE 0.01~10μmol·L~(-1)与U937细胞共培养24 h后,RT-PCR法检测细胞IL-6 mRNA水平;共培养6,9,12,24和48 h后,ELISA法检测细胞培养液上清中IL-6蛋白水平;共培养24 h后,荧光探针法检测细胞内活性氧(ROS)的生成。预先加入ROS抑制剂N-乙酰半胱氨酸(NAC)、NADPH氧化酶抑制剂二亚苯基碘(DPI)或线粒体复合物Ⅱ抑制剂噻吩甲酰三氟丙酮(TIFA)孵育1 h后,再加入NE 0.01~10μmol·L~(-1)继续培养24 h,ELISA法检测细胞培养液上清中IL-6蛋白水平。结果巨噬细胞IL-6 mRNA表达和蛋白水平随NE 0.01~10μmol·L~(-1)浓度增加和孵育时间延长而升高,NE 1.0μmol·L~(-1)刺激24 h,IL-6 mRNA表达和蛋白水平分别为细胞对照组的2.62和4.47倍(P0.01);NE 0.01,0.1,1.0和10.0μmol·L~(-1)组ROS的生成分别为对照组的1.87,2.56,2.91和5.36倍(P0.01)。NAC 10 mmol·L~(-1)和DPI 10μmol·L~(-1)均一定程度地抑制IL-6蛋白的表达(P0.01),而TIFA无影响。结论 NE可诱导人巨噬细胞IL-6的表达,并可能通过NADPH氧化酶介导的ROS通路而促进炎症反应,促进动脉粥样硬化的发生和发展。
[Abstract]:Objective to observe the expression of interleukin 6 (IL 6) in U937 cells pretreated with noradrenaline (NE). Methods the levels of IL-6 mRNA were detected by RT-PCR after co-cultured with U937 cells for 24 h, and the levels of IL-6 protein in supernatant of cultured cells were detected by Elisa after co-culture for 24 h and 48 h after co-culture, and after co-culture for 24 h, the levels of IL-6 protein in the supernatant of U937 cells were detected by Elisa. Fluorescence probe assay was used to detect the formation of reactive oxygen species (Ros) in cells. After 1 h incubation with ROS inhibitor N-acetylcysteine, NADPH oxidase inhibitor, diphenyl iodide (DPI), or mitochondrial complex inhibitor, thiophenyl trifluoroacetone, TIFA (1 h). The expression and protein level of IL-6 mRNA in the supernatant of macrophages were determined by Elisa. Results the expression of IL-6 mRNA and the protein level of macrophages increased with the increase of NE 0.01 10 渭 mol 路L ~ (-1) concentration and the incubation time increased. NE 1.0 渭 mol 路L ~ (-1)) stimulated the expression of IL-6 mRNA and the expression of IL-6 mRNA at 24 h. The protein levels were 2.62 and 4.47 times of P0.01ANE0.01P0.01P0.01 and 10.0 渭 mol 路L ~ (-1) in the control group, respectively. The ROS production in the control group was 1.87 渭 mol 路L ~ (-1) and 5.36 times that of the control group respectively (P 0.01U. NAC 10 mmol 路L -1) and DPI (10 渭 mol 路L ~ (-1)) could inhibit the expression of IL-6 protein to some extent, but TIFA had no effect. Conclusion NE can induce the expression of IL-6 protein to a certain extent, but TIFA has no effect. Leading to the expression of IL-6 in human macrophages, ROS pathway mediated by NADPH oxidase may promote inflammatory response and promote the occurrence and development of atherosclerosis.
【作者单位】：南京中医药大学连云港附属医院连云港市中医院;济宁医学院附属医院;
【基金】：江苏省药学会-奥赛康医院药学基金(201517) 连云港市科技发展计划(ZD1508) 江苏省青年医学人才(QNRC2016507)~~
【分类号】：R96

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