H_2S对氧化型低密度脂蛋白诱导人单核巨噬细胞核转录因子-κB的影响及机制  被引量:1

Impact of H_2S on oxidized-low density lipoprotein-stimulated nuclear factor-κB in human monocytes/macrophage and its mechanisms

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作  者:赵曼曼[1] 张巧丽[1] 闫辉[1] 杜军保[1] 耿彬[2] 唐朝枢[2,3] 金红芳[1] 

机构地区:[1]北京大学第一医院儿科,北京100034 [2]北京大学基础医学院生理与病理生理学系,北京100191 [3]教育部分子心血管学重点实验室,北京100191

出  处:《北京大学学报(医学版)》2013年第2期192-196,共5页Journal of Peking University:Health Sciences

基  金:国家自然科学基金(31130030和81070212)资助~~

摘  要:目的:研究硫化氢(hydrogen sulfide,H2S)对氧化型低密度脂蛋白(oxidized low-density lipoprotein,ox-LDL)诱导人单核巨噬细胞NF-κB通路的调节作用及其机制。方法:人单核细胞系THP-1细胞采用12-豆蔻酸-13-乙酸佛波醇(phorbol myristate acetate,PMA)诱导分化为巨噬细胞后,分为4组:对照组、ox-LDL组、ox-LDL+H2S100μmol/L组及ox-LDL+H2S 500μmol/L组。用Western blot检测细胞IκBα蛋白表达及NF-κB磷酸化水平,激光共聚焦法检测细胞胞浆IκBα及NF-κB的核转位改变,免疫共沉淀方法检测细胞核中NF-κB p65与IκBα结合情况。结果:Western blot结果显示,与对照组相比,ox-LDL组人单核巨噬细胞中NF-κB p65磷酸化水平明显升高(0.855±0.116 vs.0.502±0.218,P=0.046),IκBα表达明显减少(0.612±0.216 vs.0.997±0.167,P=0.029);与ox-LDL组相比,ox-LDL+H2S 100μmol/L组及ox-LDL+H2S 500μmol/L组细胞中NF-κB p65磷酸化水平显著降低(0.424±0.225 vs.0.855±0.116,P=0.020;0.378±0.071 vs.0.855±0.116,P=0.011),IκBα表达显著增多(1.037±0.111 vs.0.612±0.216,P=0.015;1.046±0.084 vs.0.612±0.216,P=0.013)。激光共聚焦结果显示:与对照组相比,ox-LDL组THP-1源性巨噬细胞胞浆中IκBα表达明显降低,NF-κB p65核转位明显增加;与ox-LDL组相比,ox-LDL+H2S 100μmol/L组及ox-LDL+H2S 500μmol/L组细胞胞浆IκBα表达显著增多,NF-κBp65核转位明显减少。免疫共沉淀结果显示,对照组人单核巨噬细胞胞核内未检测到NF-κB p65与IκBα的结合,ox-LDL组细胞核内NF-κB p65与IκBα结合较少,ox-LDL+H2S 100μmol/L组及ox-LDL+H2S 500μmol/L组细胞核内NF-κB与IκBα结合明显增多。结论:H2S可抑制ox-LDL诱导人单核巨噬细胞中NF-κB通路激活,其作用机制可能与抑制胞浆中IκBα降解,减少NF-κB p65磷酸化激活及核转位,同时可促进胞核中IκBα与NF-κB的结合,进而抑制NF-κB的活性有关。Objective: To investigate the role of hydrogen sulfide ( H2 S) on oxidized-low density lipo-protein (ox-LDL)-stimulated NF-κB pathway in human monocytes/macrophages and its mechanisms. Methods: THP-1 ceils were induced to differentiate into macrophages by incubation with phorbol myris-tate acetate (PMA) . The human monocytes/macrophages were divided into 4 groups, control group, ox-LDL group, ox-LDL + H2S 100 μmol/L group and ox-LDL + H2S 500 μmol/L group. The expression of IκBα and phosphorylation of NF-κB p65 in the cells were detected by Western blotting. The expression of IκBα and nuclear translocation of NF-κB p65 in the ceils were observed by laser confocal method. The interaction between NF-κB p65 and IKBot in the nuclear extracts was detected by coimmunoprecipitation method. Results: Compared with the control group, the phosphorylation of NF-κB p65 in the human mo-nocytes/macrophages of ox-LDL group was increased significantly (0. 855 ± 0. 116 vs. 0. 502±0.218, P =0.046), while the expression of IKBot in the cells of the ox-LDL group was decreased (0. 612 ±0. 216 vs. 0. 997±0. 167, P =0.029). Compared with the ox-LDL group, the phosphorylation of NF-κB p65 in the cells of the ox-LDL + H2S 100 μmol/L group and the ox-LDL + H2S 500 Ixmol/L group was decreased significantly (0.424 ±0. 225 vs. 0. 855 ±0. 116, P =0. 020; 0. 378± 0. 071 vs. 0. 855 ± 0. 116, P =0. 011, respectively), while the expressions of IκBα in the ceils of the ox-LDL + H2S 100 μmol/L group and the ox-LDL ± H2S 500 μmol/L group were increased (1.037 ±0. 111 vs. 0. 612 ± 0. 216, P =0. 015 ; 1. 046 ±0. 084 vs. 0. 612 ±0. 216, P =0. 013, respectively). The results from laser confocal method demonstrated that the IκBα expression in the cytoplasma of cells in the ox-LDL group was lower than that in the control group, and the nuclear translocation of NF-κB p65 in the cells of the ox-LDL group was higher than that in the control group. The IκBα expression in the cytopla

关 键 词:硫化氢 氧化低密度脂蛋白 NF-κB 单核细胞 巨噬细胞 

分 类 号:R331.3[医药卫生—人体生理学]

 

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