中文摘要：

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 ＋ H2S 100 μmol/L组及ox-LDL ＋ H2S 500 μmol/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 100 μmol/L组及ox-LDL ＋ H2S 500 μmol/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 100 μmol/L组及ox-LDL ＋ H2S 500 μmol/L组细胞胞浆IκBα表达显著增多，NF-κB p65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 100 μmol/L组及ox-LDL ＋ H2S 500 μmol/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?

英文摘要：

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