【摘要】 目的:研究高浓度胰岛素对树突状细胞(Dendritic Cell,DC)分化、成熟及免疫功能的影响。方法:采用连续贴壁法分离正常人外周血单核细胞,在含重组人粒-巨噬细胞集落刺激因子(GM-CSF,100ng/ml)和重组人白细胞介素-4(IL-4,100ng/ml)的完全培养基中培养。5天后收集细胞,重新铺板后继续在胰岛素浓度分别为0nmol/L、10nmol/L及100nmol/L的培养基中培养48小时,收集细胞和上清液,采用流式细胞术检测细胞表面CD40、CD80和CD83的表达;用ELISA法检测上清液中细胞因子IL-12、IL-10、TNF-α的浓度;用倒置显微镜动态观察DC形态变化。结果:胰岛素浓度为10nmol/L、100nmol/L组的DC表面标成CD40、CD80和CD83阳性表达率较含胰岛素0nmol/L的对照组升高,培养上清液中细胞因子IL-12、TNF-α的浓度也较对照组升高,而细胞因子IL-10的浓度则较对照组降低。结论:高浓度胰岛素促进了树突状细胞表型CD40、CD80和CD83的表达;促进了DC对细胞因子IL-12和TNF-α的分泌;对IL-10的分泌则起抑制作用。高浓度胰岛素通过促进树突状细胞免疫功能的成熟,可能是其参与动脉粥样硬化免疫炎症反应的发生、发展的机制之一。更多还原

【Abstract】 Objective To explore the effect of hyperinsulin on the maturation of monocyte-derived dendritic cells (DC).Methods Human monocytes were purifed using successive adherence method,after 5 days’ culture in RPMI1640 medium containing granulocyte-macrophage colony stimulating factor (GM-CSF,100ng/ml) and interleukin-4(IL-4,100ng/ml).Immature DC were then incubated with various concentrations (0,10,100nmol/L) insulin for 48 hours.Immunophenotypic expression of CD40,CD80 and CD83 was detected using flow cytometry.Mature DC secretions of IL-12,IL-10 and TNF-α were measured by ELISA.The utralmicro structure of dendritic cells was observed by microscope.Results Hyperinsulin promoted CD40,CD80 and CD83 expression and enhanced to excrete cytokine secretions of IL-12 and TNF-α significantly and decreased the excrete cytokines IL-10 secretion in healthy human.Conclusions Hyperinsulin contributed to atherosclerosis via stimulating immune maturation of mature DC,which may be one of its mechanisms of AS.更多还原