高浓度胰岛素对急性冠状动脉综合征患者外周单核细胞源树突状细胞分化成熟及免疫功能的影响

Effects of Hyperinsulin on Development and Maturation and Immune Function of Human Monocyte-derived Dendritic Cells in the Acute Coronary Syndrome Patients

目的研究高浓度胰岛素对急性冠状动脉综合征患者外周单核细胞源树突状细胞分化、成熟及免疫功能的影响。方法采用贴壁法分离急性冠状动脉综合征患者(ACS组)和正常人(正常组)外周血单核细胞,在含粒-巨噬细胞集落刺激因子(100μg/L)和白细胞介素4(100μg/L)的RPM I 1640完全培养基中培养。5天后收集细胞,作为未成熟树突状细胞,重新铺板后继续在胰岛素浓度分别为1、10 nmol/L和100 nmol/L的RPM I1640完全培养基中培养48 h后,收集细胞和上清液,此时细胞作为成熟树突状细胞,采用流式细胞术检测成熟树突状细胞表面CD40、CD80和CD83的表达;用酶联免疫吸附法测定检测上清液中细胞因子白细胞介素12、白细胞介素10和肿瘤坏死因子α的浓度;用倒置显微镜动态观察树突状细胞形态变化。结果树突状细胞表型CD40、CD80和CD83随着胰岛素浓度升高而升高(P<0.05),培养上清液中细胞因子白细胞介素12、肿瘤坏死因子α的浓度也随着胰岛素浓度升高而升高(P<0.05),而细胞因子白细胞介素10的浓度则随着胰岛素浓度升高而降低(P<0.05)。同等胰岛素浓度下,急性冠状动脉综合征患者较正常组的树突状细胞表面CD40、CD80和CD83的阳性表达率升高(P<0.05),培养上清液中细胞因子白细胞介素12、肿瘤坏死因子α的浓度升高(P<0.05),而细胞因子白细胞介素10的浓度则降低(P<0.05)。结论高浓度胰岛素促进了急性冠状动脉综合征患者的树突状细胞表面标志物CD40、CD80和CD83的表达;促进了树突状细胞对细胞因子白细胞介素12和肿瘤坏死因子α的分泌;对白细胞介素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成熟,参与动脉粥样硬化免疫炎症反应的发生和发展,是急性冠状动脉综合征发生的可能机制之一。

Aim To expore the effect of hyperinsulin on the maturation of monocyte-derived dendritic cells in the Acute Coronary Syndrome（ACS） patients. Methods Human monocytes of ACS were purifed using successive adherence method,recombinated human granulocyte-macrophage colony stimulating factor（GM-CSF,100 μg/L）and interleukin-4（IL-4,100 μg/L）after 5 days culture in RPMI 1640 medium containing.Immature DC（imDC） were collected,then added RPMI 1640 medium containing with insulin of various concentrations（1nmol/L,10nmol/L,100 nmol/L）for 48 hours,mature DC（mDC） were derived.Immunophenotypic expression of CD40,CD80 and CD83 were monitored by flow cytometry,and expression of IL-12,IL-10 and TNF-α were measured by ELISA,and the morphological features of dendritic cells were observed with invert optical microscope. Results Hyperinsulin promoted the expression of CD40,CD80 and CD83 and enhanced the expression of IL-12 and TNF-α significantly and refained the expression of IL-10 in the acute coronary syndrome patients. Conclusions Hyperinsulin contributed to the development of atherosclerosis via stimulating immune maturation of DC,which may be one of its mechanisms in the development of ACS.