糖基化终产物对人单核细胞源树突状细胞VCAM-1表达的影响

Effect of advanced glycosylation end products on the expression of VCAM-1 in human monocyte-derived dendritic cells

目的:探讨糖基化终产物(AGEs))对人单核细胞源树突状细胞(DCs)血管细胞黏附分子-1(VCAM-1)表达的影响。方法:用免疫磁珠分离人外周血CD14+单核细胞,经含重组人粒细胞-巨噬细胞集落刺激因子(rhGM-CSF)100μg/L和重组人白细胞介素-4(rhIL-4)20μg/L的RPMI1640培养,使其分化为DCs,加入糖基化人血清白蛋白(AGE-HSA)200μg/ml,采用RT-PCR和Western blot法,观察AGE-HSA对DCs VCAM-1 mRNA和蛋白表达的影响,同时检测培养液上清中IL-12和IL-18的浓度。结果:与空白对照相比,AGE-HSA可上调DCs VCAM-1 mRNA和蛋白的表达(P<0.05),并且明显促进了DCs IL-12和IL-18的分泌(P<0.05)。AGE-HSA干预组与空白对照组相比差异无统计学意义(P>0.05)。结论:AGEs能够上调DCs VCAM-1的表达,并且促进DCs IL-12和IL-18的分泌,这可能是糖尿病通过DCs促进动脉粥样硬化发生的重要机制之一。

Objective：To investigate the effect of advanced glycosylation end products（AGEs） on the expression of vascular cell adhesion molecule-1（VCAM-1） in human monocyte-derived dendritic cells（DCs）.Methods：Monocytes were purified（over 98%） using anti-CD14 microbeads.After cultured with DC Cellgro medium containing recombinated human granulocyte-macrophage colony stimulating factor（rhGM-CSF,100 μg/L） and recombinated human interleukin-4（rhIL-4,20 μg/L） for 8 days,monocytes were derived into immature DCs.Then they were exposed to AGE-HSA（0,200 μg/ml） for 24 h.Expression of VCAM-1 was semi-quantified by RT-PCR and Western-blot.At the same time,supernatants were collected and IL-12,IL-18 were analyzed by ELISA.Results：mRNA and protein of VCAM-1 incubated by 200 μg/ml AGE-HSA was higher than those of control at 24 h.Treatment of DCs with AGE-HSA resulted in nearly two times of the expression of VCAM-1（P 0.05）.The concentration of IL-12 and IL-18 was significantly higher than that of control（P 0.05）.Conclusion：AGEs can up-regulate the expression of VCAM-1 and induce the secretion of IL-12 and IL-18 in DCs.These findings may provide insight into the effect of DCs in processes of atherosclerosis.