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
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江苏省高校自然科学基金(06KJB320006)


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

    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) 100 μg/L和重组人白细胞介素-4(rhIL-4) 20 μg/L的RPMI1640培养,使其分化为DCs,加入糖基化人血清白蛋白(AGE-HSA) 200 μg/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-

    Abstract:

    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.

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贾庆哲,朱铁兵,王连生,李春坚,陶正贤,杨志健.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[J].南京医科大学学报(自然科学版),2012,(9):1211-1214

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