糖基化终产物对人单核细胞源树突状细胞清道夫受体A表达的影响及其机制的研究  

Effect and mechanism of advanced glycosylation end products on the expression of scavenger receptor A in human monocyte-derived dendritic cells

作　　者：贾庆哲[1] 葛均波[1] 梁春[2] 罗育坤[1] 黄东[1] 王克强[1] 陈灏珠[1] 

机构地区：[1]上海市心血管病研究所复旦大学附属中山医院心内科,200032 [2]第二军医大学长征医院心内科

出　　处：《中华心血管病杂志》2004年第10期888-892,共5页Chinese Journal of Cardiology

基　　金：国家重点基础研究发展规划资助项目 (G2 0 0 0 0 5 690 3 ) ;上海市科委资助项目 ( 0 2JC14 0 2 6)

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 (MDCs)清道夫受体A(SR A)表达的影响及其机制。方法　用免疫磁珠分离人外周血CD14 + 单核细胞 ,经含重组人粒 巨噬细胞集落刺激因子 (rhGM CSF ,10 0ng/ml)和重组人白细胞介素 4 (rhIL 4 ,5 0ng/ml)的RPMI16 4 0培养 ,使其分化为MDCs,采用RT PCR和Western Blot法 ,分别观察糖基化 白蛋白 (AGE BSA)不同蛋白浓度 (0、5 0、10 0、2 0 0、30 0 μg/ml)和不同时间 (0、6、12、2 4、36h)干预 ,以及酪氨酸蛋白激酶抑制剂金雀异黄素 (genistein)干预后 ,MDCsSR A基因和蛋白的表达。结果　与空白对照相比 ,蛋白浓度为 5 0 μg/ml和 10 0 μg/ml干预 2 4h即可分别上调SR AmRNA和蛋白的表达 (P <0 0 5 ) ,2 0 0 μg/ml时达峰值 (P <0 0 1) ,在干预的不同时间组 ,12h和 2 4h可分别上调SR AmRNA和蛋白的表达 (P<0 0 5 ) ,36h达峰值 (P <0 0 1) ,呈明显的浓度和时间依赖性 ,genistein能够完全抑制其作用。 结论AGEs能够上调DCsSR A的表达 ,是与其激活酪氨酸蛋白激酶有关 ,这可能是DCs参与动脉粥样硬化发生的机制之一。Objective To investigate the effect of advanced glycosylation end products (AGEs) on the e xpression of scavenger receptor A (SR-A) in human monocyte-derived dendritic c ells (MDCs) and it′s mechanism. Methods Monocytes were puri fied (over 98%) using Anti-CD14+ microbeads. After eight days′ culture in R PMI1640 medium containing recombinated human granulocyte-macrophage colony stim ulating factor (rhGM-CSF,100 ng/ml) and recombinated human interleukin-4 (rhI L-4,50 ng/ml), immature MDCs were derived. They were exposed to AGE-BSA (pro tein concentration of 0, 50, 100, 200, and 300 μg/ml respectively) for 24 hour s, and exposed to AGE-BSA (protein concentration of 200 μg/ml) for 0, 6, 12, 24, and 36 hours respectively. Expression of SR-A was semi-quantified by RT-P CR and Western-Blot. Finally the cells were intervened by genistein to investi gate the mechanism of the gene and protein expression of MDCs SR-A. Res ults The mRNA of SR-A incubated by 50 μg/ml AGE-BSA was higher than that of control and the protein of SR-A incubated by 100 μg/ml AGE-BSA was h igher than that of control at 24 h (P<0.05). It reached the peak at 200 μg/ml AGE-BSA(P<0.01). After intervention of AGE-BSA for periods of 1 2, 24 ,36 hours, the SR-A expression was increased markedly, reaching the peak at 36 hours (P<0.01). Genistein could inhibit the expression of SR-A cau sed by AGE-BSA completely. Conclusion AGEs can up-regulate t he expression of SR-A in DCs. Its mechanism might be related to AGEs activating tyrosine protein kinase. This finding might provide an insight into the effect of DCs on the formation of atherosclerosis.

关 键 词：表达 干预 人单核细胞 糖基化终产物 树突状细胞 上调 不同时间 清道夫受体A 蛋白浓度 RNA 

分 类 号：R543[医药卫生—心血管疾病]