氧化型低密度脂蛋白对RAW264．7细胞Siglec-1表达及分泌细胞因子的影响  

Increased expression of Siglec-1 and cytokines by stimulation of oxidized low-density lipoprotein on RAW264.7 cells

作　　者：熊怡淞[1] 朱烨[1] 陆慧琦[1] 李畅[1] 韩志君[1] 仲人前[1] 

机构地区：[1]第二军医大学长征医院实验诊断科,全军医学免疫诊断中心,全军临床免疫重点实验室,上海200003

出　　处：《中华微生物学和免疫学杂志》2010年第1期51-55,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家高科技研究发展计划（863计划）（2006AA02Z496）;上海市科学技术委员会优秀学科带头人基金（07XD14013）

摘　　要：目的通过氧化型低密度脂蛋白（oxidized low—density lipoprotein，ox-LDL）活化巨噬细胞系对Siglec-1（sialic acid—binding immunoglobulin-like lectin 1，唾液酸黏附素）表达及细胞因子分泌的影响探讨对动脉粥样硬化（atherosclerosis，AS）发病的可能作用。方法制备ox—LDL，并用不同浓度ox-LDL刺激小鼠巨噬细胞株RAW264．7，48h后收集细胞和上清，分别用流式细胞仪和RT—PCR检测不同浓度ox-LDL刺激后细胞表面Siglec-1蛋白和基因表达水平，并测定培养上清液中巨噬细胞炎性蛋白-1α（MIP-1α）、单核细胞趋化蛋白-1（MCP-1）和IL-8的浓度。结果与对照组相比，ox—LDL能刺激RAW264．7细胞表面Siglec-1蛋白和mRNA表达升高（P〈0．01），培养上清液中MIP-1α、MCP-1和IL-8的表达升高（P〈0．01），并呈浓度依赖性。结论ox—LDL可呈剂量依赖性刺激RAW264．7巨噬细胞诱导Siglec-1表达及分泌炎性细胞因子增加，在动脉粥样硬化的发生发展中可能起一定作用。Objective To explore the role of sialic acid-binding immunoglobulin-like lectin-one （Siglec-1） in the process of atheroselerotic inflammation induced by oxidized low-density lipoprotein （ox- LDL）. Methods Ox-LDL was synthesized by oxidization of native LDL and different concentration of ox- LDL was added to the culture medium of RAW264.7. Forty-eight hours later, cells and supernatants were collected separately. The expression of Siglec-1 protein and mRNA were measured by flow cytometry（FCM） and real-time quantitative RT-PCR, respectively. The levels of monocyte chemotactic protein-1 （MCP-1）, macrophage inflammatory protein-1α（MIP-1α） and IL-8 in supernatants were determined by ELISA. Resuits By the stimulation of ox-LDL, Siglec-1 protein and mRNA on RAW264.7 cells were significantly increased, meanwhile, the cytokines levels in culture supernatants were significantly higher than that in the control group. And both Siglec-1 expression and cytokine secretion were ox-LDL dose-dependent. Conclusion Ox-LDL can increase Siglec-1 protein and mRNA expression and some inflammatory cytokines secretion on RAW264.7 cells in a dose-dependent manner. Manifested by enhanced Siglec-1 expression, the activated macrophages may take a part in the development and progression of atherosclerosis.

关 键 词：动脉粥样硬化 氧化型低密度脂蛋白 SIGLEC-1 RAW264.7 

分 类 号：R285.1[医药卫生—中药学] R543.5[医药卫生—中医学]