脂蛋白对培养巨噬细胞表达噬巨细胞移动抑制因子的实验研究  

Effects of different lipid on MIF mRNA and protein expression in cultural macrophage

作　　者：林秋雄[1] 单志新[1] 余细勇[1] 杨敏[1] 林曙光[1] Hui-yao LAN 

机构地区：[1]广东省人民医院医学研究中心,广东省冠心病研究重点实验室 [2]Department of Medicine, Baylor College of Medicine, Houston, Texas, USA

出　　处：《中国临床药理学与治疗学》2005年第3期329-332,共4页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：国家自然科学基金项目 (№ 3 0 3 0 0 42 1) ;广东省自然科学基金团队项目 (№ 0 15 0 15 )

摘　　要：目的:探讨不同脂蛋白对培养的人巨噬细胞表达巨噬细胞移动抑制因子基因和蛋白水平的影响。方法:2 8SC人巨噬细胞株用含有1×10 5U·L-1青霉素、10 0mg·L-1链霉素和10 %FBS的RPMI 16 40培养基于37℃,5 %的CO2 中培养。以每孔1×10 4个细胞接种于6孔板中,每孔2ml培养液,加入终浓度为15 0mg·L-1的不同脂蛋白,于37℃共同孵育2 4h收集细胞和培养介质。采用RT PCR和ELISA分别测定MIFmRNA和蛋白表达水平。结果:巨噬细胞有MIF表达,低密度脂蛋白、氧化型低密度脂蛋白、胆固醇和甘油三脂组诱导巨噬细胞MIFmRNA和蛋白表达水平,试验组较正常对照组升高(P <0 .0 5 ) ;而高密度脂蛋白和极低密度脂蛋白组诱导巨噬细胞MIFmRNA和蛋白表达水平,试验组与正常对照组变化相近(P >0 .0 5 )。结论:不同脂质对巨噬细胞MIFmRNA和蛋白表达作用不同。MIF可能参与了低密度脂蛋白、氧化型低密度脂蛋白、胆固醇和甘油三脂所致的动脉粥样硬化进程。AIM: To investigate the expression of MIF mRNA and protein in cultural human macrophages induced by different lipoproteins. METHODS: 28sc human macrophage cell lines were grown in RPMI-1640 medium containing 1×105 U·L -1 penicillin, 100 mg·L -1 streptomycin, 10% FBS at 37 ℃ and 5% CO_2. The concentrations of cells were adjusted as 10×104 ml -1, and then macrophages were inoculated in 6 wells plate. Different lipoproteins with a final concentration as 150 mg·L -1 were added respectively in each well. After culture 24 h together, the expression of MIF mRNA and protein were detected by RT-PCR and ELISA methods. RESULTS: The MIF mRNA and protein were expressed in macrophage. LDL, Ox-LDL, cholesterol and TG can significantly increased the expression of MIF mRNA and protein in macrophage compared with the control group(P< 0.05). However, HDL and VLDL showed no effect on the expression of MIF mRNA and protein in macrophage(P> 0.05). CONCLUSION: Different lipid including LDL, Ox-LDL, cholesterol and TG can up-regulate MIF mRNA and protein expression in macrophage. It suggests that MIF may involve in the progress of atherosclerosis.

关 键 词：脂蛋白类 巨噬细胞移动抑制因子 动脉粥样硬化 细胞培养 逆转录聚合酶链反应 酶联免疫分析 

分 类 号：R966[医药卫生—药理学]