别嘌呤醇抑制巨噬细胞脂质蓄积的作用机制  

Study on the Mechanism of Allopurinol Inhibiting the Lipid Accumulation of Macrophages

作　　者：郑伟[1] 陈欣[1] 李亚新[1] 李珊[1,2] 

机构地区：[1]湖北医药学院附属东风医院综合医疗科,湖北省十堰市442000 [2]湖北医药学院基础医学院生物化学教研室,湖北省十堰市442000

出　　处：《中国动脉硬化杂志》2016年第4期355-360,共6页Chinese Journal of Arteriosclerosis

基　　金：湖北省教育厅科学技术研究计划优秀中青年人才项目(Q20132105);湖北省教育厅科学技术研究计划重点项目(D20142106)

摘　　要：目的采用氧化型低密度脂白(ox-LDL)构建小鼠巨噬细胞泡沫化模型,探讨别嘌呤醇对泡沫化进程中脂质蓄积的影响及其机制。方法体外培养小鼠单核巨噬细胞株RAW264.7,用ox-LDL或Dil-ox-LDL孵育细胞构建泡沫化模型。用不同浓度别嘌呤醇处理细胞,MTT法筛选出合适的实验浓度。将合适浓度的别嘌呤醇作用于细胞,在共聚焦荧光显微镜下观察Dil-ox-LDL孵育后RAW264.7细胞内脂质蓄积情况;酶学终点法定量检测细胞内总胆固醇的变化;半定量﹑荧光定量RT-PCR法和蛋白免疫印迹法分别检测细胞中LXRα和ABCA1 mRNA和蛋白的表达水平。结果与ox-LDL诱导的泡沫化模型组相比,别嘌呤醇组脂质蓄积显著下降,总胆固醇含量明显下调。别嘌呤醇在mRNA和蛋白水平上引起LXRα和ABCA1的高表达。结论别嘌呤醇具有抑制ox-LDL诱导小鼠巨噬细胞泡沫化进程的作用,并通过上调LXRα-ABCA1途径来调节细胞内胆固醇的含量。Aim To construct the mouse macrophage foam model by using oxidized low density lipoprotein（ oxLDL）;To investigate the effect of allopurinol on lipid accumulation in macrophage foam process and its mechanism.Methods Mouse macrophage cell line RAW264.7 was cultured in vitro,and foam model was constructed by using ox-LDL or Dil-ox-LDL incubating macrophage cells.Macrophage cells were treated with different concentrations of allopurinol,and MTT method was used to screen the suitable experimental concentration.Macrophage cells were treated with suitable concentration of allopurinol.After Dil-ox-LDL incubation,the lipid accumulation in RAW264.7 cells was observed under confocal fluorescence microscopy.The change of total cholesterol in the cells was detected by enzymatic end point method.Semi-quantitative,fluorescence quantitative RT-PCR and Western blot were used to detect the expressions of liver X receptor α（ LXRα）,ATP-binding cassette transporter A1（ ABCA1） mRNA and protein in cells.Results Compared with the ox-LDL induced foam model group,the lipid accumulation and total cholesterol content were significantly decreased in RAW264.7 cells of the allopurinol group.Allopurinol could cause the high expressions of LXRα,ABCA1 mRNA and protein.Conclusion Allopurinol can restrain mouse macrophage foam process induced by ox-LDL,and regulate intracellular cholesterol content through the up-regulation of LXRα-ABCA1 pathway.

关 键 词：别嘌呤醇 巨噬细胞 脂质蓄积 肝X受体Α 三磷酸腺苷结合盒转运体A1 

分 类 号：R363[医药卫生—病理学]