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目的:探究低密度脂蛋白(LDL)与脂质筏之间的关系。方法:运用荧光漂白恢复技术(FRAP),对LDL刺激的细胞和未经LDL刺激的细胞进行观测;采用密度梯度离心的方法分离脂质筏,对各层中胆固醇含量进行测定,通过Western Blot对脂质筏标志性蛋白flotillin-1,caveolin-1进行分析;β-环糊精孵育后,扫描电子显微镜观测细胞形态。结果:经LDL刺激的Raw264.7细胞表现出荧光漂白恢复时间延长;脂质筏区的胆固醇含量增多,脂质筏区标志性蛋白flotillin-1,caveolin-1增加;β-环糊精孵育后细胞形态发生改变,细胞胆固醇含量降低。结论:LDL能诱导细胞膜上脂质筏的聚集。
Objective: To explore the relationship between low density lipoprotein (LDL) and lipid rafts. Methods: Fluorescence bleaching recovery (FRAP) was used to observe the effects of LDL-stimulated cells and non-LDL stimulated cells. Lipid rafts were separated by density gradient centrifugation, and the cholesterol content in each layer was determined. Western Blot Lipid raft landmark protein flotillin-1, caveolin-1 were analyzed; β-cyclodextrin incubation, scanning electron microscopy cell morphology. RESULTS: Raw264.7 cells stimulated by LDL showed prolonged recovery time of fluorescence bleaching, cholesterol content in lipid raft region increased, flotillin-1 and caveolin-1, which are markers in lipid raft region, increased. After incubation with β-cyclodextrin Cell morphology changes, decreased cellular cholesterol levels. Conclusion: LDL can induce the accumulation of lipid rafts on the cell membrane.