一种快速分析细胞内脂质沉积的方法

A Method for Rapid Measurement of Intracellular Lipid Accumulation

目的:建立快速分析细胞内脂质沉积的方法。方法:以油酸刺激人肝癌细胞形成脂质沉积,利用BODIPY 493/503染色细胞内脂滴,在Matlab环境下以荧光图像分析技术分析脂滴荧光强度;采用蛋白印迹法分析脂质沉积标志蛋白——脂滴表面蛋白Adipophilin的蛋白水平,以及采用实时荧光定量PCR法检测Adipophilin的基因表达水平,以验证荧光图像分析技术的可靠性。结果:利用荧光图像分析技术建立了快速定量脂质沉积的方法,结果显示该方法下400μmol/L油酸显著提高细胞内脂质含量达1.41倍,与标志蛋白Adipophilin的蛋白水平增高2.07倍,基因表达水平增高1.53倍一致。结论:该可快速定量细胞内脂质沉积水平,且结果可靠。

Objective： To propose a fast and reliable method for the determination of intracellular lipid accumulation. Method： Lipid accumulation in Hep G2 cells were induced by oleic acid. Lipid droplets were stained with BODIPY 493 /503 and measured with fluorescent image processing using Matlab or Image J. The images results were compared to those of Adipophilin,a widely served marker for lipid accumulation,which were determined from protein and mRNA levels by Western blotting and q PCR. Result： By processing fluorescent images using BODIPY 493 /503 staining for lipid droplets and comparing data outputs with Matlab and Image J,a method using Matlab as image processing platform was established for the determination of intracellular lipid accumulation; a 1. 41- fold increase in oleic acid- induced lipid accumulation was detected by fluorescent image processing,which were in accordance with the markedly enhancement in Adipophilin protein levels（ 2. 07- fold） and mRNA levels（ 1. 53- fold）. Conclusion： The method combining fluorescent staining and image processing shall help analyze the intracellular lipid accumulation both rapidly and accurately.