HPLC法测定RAW264．7源性泡沫细胞中胆固醇酯的含量

Content Determination of Cholesterol Esters in Foam Cells Derived from RAW264.7 by HPLC

目的:建立测定细胞内胆固醇及胆固醇酯含量的高效液相色谱法(HPLC),以精确鉴定泡沫细胞含量,为研究细胞内胆固醇的变化提供技术平台。方法:应用HPLC法测定氧化低密度脂蛋白(OX-LDL)和不同浓度的普伐他汀处理RAW264.7鼠源单核细胞源性巨噬细胞中胆固醇酯的含量。结果:OX-LDL可显著增加细胞对酯质的摄取量(P<0.01),而普伐他汀能通过降低细胞内胆固醇水平而减轻细胞泡沫化程度,且呈浓度、时间依赖性(P<0.05,P<0.01)。结论:本研究建立的测定细胞内胆固醇及胆固醇酯含量的HPLC法可用来鉴定泡沫细胞的泡沫化程度。

OBJECTIVE： To establish an HPLC method for the determination of levels of intracellular cholesterol and cholesterol esters in foam cells derived from RAW264.7 murine macrophages so as to obtain a precise measure of foam cells and provide a technical platform for the study of the intracellular cholesterol content change. METHODS： HPLC method was applied to determine the content of cholesterol esters in the foam cells treated with OX- LDL and different concentration of pravastatin. RESULTS： OX- LDL could significantly enhance cellular intake of lipids （P〈0.01） while pravastatin could lessen the formation of foam cells in a dose- dependent and time- dependent manner （P 〈 0.05, P 〈 0.01） . CONCLUSION： The established HPLC method can be used to determine the levels of intracellular cholesterol and cholesterol esters and detect the formation of cellular foam cells.