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为找到一种判断泡沫细胞与非泡沫细胞的简便而较为实用的方法,本文采用正己烷-异丙醇从U937单核细胞内提取胆固醇,并用醇溶性氢氧化钾除去其中的甘油三酯成分,用三氯乙酸去其中的蛋白成分。以异丙醇:正己烷:乙腈为溶剂洗脱系统,采用疏水性较弱的核酸分离柱,进行高效液相色谱分析,分离纯化胆固醇和胆固醇酯成分,并采用L-B(LiebermanBurchard)显色法结合波谱分析法对所胆固醇峰进行鉴定。结果表明胆固醇含量在0.05~1g/L之间与其峰面积有较好的线性关系,其最低检出量为4mg/g细胞蛋白(100mg细胞蛋白相当于1×108个细胞),通过定性与定量分析,结果表明,U937单核细胞株经氧化型低密度脂蛋白处理48h后,其胞内的胆固醇酯占胞内总胆固醇60%以上。
In order to find out a simple and practical method for judging foam cells and non-foam cells, cholesterol is extracted from U937 mononuclear cells by n-hexane-isopropanol and the triglyceride components are removed by alcohol-soluble potassium hydroxide. With trichloroacetic acid to which the protein composition. Using isopropanol: n-hexane: acetonitrile as solvent elution system, the hydrophobic hydrophobic nucleic acid separation column was used for high performance liquid chromatography, and the cholesterol and cholesterol ester components were separated and purified by L-B (Lieberman Burchard) Law combined with spectroscopy to identify the peak of cholesterol. The results showed that the content of cholesterol in the range of 0.05 ~ 1g / L had a good linear relationship with the peak area, the minimum detectable amount of 4mg / g cell protein (100mg cell protein equivalent to 1 × 108 cells), by qualitative The results showed that the intracellular cholesterol ester accounted for more than 60% of intracellular total cholesterol after U937 mononuclear cell line was treated with oxidized low density lipoprotein for 48h.