摘要
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。
Aim To determine cholesterol and cholesteryl ester of U937 monocyte within it's foaming procedure,and to obtained a simple and useful method to identify foamcell.Methods Cholesterol and cholesteryl ester was ex-tracted from U937 monocyte by hexane-isopropanol,and it's triglycerides was hydrolyzed by ethanolic KOH,and it's protein contents was removed by trichloroacetic acid.Father separation of cholesterol and cholesteryl ester was accompolished by high per-formance liquid chromatography(HPLC),using an eluent of isopropanol-n-heptane-acetonitrice and a Gen-Pak FAX column(with a little hydrophobic prop-erty,using for nucleic acid separation)and it's choles-terol peak was identified by L-B(Lieberman burchard)reaction and spectrum index analysis.Results Cholesterol concentration has a high linear relationship with it's peak area within 0.05-1.0g/L,the limits of detection of cholesterol is 4mg/g cell pro-tein(100mg cell protein equivalent to 1×108 cells).The cholestelyl ester inside U937 amount to 62% to total cholesterol after cholesterol-loaded for 48 hour.Conclusion This method is useful and simple to i-dentify foam cell.
出处
《中国动脉硬化杂志》
CAS
CSCD
1998年第4期317-320,共4页
Chinese Journal of Arteriosclerosis
基金
湖南省中青年基金
关键词
高效液相色谱
U937单核细胞株
胆固醇
胆固醇酯
High performance liquid chromatography
U937 monocyte
Cholesteryl
Cholesteryl ester
