【摘要】 目的:测定血管内皮细胞膜小凹微区域中胆固醇含量。方法:建立用反相高效液相色谱-二极管阵列检测器测定细胞膜小凹微区域中胆固醇的方法。细胞内皮小凹微区域分离组分,经乙醚与石油醚的混合二次萃取后,经氮气吹干浓缩,在Symmetry Shield RP-18柱上经流动相乙腈与异丙醇(8∶1,v/v)洗脱,检测六份不同处理细胞膜小凹微区域的胆固醇。结果:实验方法中胆固醇的峰面积与质量浓度的线性关系良好,线性相关系数为0.992 182,平均回收率>95.6%。结论:用n-3多不饱和脂肪酸处理后和未加脂肪酸处理的小凹膜微区域组分中的胆固醇含量无明显变化,而甲基-β-环糊精可明显降低细胞膜小凹微区域的胆固醇含量。更多还原

【Abstract】 Objective:To determine cholesterol content in caveolae microdomains of HUVEC cells.Methods:A reversedphase HPLC-DAD method was developed to determine cholesterol in caveolae microdomains.Caveolae fractions were extracted twice with diethyl ether-petroleum ether,dried under nitrogen,and seperated with HPLC.A Symmetry Shield RP-18 column was selected and the mobile phase comprised of acetonitrile-isopropanol(8∶1).Results:A linearity was obtained with a good coefficient(r=0.992 182).The average recovery was higher than 95.6%.Conclusion:n-3 polyunsaturated fatty acid(n-3 PUFA)treatment does not influence the cholesterol content in caveolae compared with untreated cells.However,methy1-β-cyclodextran can significantly decrease cholesterol content in caveolae microdomains.更多还原