中国动脉硬化杂志2026，Vol.34Issue(4)：344-351,8.DOI:10.20039/j.cnki.1007-3949.2026.04.008
获得富含胆固醇脂蛋白颗粒的单步密度梯度离心法
Single-step density gradient centrifugation method for obtaining cholesterol-rich lipo-protein particles
摘要
Abstract
Aim To establish a single-step density gradient centrifugation method for isolating cholesterol-rich non-high density lipoprotein(CR-nHDL)particles and investigate the effect of oxidized CR-nHDL on foam cell formation.Methods A single-step density gradient centrifugation method using ioversol as the density medium was established to iso-late CR-nHDL with a density of less than 1.008 g/mL from postprandial plasma of healthy volunteers.The density of the fraction was measured,and the cholesterol content was detected enzymatically.The lipoprotein content in the particles was analyzed by Western blot and proteomics.The isolated CR-nHDL was oxidized and co-cultured with mouse bone mar-row-derived macrophages(BMDM),and its effect was compared with that of oxidized low density lipoprotein(ox-LDL).The ability of oxidized CR-nHDL to induce macrophage foam cell formation was assessed by measuring intracellular lipid ac-cumulation.Results This method successfully isolated CR-nHDL with a density of less than 1.008 g/mL.Western blot results showed that this fraction contained apolipoprotein B100(ApoB100)and apolipoprotein B48(ApoB48)but not apolipoprotein AⅠ(ApoAⅠ).The results of proteomic analysis showed that there was no significant difference in the major apolipoprotein components between CR-nHDL prepared by ioversol method and low density lipoprotein(LDL)pre-pared by classical two-step method;lipidomics analysis results showed that the cholesterol ester content in the CR-nHDL prepared by the ioversol method was significantly higher than that in the LDL prepared by the classical method,which sug-gested that this method was more efficient in enriching cholesterol-containing lipoprotein particles,and the oxidized CR-nHDL obtained from this method could promote lipid accumulation in macrophage derived foam cells more than ox-LDL.Conclusion The single-step density gradient centrifugation of ioversol can quickly isolate CR-nHDL,and its oxidized form has a stronger ability to promote macrophage foam cell formation.关键词
单步密度梯度离心法/富含胆固醇的非高密度脂蛋白/泡沫细胞/碘佛醇/动脉粥样硬化Key words
single-step density gradient centrifugation method/cholesterol-rich non-high density lipoprotein cho-lesterol/foam cell/ioversol/atherosclerosis分类
医药卫生引用本文复制引用
刘意秋,张继国,耿长鑫,王蕾,张昭强,张亚,谢丽丽,杨泽,张科,于杨..获得富含胆固醇脂蛋白颗粒的单步密度梯度离心法[J].中国动脉硬化杂志,2026,34(4):344-351,8.基金项目
国家自然科学基金面上项目(81970385) （81970385）
山东省自然科学基金项目(ZR2023MH361、ZR2022MH202) （ZR2023MH361、ZR2022MH202）
山东省医药卫生科技发展计划项目(202002040923) （202002040923）
泰安市科技发展计划引导项目(2020NS232) （2020NS232）
泰安市发展创新发展项目(2023NS379) （2023NS379）
泰安市农业和社会发展领域科技创新发展项目(2024NS463) （2024NS463）
