重庆医学2025，Vol.54Issue(3)：580-587,8.DOI:10.3969/j.issn.1671-8348.2025.03.004
外被体蛋白Ⅰ参与载脂蛋白A-1介导泡沫细胞胆固醇流出的作用研究
Study on the role of coatomer protein Ⅰ in apolipoprotein A-1-mediated cholesterol efflux from foam cells
洪伟涛 1张飞龙 1黄婕 1梁钰敏 1陈耿基 1陈晓佳 1马卫列 1丁航 1张志珍1
作者信息
- 1. 广东医科大学基础医学院,广东 东莞 523808
- 折叠
摘要
Abstract
Objective To establish a human monocytic THP-1 macrophage-derived foam cell model and analyze the role of the α subunit of coatomer protein Ⅰ(α-COP)in apolipoprotein A-1(apoA-1)-mediated cholesterol efflux from foam cells.Methods THP-1 cells were induced to adhere using PMA(PMA group),followed by treatment with acetylated low-density lipoprotein(Ac-LDL)to generate macrophage-derived foam cells(Ac-LDL group).Subsequent incubation with apoA-1 formed the apoA-1 group.Cholesterol efflux rates mediated by apoA-1 and intracellular lipid accumulation were quantified through liquid scintillation counting and oil red O staining.The expression patterns of α-COP were systematically analyzed using quantitative real-time PCR(qPCR),Western blotting,and laser scanning confocal microscopy.THP-1 macrophage-derived foam cells mediated by apoA-1 were transduced with scrambled shRNA(Scr group)or α-COP-specific shRNA lentivirus(α-COP shRNA group)to assess the effects of α-COP knockdown on cholesterol efflux efficiency and total cellular cholesterol content.Subcellular localization of adipopoilin(ADFP)and cholesterol probe Fil-ipin Ⅲ staining patterns were visualized via laser scanning confocal microscopy.Bioinformatics analysis of α-COP expression profiles in carotid atherosclerotic plaques was performed using datasets from the GEO data-base.Results Compared with the Ac-LDL group,the cholesterol efflux rate in the apoA-1 group was signifi-cantly increased[(9.77±0.79)%vs.(2.74±0.37)%,P＜0.001].Oil red O staining demonstrated reduced lipid accumulation in foam cells of the apoA-1 group compared with the Ac-LDL group.The relative mRNA expression level of α-COP in the apoA-1 group was significantly higher than that in the Ac-LDL group(P＜0.001),with corresponding elevation in protein expression(P＜0.001).Fluorescence intensity analysis re-vealed increased mean fluorescence intensity of α-COP in the apoA-1 group compared with the Ac-LDL group.Following α-COP knockdown by shRNA,the apoA-1-mediated cholesterol efflux rate was significantly de-creased(P＜0.05)and total intracellular cholesterol levels were increased(P＜0.001)in the α-COP shRNA group compared with the Scr group.The α-COP shRNA group exhibited enhanced red fluorescence intensity of ADFP(P＜0.05)and increased blue fluorescence intensity of Filipin Ⅲ(P＜0.01)compared with the Scr group.Database analysis indicated that α-COP mRNA expression was lower in carotid atherosclerotic plaques than in normal arterial tissues,with reduced expression in advanced-stage plaques compared with early-stage plaques(P＜0.05).Conclusion α-COP participates in apoA-1-mediated cholesterol efflux from foam cells,and interference with α-COP expression results in reduced cholesterol efflux and increased intracellular lipid accumulation.关键词
外被体蛋白Ⅰ/载脂蛋白A-1/泡沫细胞/胆固醇流出Key words
coatomer protein Ⅰ/apolipoprotein A-1/foam cell/cholesterol efflux分类
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洪伟涛,张飞龙,黄婕,梁钰敏,陈耿基,陈晓佳,马卫列,丁航,张志珍..外被体蛋白Ⅰ参与载脂蛋白A-1介导泡沫细胞胆固醇流出的作用研究[J].重庆医学,2025,54(3):580-587,8.
