Lü Zhan, Gou Lian-ping, Chen Ling, Xie Bin, Qin Jian
Department of Cardiology, Affiliated Hospital of North Sichuan Medical College, Sichuan Nanchong 637000, China.
Zhonghua Nei Ke Za Zhi. 2010 Aug;49(8):696-9.
This study was designed to explore the function of ATP binding cassette transporter 1 (ABCA1) and Apolipoprotein A-I (ApoA-I) in cholesterol reverse transportation (RCT), the influence of lovastatin and rosiglitazone on the concentration of cholesterol (CHO) in THP-1 (human monocytic leukemia cell line) derived foam cells.
LDL from healthy volunteers was obtained by density-gradient ultracentrifugation and was oxidized by incubation with Cu2+ and ox-LDL was identified.Macrophages were induced from THP-1 cell by phorbol ester (PMA). Models of foam cells were built by incubating macrophages with oxLDL. The effect of lovastatin and rosiglitazone on ABCA1 protein expression in THP-1 cell line derived macrophage were detected by western blot. Foam cells were divided into 9 groups: control, ApoA-I, lovastatin, rosiglitazone lovastatin+ApoA-I, rosiglitazone+ApoA-I, ABCA1 monoclonal antibody pretreatment+ApoA-I, ABCA1 monoclonal antibody pretreatment+lovastatin+ApoA-I, ABCA1 monoclonal antibody pretreatment+rosiglitazone+ApoA-I. The concentration of intracellular CHO in each group was detected by using cholesterol kit.
As compared with control group, there are no big differences of CHO concentration within the cell of group lovastatin, rosiglitazone, and each ABCA1 monoclonal antibody pretreatment group (P>0.05), but the CHO concentration within the cells of group ApoA-I, lovastatin+ApoA-I, rosiglitazone+ApoA-I decreased obviously as compared with the control (P<0.05), and CHO concentration in group rosiglitazone+ApoA-I have a further decrease than the former two groups (P<0.05).
CHO concentration can be decreased in foam cells by cooperation of ABCA1 and ApoA-I mediate cholesterol efflux. Rosiglitazone can enhance this procedure in THP-1 macrophages derived foam cells which means that they can promote ABCA1 mediated cholesterol reverse transportation through improve ABCA1 protein expression.
本研究旨在探讨ATP结合盒转运蛋白1（ABCA1）和载脂蛋白A-I（ApoA-I）在胆固醇逆向转运（RCT）中的作用，以及洛伐他汀和罗格列酮对THP-1（人单核细胞白血病细胞系）源性泡沫细胞中胆固醇（CHO）浓度的影响。
通过密度梯度超速离心法从健康志愿者中获取低密度脂蛋白（LDL），并与Cu2+孵育使其氧化，鉴定氧化型低密度脂蛋白（ox-LDL）。用佛波酯（PMA）诱导THP-1细胞分化为巨噬细胞。将巨噬细胞与ox-LDL孵育构建泡沫细胞模型。采用蛋白质免疫印迹法检测洛伐他汀和罗格列酮对THP-1细胞系源性巨噬细胞中ABCA1蛋白表达的影响。将泡沫细胞分为9组：对照组、ApoA-I组、洛伐他汀组、罗格列酮组、洛伐他汀+ApoA-I组、罗格列酮+ApoA-I组、ABCA1单克隆抗体预处理+ApoA-I组、ABCA1单克隆抗体预处理+洛伐他汀+ApoA-I组、ABCA1单克隆抗体预处理+罗格列酮+ApoA-I组。使用胆固醇试剂盒检测每组细胞内CHO的浓度。
与对照组相比，洛伐他汀组、罗格列酮组及各ABCA1单克隆抗体预处理组细胞内CHO浓度差异无统计学意义（P>0.05），但ApoA-I组、洛伐他汀+ApoA-I组、罗格列酮+ApoA-I组细胞内CHO浓度较对照组明显降低（P<0.05），且罗格列酮+ApoA-I组CHO浓度低于前两组（P<0.05）。
ABCA1和ApoA-I协同介导胆固醇流出可降低泡沫细胞内CHO浓度。罗格列酮可增强THP-1巨噬细胞源性泡沫细胞中的这一过程，即通过提高ABCA1蛋白表达促进ABCA1介导的胆固醇逆向转运。
