载脂蛋白A-Ⅰ通过PKA信号途径影响ABCA1的表达与功能
DOI:
CSTR:
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

基金项目:

国家自然科学基金(30470720),中国博士后科学基金(2005037157)和湖南省自然科学基金(06jj5058)资助项目.


Effect of Apolipoprotein A-Ⅰ on Expression and Function of ATP-binding Cassette Transporter A1 Through PKA Signaling
Author:
Affiliation:

Fund Project:

This work was supported by the grants from The National Natural Sciences Foundation of China (30470720), Post-doctor Sciences Foundation of China (2005037157) and Hunan Provincial Natural Sciences Foundation of China (06jj5058).

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    以THP-1巨噬细胞源性泡沫细胞为研究对象,观察载脂蛋白A-Ⅰ与三磷酸腺苷结合盒转运体A1(ATP binding cassette transporter A1,ABCA1)的相互作用,并探讨它们相互作用的机制,以便了解载脂蛋白A-Ⅰ和ABCA1在动脉粥样硬化发生发展中的作用. THP-1巨噬细胞源性泡沫细胞经各种因素处理后,采用油红“O”染色, 观察细胞内的脂滴,运用液体闪烁计数器检测细胞内胆固醇流出,高效液相色谱分析细胞内总胆固醇、游离胆固醇和胆固醇酯含量, 用逆转录-聚合酶链反应和蛋白质印迹分析法分别检测ABCA1 mRNA与ABCA1蛋白质的水平. 实验结果显示,载脂蛋白A-Ⅰ和腺苷酸环化酶激动剂Forskolin(FRK)引起THP-1巨噬细胞源性泡沫细胞总胆固醇、游离胆固醇与胆固醇酯减少,而腺苷酸环化酶抑制剂SQ-22536引起THP-1巨噬细胞源性泡沫细胞总胆固醇、游离胆固醇与胆固醇酯增加. 载脂蛋白A-Ⅰ引起THP-1巨噬细胞源性泡沫细胞ABCA1蛋白质水平和细胞内胆固醇流出增加. FRK引起THP-1巨噬细胞源性泡沫细胞ABCA1蛋白质水平和细胞内胆固醇流出呈时间和浓度依赖性增加. SQ-22536引起THP-1巨噬细胞源性泡沫细胞ABCA1蛋白质水平和细胞内胆固醇流出减少. 结果提示,载脂蛋白A-Ⅰ可提高THP-1巨噬细胞源性泡沫细胞ABCA1蛋白质水平,增加细胞内胆固醇流出, 降低细胞内胆固醇聚积. 其机制可能是通过PKA信号途经使细胞ABCA1蛋白质水平增加.

    Abstract:

    ATP-binding cassette transporter A1 (ABCA1) plays a crucial role in apoA-Ⅰ binding activity and promotes cellular cholesterol efflux. In order to investigate the effect of interaction between apoA-Ⅰ and ABCA1 on atherosclerosis, and to discover the mechanism of interaction between apoA-Ⅰ and ABCA1, THP-1 cells were induced to become the macrophages by the phorbol. Then THP-1 macrophages were induced to the foam cells by ox-LDL. Treatment of THP-1 macrophage derived foam cells with apoA-Ⅰ, forskolin (FRK, an adenyl cyclase activator), and SQ-22536 (an adenyl cyclase inhibitor) for long periods of time (24 h). In addition, THP-1 macrophage derived foam cells were treated with increasing amounts of FRK (0, 10, 20, 40 and 80 μmol/L) and treated with FRK for increasing time(0, 6, 12 and 24 h). Cholesterol efflux, ABCA1 mRNA and protein level were determined by FJ-2107P type liquid scintillator, reverse transcriptase-polymerase chain reaction and Western blotting, respectively. Cellular lipid accumulation was determined by Oil Red O staining and high performance liquid chromatography analysis. The cholesterol efflux of control group, apoA-Ⅰ group, FRK group, apoA-Ⅰ+ FRK group and apoA-Ⅰ+SQ-22536 group was (8.64 ± 0.83)%, (9.8 ± 0.93)%, (10.15 ± 0.98)%, (11.72 ± 1.1)%, and (6.77 ± 0.7)%, respectively. ApoA-Ⅰ resulted in a 26.7% increase in protein expression of ABCA1, and a 14.0% increase in cholesterol efflux from THP-1 macrophage derived foam cells (P < 0.05). The similar results have been observed in foam cells treated with FRK. ApoA-Ⅰ, in combination with FRK, contributed to a much larger increase in protein expression of ABCA1 (80%, P < 0.05) and cholesterol efflux (36%, P < 0.05) from THP-1 macrophage derived foam cells. In converse, treatment THP-1 macrophage derived foam cells with apoA-Ⅰ and SQ-22536 markedly down-regulated protein expression of ABCA1 (26.7%, P < 0.05) and decreased cholesterol efflux (22.1%, P < 0.05). However, apoA-Ⅰ, FRK and SQ-22536 alone or combination did not influence mRNA expression of ABCA1. These findings suggest that apoA-Ⅰ may stabilize ABCA1 protein, and then increase cellular cholesterol efflux through PKA signaling.

    参考文献
    相似文献
    引证文献
引用本文

杨峻浩,代小艳,欧 翔,郝新瑞,曹冬黎,姜志胜,刘录山,王佐,易光辉,危当恒,王贵学,唐朝克.载脂蛋白A-Ⅰ通过PKA信号途径影响ABCA1的表达与功能[J].生物化学与生物物理进展,2007,34(6):611-619

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:2006-12-05
  • 最后修改日期:2007-02-06
  • 接受日期:
  • 在线发布日期: 2007-05-22
  • 出版日期: 2007-06-20