NF-κB在AngⅡ介导THP-1巨噬细胞源性泡沫细胞胆固醇流出及AB-CA1表达调控过程中的作用研究  

Effect of nuclear factor-κB on AngⅡ induced ABCA1 expression and cellular cholesterol efflux in THP-1 derived-macrophage foam cells

作　　者：岳兵[1] 陈志坚[1] 王彦富[1] 彭红玉[1] 梅春丽[1] 肖华[1] 祝聪聪[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院心血管病研究所,武汉430022

出　　处：《中国免疫学杂志》2008年第5期398-402,共5页Chinese Journal of Immunology

摘　　要：目的:探讨核因子NF-κB活化对AngⅡ诱导的THP-1巨噬细胞源性泡沫细胞ABCA1基因表达、胆固醇含量的影响。方法:体外培养THP-1细胞以构建泡沫细胞模型,以AngⅡ和NF-κB活化抑制剂TPCK孵育细胞;以RT-PCR法、Western blot法测定THP-1源泡沫细胞ABCA1 mRNA和蛋白表达状态;采用酶法,通过荧光分光光度计检测细胞内胆固醇含量,应用液体闪烁技术仪检测胆固醇流出的变化;借助Sandwich ELISA法测定核因子NF-κB活化/核易位情况。结果:AngⅡ可即刻激活NF-κB表达活性,并导致干预24小时后泡沫细胞ABCA1 mRNA和蛋白表达下调;若预先用TPCK孵育,则TPCK可抑制AngⅡ对NF-κB的激活,且24小时后泡沫细胞ABCA1 mRNA和蛋白表达下调幅度减小。结论:AngⅡ能引起THP-1源性泡沫细胞胆固醇含量显著增高(P<0·05)、ABCA1表达显著减少(P<0·05)。机制可能与AngⅡ即刻激活NF-κB信号途径,早期活化的NF-κB可阻遏THP-1源泡沫细胞ABCA1基因和蛋白的表达,继而影响泡沫细胞内胆固醇的流出有关。Objective： To investigate whether nuclear factor（NF-κB） activation participate in the regulation and control of ABCA1 gene expression and cellular cholesterol efflux in THP-1 derived-macrophage foam cells. Methods： First, THP-1 cells were be transformed into foam cells.Then,foam cells were treated with AngⅡ （10 ng/ml） ,or with TPCK（10 μmol/L） at 60 rain before Angn stimulation or at different time paints after AngⅡ stimulation. Gene expression of ABCA1 was analyzed using RT-PCR and its protein was detected by Western blot. Sandwich ELISA method was employed to evaluate the nuclear translocation of NF-κB subunit p65. Results： Ang Ⅱ could down regulate both ABCA1 mRNA expression and ABCA1 protein level. Stimulation of foam cells by AngⅡ led to a Iapid increase of NF-κB 1365 nuclear translocation and the peak of NF-κB 1365 was 30-60 min. Addition of TPCK to the cells at 60 min before AngⅡ attenuated the increase of nuclear factor（NF-κB） translocated into the nucleus and the decrease of ABCA1 gene expression induced by AngⅡ . Condusion：AngⅡ could swiftly result in NF-κB activation.Activation of NF-κB signal transduction may finally inhibit ABCA1 gene expression, which may play a critical role in cellular cholesterol efflux.

关 键 词：TFIP-1 核因子ΚB ATP结合盒转运子A1 对甲苯磺酰-L-苯丙氨酸甲基甲酮 细胞胆固醇外流 

分 类 号：R392.12[医药卫生—免疫学]