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目的:探讨核因子-κB(NF-κB)对血管紧张素Ⅱ(AngⅡ)诱导THP-1源性泡沫细胞ATP结合盒转运子A1(ABCA1)表达的影响及在胆固醇流出中的作用。方法:THP-1源性泡沫细胞分别给予10-5mol/LAngⅡ(AngⅡ组)、10μmol/LNF-κB特异性抑制剂TPCK预孵(TPCK组)。采用夹心酶联免疫分析法检测泡沫细胞内NF-κB的活化程度。通过透射电镜和荧光分光光度计、酶化学法,检测泡沫细胞内胆固醇含量的变化。利用闪烁计数法测量泡沫细胞内胆固醇流出率。用逆转录-聚合酶链反应法和免疫印迹法半定量分析泡沫细胞ABCA1的表达。结果:TPCK组NF-κB活化核易位则无明显高峰,维持在较低的水平。TPCK组泡沫细胞内胆固醇含量较AngⅡ组降低24.1%(P<0.05),胆固醇流出率较AngⅡ组显著升高41.1%(P<0.05),ABCA1mR-NA和蛋白表达较AngⅡ组升高30%和19%(P<0.05)。结论:AngⅡ可经NF-κB介导下调THP-1源性泡沫细胞ABCA1的表达,致泡沫细胞内胆固醇流出减少,增加泡沫细胞内胆固醇含量,加速动脉粥样硬化。
Objective: To investigate the effect of nuclear factor-κB (NF-κB) on the expression of ATP-binding cassette transporter A1 (ABCA1) in THP-1-derived foam cells induced by Angiotensin Ⅱ (Angiotensin Ⅱ) and its role in cholesterol efflux. Methods: THP-1-derived foam cells were pretreated with TPCK (10-5mol / LAngⅡ) and TPCK (10μmol / LNF-κB specific inhibitor) respectively. Sandwich enzyme-linked immunosorbent assay was used to detect the activation of NF-κB in foam cells. The changes of cholesterol content in foam cells were detected by transmission electron microscopy, fluorescence spectrophotometer and enzyme chemistry. The scintillation counting method was used to measure the rate of cholesterol efflux in foam cells. Semi - quantitative analysis of ABCA1 expression in foam cells by reverse transcription - polymerase chain reaction and Western blotting. Results: There was no obvious peak of nuclear translocation of NF-κB in TPCK group, and remained at a low level. Compared with AngⅡ group, the cholesterol efflux in TPCK group decreased by 24.1% (P <0.05) and that in AngⅡ group increased by 41.1% (P <0.05), and ABCA1mR-NA and protein expression increased by 30% 19% (P <0.05). CONCLUSION: AngⅡ can down-regulate the expression of ABCA1 in THP-1-derived foam cells via NF-κB, resulting in the decrease of intracellular cholesterol efflux in foam cells, increasing the content of cholesterol in foam cells and accelerating atherosclerosis.