摘要
本文采用靛蓝胭脂红褪色反应检测抗体是否能催化经佛波酯(PMA)激活的THP-1单核细胞产生臭氧(O_3),并借助荧光分光光度法检测细胞内胆固醇的积聚,以及高效液相色谱法(HPLC)分析细胞内胆固醇的臭氧氧化产物5,6-secosterol生成的情况,探讨抗体催化水的氧化作用对动脉粥样病变形成潜在的影响。结果显示,THP-1单核细胞经人IgG、PMA共同孵育后,显著产生了O_3的化学信号,对靛蓝胭脂红具有明显的褪色作用,过氧化氢酶能加强这种褪色反应,而乙烯苯甲酸具有显著的抑制作用；THP-1单核细胞经人IgG、PMA及低密度脂蛋白(LDL)共同孵育后,其细胞内积聚的总胆固醇(TC)、游离胆固醇(FC)、胆固醇酯(CE)及胆固醇酯占总胆固醇的比值均明显增多,并显著产生了胆固醇的臭氧氧化产物5,6-secosterol,乙烯苯甲酸对它们均有抑制效应。以上结果表明：激活的THP-1单核细胞在抗体的催化下具有产生臭氧的能力,可能是动脉粥样硬化形成过程中一种新的重要机制。
In the present study, we measured the antibody-catalyzed 03 formation from THP-1 monocytes activated by phorbol myristate acetate （PMA） by indigo carmine bleaching reaction test,and the accumulation of cholesterol in THP-1 monocytes by fluorescence spectrophotometric method,and analyzed the cholesterol ozonation product 5,6-secosterol by high-performance liquid chromatography （HPLC） ,to explore the potential effect of antibody-catalyzed water oxidation on pathogenesis of atherosclerosis. It was showed that THP-1 monocytes incubated with human IgG and PMA evidently produced an oxidant with the chemical signature of O3 which could bleach indigo carmine, and be intensified or inhibited respectively by catalase and vinylbenzoic acid. In the THP-1 monocytes incubated with human IgG,PMA and LDL,the intracellular accumulated total cholesterol （TC）,free cholesterol （FC）,cholesteryl ester （CE） and the CE/TC increased evidently, and the cholesterol ozonation product 5,6-secosterol was also produced markly,all of that were inhibited by vinylbenzoic acid. These results demonstrated that the activated THP-1 monocytes possess the ability to produce O3 through antibody-catalyzed water-oxidation pathway,which could be a new mechanism concerned with atheriosclerosis.
出处
《分子细胞生物学报》
SCIE
CAS
CSCD
北大核心
2006年第6期509-515,共7页
Journal of Molecular Cell Biology
基金
教育部博士点基金项目(20050610050)
