抗体催化的臭氧生成及其在动脉粥样硬化发病中的潜在作用

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。

ANTIBODY-INDUCED OZONE PRODUCTION AND ITS POTENTIAL EFFECTS ON PATHOGENESIS OF ATHEROSCLEROSIS

In the present study,we measured the antibody-catalyzed O_3 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 O_3 which could bleach in- digo 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 03 through antibody-catalyzed water-oxidation pathway,which could be a new mechanism concerned with atheriosclerosis.