用化学发光法研究NADPH氧化酶产生O_2~+的活性

本文用化学发光法研究了从促癌剂PMA刺激的人血多形核白细胞中提取的NADPH氧化酶在全溶无细胞体系中产生O_2~+的活性.给出了其发光值随时间的变化曲线;在相同条件下,其活性比从未刺激的多形核白细胞中提取的NADPH氧化酶约大4.5倍,与细胞色素C还原—SOD抑制法所得结果相一致.

SIUDIES ON THE SUPEROXIDE-GENERATING ACTIVITY OF NADPH OXIDASE BY CHEM1LUMINESCENCE

The respiratory burst oxidase(or NADPH oxidase)is a membrane-bound enzyme which is responsible for the production of superoxide anions by phagocytes. The activity of the NADPH oxidase from PMA-stimulated and control human PMN in a fully soluble cell-free system was studied by luminol-dependent chemilumine-scence(CL). The CL value increased with the NADPH concentration, and increased linearly with the concentration of the enzyme. The activity of the NADPH oxidase of PMA-stimulated PMN was much higher than that of the control PMN at the same concentration of NADPH. The time-course of CL value was presented. In addition, the rate of superoxide anion production determined by the superoxide-dismutasc (SOD) sensitive ferricytochrome-C reduction was studied in comparison. We presume that the process of PMA stimulation is probably an assembly of the components of the NADPH oxidase.