Lacy F, Gough D A, Schmid-Schönbein G W
Department of Bioengineering, Institute for Biomedical Engineering, University of California at San Diego, La Jolla 92093-0412, USA.
Free Radic Biol Med. 1998 Oct;25(6):720-7. doi: 10.1016/s0891-5849(98)00154-3.
Increased production of oxygen free radicals may play a role in many diseases such as hypertension. As evidence indicates that xanthine oxidase may be involved in creating these reactive oxygen species, experiments were performed to additionally characterize hydrogen peroxide (H2O2) production in xanthine oxidase catalyzed reactions. In vitro measurements of hydrogen peroxide production from the xanthine/xanthine oxidase reaction were performed in buffered saline using an electrochemical technique, and the effect of allopurinol on inhibition of xanthine oxidase was determined. Experiments were also performed in blood plasma to characterize endogenous hydrogen peroxide producing capability and xanthine oxidase activity. In the presence of sodium azide, an inhibitor of catalase, peroxide production was measured in plasma after adding xanthine or xanthine oxidase and the results were similar to those obtained in buffered saline. When only sodium azide was added to plasma, hydrogen peroxide was produced at a level of 36.1 +/- 7.6 microM (n = 5). From these measurements, endogenous xanthine oxidase activity was estimated to be 6.5 +/- 0.3 mU/ml (n = 5). These results suggests that sufficient substrate exists in plasma to produce micromolar levels of hydrogen peroxide and xanthine oxidase may catalyze these reactions.
氧自由基生成增加可能在许多疾病如高血压中起作用。有证据表明黄嘌呤氧化酶可能参与产生这些活性氧,因此进行了实验以进一步表征黄嘌呤氧化酶催化反应中过氧化氢(H2O2)的生成情况。使用电化学技术在缓冲盐溶液中对黄嘌呤/黄嘌呤氧化酶反应产生的过氧化氢进行体外测量,并测定了别嘌呤醇对黄嘌呤氧化酶的抑制作用。还在血浆中进行了实验以表征内源性过氧化氢生成能力和黄嘌呤氧化酶活性。在存在过氧化氢酶抑制剂叠氮化钠的情况下,加入黄嘌呤或黄嘌呤氧化酶后测量血浆中的过氧化物生成,结果与在缓冲盐溶液中获得的结果相似。当仅向血浆中加入叠氮化钠时,过氧化氢的生成水平为36.1±7.6微摩尔(n = 5)。根据这些测量结果,内源性黄嘌呤氧化酶活性估计为6.5±0.3毫单位/毫升(n = 5)。这些结果表明血浆中存在足够的底物以产生微摩尔水平的过氧化氢,并且黄嘌呤氧化酶可能催化这些反应。