摘要
目的研究人红细胞保护蛋白 (HRPRP)在体外对羟基自由基 (·OH)的清除作用并探讨其抗氧化机制。方法可见光照射试管内N 羟基 2 硫代吡啶酮产生·OH ,用脱氧核糖作检测分子 ,分光光度法测定N 羟基 2 硫代吡啶酮随光照时间产生·OH的量。在反应体系中加入不同浓度的铜锌超氧化物歧化酶 (Cu ,Zn SOD)、HRPRP及二硫苏糖醇 (DTT)活化HRPRP ,观察它们对N 羟基 2 硫代吡啶酮产生的·OH的清除作用。结果Cu ,Zn SOD和HRPRP对·OH的清除率无显著性差异 (P >0 .0 5 ) ,DTT活化的HRPRP对·OH的清除率显著大于Cu ,Zn SOD(P <0 .0 1)及未活化HRPRP(P <0 .0 1)。结论证实HRPRP在巯基还原剂提供还原当量 ,暴露其自身活性位点时具有高效而特异的清除·OH的能力 ,提示HRPRP清除·OH可能是重要的抗氧化机制之一。
PurposeTo study the removal of hydroxyl radicals(·OH) by human red blood cell protect protein(HRPRP) in vitro.MethodsReactions that employed N-hydroxyl-2-thiopyridone (C 5H 5NOS) to generate ·OH were run in test tubes that contained 5mmol/L C 5H 5NOS and 10 mmol/L deoxyribose in 50 mmol/L buffer at pH 8.0. All tubes were capped and settled in front of the 200-W light bulb. During this reaction, one test tube was taken away from light every 1 h. The time course of ·OH release from C 5H 5NOS was analyzed. Then Cu,Zn-SOD, HRPRP and activated HRPRP were added to analyze their effects on ·OH.ResultsOur experiments showed that the absorbance of sample treated with activated HRPRP was remarkably less than that with Cu,Zn-SOD (P<0.01) or HRPRP (P<0.01). No significant discrepancy was found between Cu,Zn-SOD and HRPRP (P>0.01). The above results show that HRPRP scavenge ·OH when activated by dithiolthreitol.ConclusionRemoval of ·OH by HRPRP is involved in its antioxidant properties.
出处
《中国生化药物杂志》
CAS
CSCD
2004年第5期297-299,共3页
Chinese Journal of Biochemical Pharmaceutics
