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[人白细胞髓过氧化物酶制剂杀念珠菌活性的体外评估]

[In vitro assessment of candidacidal activity of human leukocyte myeloperoxidase preparation].

作者信息

Li C L, Wang B Y, Jiang Z Q, Li X A

出版信息

Hua Xi Yi Ke Da Xue Xue Bao. 1989 Mar;20(1):39-41.

PMID:2551812
Abstract

Myeloperoxidase (MPO), present in the azurophilic granules of human polymorphonuclear neutrophils, is important in the oxygen-dependent microbicidal activity of neutrophils. The purpose of our study was to investigate the therapeutic potency of the MPO preparations. This paper is to present our primary work on MPO isolation and its microbicidal activity assay. White blood cells, isolated freshly from normal donors, were lysed with cetyltrimehylammonium bromide to liberate myeloperoxidase. The enzyme preparation was partially purified by 50% (NH4)2SO4 precipitation followed by 65% (NH4)2SO4 precipitation. A hundred million leukocytes yielded 1.03 mg protein of the MPO preparation with the RZ of 0.31. The specific activity of the MPO preparation was about 29.25 u/mg. When 0.672 units of the MPO preparation were incubated with about 10(7) clinical isolates of Candida albicans in the presence of 0.2 mmol/L H2O2 and 0.14 mol/L NaCl. It was detected that 95.58 +/- 0.64% of the organisms were killed in the methylene blue staining system.

摘要

髓过氧化物酶(MPO)存在于人类多形核中性粒细胞的嗜天青颗粒中,在中性粒细胞的氧依赖性杀菌活性中起重要作用。我们研究的目的是调查MPO制剂的治疗效力。本文将介绍我们关于MPO分离及其杀菌活性测定的初步工作。从正常供体新鲜分离的白细胞用十六烷基三甲基溴化铵裂解以释放髓过氧化物酶。酶制剂先通过50%硫酸铵沉淀,然后通过65%硫酸铵沉淀进行部分纯化。一亿个白细胞产生了1.03毫克蛋白质的MPO制剂,其RZ值为0.31。MPO制剂的比活性约为29.25单位/毫克。当0.672单位的MPO制剂在0.2毫摩尔/升过氧化氢和0.14摩尔/升氯化钠存在的情况下与约10⁷株白色念珠菌临床分离株一起孵育时,在亚甲蓝染色系统中检测到95.58±0.64%的微生物被杀死。

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