SCATCHARD ANALYSIS OF METHANE SULFINIC ACID PRODUCTION FROM DIMETHYL-SULFOXIDE - A METHOD TO QUANTIFY HYDROXYL RADICAL FORMATION IN PHYSIOLOGIC SYSTEMS
SCATCHARD ANALYSIS OF METHANE SULFINIC ACID PRODUCTION FROM DIMETHYL-SULFOXIDE - A METHOD TO QUANTIFY HYDROXYL RADICAL FORMATION IN PHYSIOLOGIC SYSTEMS
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DOI：
10.1016/0891-5849(89)90042-7
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发表时间：
1989-01-01
影响因子：
7.4
通讯作者：
GRIFFIN, DW
中科院分区：
文献类型：
--
作者：
BABBS, CF;GRIFFIN, DW
确认自由基发病机制的一个主要障碍是缺乏直接的化学证据，证明以氧为中心的自由基实际上在活组织中产生的数量足以造成严重损害。进行该研究以验证使用二甲基亚砜（DMSO）作为用于产生羟基自由基（HO.在生理条件下。HO.cntdot.与DMSO反应产生甲烷亚磺酸（MSA）作为主要产物，其可以通过简单的比色测定来检测。开发一种估算总HO的方法。本论文以黄嘌呤氧化酶为超氧化物源，研究了两个模型体系：超氧化物驱动的体外芬顿反应和芬顿化学的计算机模型。在体外和计算机模型中测量的MSA产生是DMSO和HO的竞争清除剂的浓度的可预测函数。根据竞争动力学原理，实验结果和模型计算表明，Scatchard分析可以用来推断总HO。尽管存在除DMSO以外的清除剂，如甘露醇，但仍然可以产生。因此，从DMSO产生的甲烷亚磺酸有望作为HO的容易测量的标志物。在用DMSO预处理的生物系统中的总HO形成，并且用不同DMSO浓度的重复实验的Scatchard分析可以产生总HO的估计。一代
A major impediment to the confirmation of free radical mechanisms in pathogenesis is a lack of direct, chemical evidence that oxygen centered free radicals actually arise in living tissues in quantities sufficient to cause serious damage. This investigation was conducted to validate the use of dimethyl sulfoxide (DMSO) as a quantitative molecular probe for the generation of hydroxyl radicals (HO.cntdot.) under physiologic conditions. Reaction of HO.cntdot. with DMSO produces methane sulfinic acid (MSA) as a primary product, which can be detected by a simple colorimetric assay. To develop a method for estimating total HO.cntdot. production, we studied two model systems: the superoxide driven Fenton reaction in vitro, usign xanthine oxidase as the source of superoxide, and a computer model of Fenton chemistry. Measured MSA production both in vitro and in the computer model was a predictable function of the concentrations of DMSO and competing scavengers of HO.cntdot. according to the principle of competition kinetics. Both experimental results and model calculations showed that Scatchard analysis may be used to infer total HO.cntdot. generation, despite the presence of scavengers other than DMSO, such as mannitol. Thus, methane sulfinic acid production from DMSO holds promise as an easily measured marker for HO.cntdot. formation in biologic systems pretreated with DMSO, and Scatchard analysis of repeated experiments with varying DMSO concentrations can yield an estimate of total HO.cntdot. generation.