Arnhold J, Hammerschmidt S, Arnold K
Institute of Biophysics, School of Medicine, University of Leipzig, F.R.G.
Biochim Biophys Acta. 1991 Sep 23;1097(2):145-51. doi: 10.1016/0925-4439(91)90099-u.
Hypochlorous acid HOCl/OCl- and other oxidants derived from stimulated polymorphonuclear leukocytes are involved in tissue damage during a number of pathological processes. In order to obtain more detailed information on possible reactions of HOCl/OCl- the effects of both NaOCl and PMN-derived hypochlorous acid on functional groups of amino acid solutions and human plasma are studied. In valine and lysine solutions NaOCl diminishes the number of amino groups in a molar ratio of 1:1 between NaOCl and amino groups. In cysteine and methionine samples the decrease of amino groups starts only after all sulfhydryl or thioether groups are oxidized by NaOCl. If freshly prepared human plasma is treated with increasing amounts of NaOCl all plasma SH groups are oxidized first, then probably the thioether groups and only after this the amino groups are affected. Furthermore, it was found, that the reactivity of luminol against NaOCl is similar to that of amino groups. Increasing amounts of SH groups of components of human plasma are oxidized by incubation with PMA-stimulated polymorphonuclear leukocytes dependent on the incubation time. Plasma amino groups are not affected under the same experimental conditions. The addition of plasma to FMLP-stimulated PMN in the presence of luminol decreases that part of chemiluminescence caused by extracellularly generated hypochlorous acid. Plasma samples pretreated with NaOCl cause a lower inhibition of light generation in FMLP-stimulated PMN only when more than 4.10(-8) mol NaOCl per mg protein are used to pretreat plasma. It is assumed that in the development of tissue injuries caused by infiltrated PMN the following sequence of damage occurs in accessible tissue regions. First, the sulfhydryl groups are oxidized, then the thioether groups, and only after this amino and other target groups are affected.
次氯酸HOCl/OCl⁻以及来自受刺激多形核白细胞产生的其他氧化剂,在许多病理过程中参与组织损伤。为了获得关于HOCl/OCl⁻可能反应的更详细信息,研究了次氯酸钠(NaOCl)和多形核白细胞衍生的次氯酸对氨基酸溶液和人血浆官能团的影响。在缬氨酸和赖氨酸溶液中,NaOCl以NaOCl与氨基1:1的摩尔比减少氨基数量。在半胱氨酸和蛋氨酸样品中,只有当所有巯基或硫醚基团被NaOCl氧化后,氨基数量才开始减少。如果用越来越多的NaOCl处理新鲜制备的人血浆,所有血浆中的巯基首先被氧化,然后可能是硫醚基团,只有在此之后氨基才会受到影响。此外,还发现鲁米诺对NaOCl的反应性与氨基相似。通过与佛波酯(PMA)刺激的多形核白细胞孵育,人血浆成分中越来越多的巯基会被氧化,这取决于孵育时间。在相同实验条件下,血浆氨基不受影响。在鲁米诺存在的情况下,向受N-甲酰甲硫氨酸-亮氨酸-苯丙氨酸(FMLP)刺激的多形核白细胞中添加血浆会降低由细胞外产生的次氯酸引起的那部分化学发光。只有当每毫克蛋白质使用超过4×10⁻⁸摩尔NaOCl预处理血浆时,用NaOCl预处理的血浆样品对FMLP刺激的多形核白细胞中光产生的抑制作用才会降低。据推测,在由浸润的多形核白细胞引起的组织损伤发展过程中,在可及的组织区域会发生以下损伤顺序。首先,巯基被氧化,然后是硫醚基团,只有在此之后氨基和其他靶基团才会受到影响。