Stimulated human neutrophils limit iron-catalyzed hydroxyl radical formation as detected by spin-trapping techniques.
Stimulated human neutrophils limit iron-catalyzed hydroxyl radical formation as detected by spin-trapping techniques.
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DOI：
10.1016/s0021-9258(19)75994-8
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发表时间：
1986-12
期刊：
影响因子：
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通讯作者：
B. Britigan;G. Rosen;B. Thompson;Y. Chai;M. Cohen
中科院分区：
文献类型：
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作者：
B. Britigan;G. Rosen;B. Thompson;Y. Chai;M. Cohen
在自旋陷阱5,5-二甲基-1-吡啶氧化物（DMPO）、二甲亚砜和二乙烯三胺五乙酸（DETAPAC）存在的情况下，肉豆酸酯佛酚（PMA）刺激的中性粒细胞不能产生羟基自由基。OH)，被检测为DMPO（2,2,5-三甲基-1-吡啶基氧基，DMPO- ch3）的甲基自旋捕获加合物，除非还添加铁盐（Fe3+） (Britigan, b.e., Rosen, g.m., Chai， Y., and Cohen, m.s. (1986) J. Biol。化学，261,4426 -4431)。即便如此，……尽管持续产生超氧化物（O2-），但OH的形成减弱。相反，铁盐中添加一种次黄嘌呤/黄嘌呤氧化酶O2-。包含DETAPAC的发电系统连续生产。OH，表明中性粒细胞限制了自由基的形成。为了验证这一假设，在Fe3+、DETAPAC、二甲亚砜和DMPO存在的情况下，用PMA刺激中性粒细胞细胞质（大部分没有颗粒但能够产生O2-）。这导致了DMPO-CH3的持续产生。在二甲亚砜存在下，HL-60（早幼粒细胞）细胞分化为形态和O2-相似的细胞。产生中性粒细胞的能力。然而，它们的颗粒缺乏铁结合蛋白乳铁蛋白（LF）。添加铁盐的HL-60细胞受PMA刺激后，EPR谱与细胞质相似。pma诱导的中性粒细胞脱粒后获得的上清（在细胞外释放LF）抑制了次黄嘌呤/黄嘌呤氧化酶/Fe3+/DETAPAC系统形成DMPO-CH3。抗lf抗体，而非抗转铁蛋白抗体，可阻止受刺激的中性粒细胞上清对次黄嘌呤/黄嘌呤氧化酶/Fe3+/ detapac介导的抑制。哦的形成。同样，在Fe3+、DETAPAC和抗lf抗体（但不包括抗转铁蛋白抗体）存在的情况下，PMA刺激的中性粒细胞显示出持续形成。oh。LF中性粒细胞脱粒限制Fe3+催化。OH的形成在体内可以保护组织免受。OH-mediated受伤。
Neutrophils stimulated with phorbol myristate acetate (PMA) in the presence of the spin trap 5,5-dimethyl-1-pyrroline 1-oxide (DMPO), dimethyl sulfoxide, and diethylenetriaminepentaacetic acid (DETAPAC) fail to generate hydroxyl radical (.OH), detected as the methyl spin-trapped adduct of DMPO (2,2,5-trimethyl-1-pyrrolidinyloxyl, DMPO-CH3), unless ferric salts (Fe3+) are also added (Britigan, B. E., Rosen, G. M., Chai, Y., and Cohen, M. S. (1986) J. Biol. Chem. 261, 4426-4431). Even then, .OH formation wanes in spite of ongoing superoxide (O2-.) production. In contrast, ferric salt supplementation of a hypoxanthine/xanthine oxidase O2-. generating system containing DETAPAC produces continual .OH, suggesting that neutrophils limit the formation of this free radical. To evaluate this hypothesis, neutrophil cytoplasts (largely devoid of granules but able to generate O2-.) were stimulated with PMA in the presence of Fe3+, DETAPAC, dimethyl sulfoxide, and DMPO. This resulted in continual production of DMPO-CH3. In the presence of dimethyl sulfoxide, HL-60 (promyelocytic) cells differentiate into cells similar in morphology and O2-. generating capacity to neutrophils. However, their granules lack the iron-binding protein lactoferrin (LF). Ferric salt supplementation of HL-60 cells stimulated with PMA yielded an EPR spectrum similar to cytoplasts. Supernatant obtained following PMA-induced neutrophil degranulation (which releases LF extracellularly) suppressed DMPO-CH3 formation by the hypoxanthine/xanthine oxidase/Fe3+/DETAPAC system. Anti-LF antibody, but not anti-transferrin antibody, prevented stimulated neutrophil supernatant inhibition of hypoxanthine/xanthine oxidase/Fe3+/DETAPAC-mediated .OH formation. Similarly, neutrophils stimulated with PMA in the presence of Fe3+, DETAPAC, and anti-LF antibody (but not anti-transferrin antibody) demonstrated continual formation of .OH. Neutrophil degranulation of LF limits Fe3+-catalyzed .OH formation which in vivo could protect tissue from possible .OH-mediated injury.