Badwey J A, Curnutte J T, Robinson J M, Lazdins J K, Briggs R T, Karnovsky M J, Karnovsky M L
J Cell Physiol. 1980 Dec;105(3):541-5. doi: 10.1002/jcp.1041050319.
We have compared the subcellular sites of H2O2 and presumably also superoxide-(O2-) production, and certain aspects of metabolic responses (O2 consumption, O2- production) of stimulated neutrophils from human blood and those elicited into guinea pig peritonea. Stimulation was accomplished with either opsonized zymosan or phorbol-12-myristate-13-acetate (PMA). Striking quantitative differences were observed between these cell types with regard to the increased respiration and O2- production observed during stimulation. These differences were most apparent when opsonized zymosan served as the stimulating agent. They were minimized when the soluble stimulating agent, PMA, was used. With either stimulus, the subcellular sites of H2O2 production were the same for both types of neutrophils, i.e., the plasmalemma and phagosomal membranes. No H2O2 production could be detected cytochemically in the absence of stimulation. Treatment of both unstimulated human blood and elicited guinea pig peritoneal neutrophils with the nonpenetrating, covalently linking reagent, p-diazobenzenesulfonic acid, failed to diminish O2- production upon subsequent stimulation, in contrast to a previous report. These data are discussed in terms of the possible cytological arrangements of the respiratory enzyme(s), and the different modes of stimulation of neutrophil metabolism by various agents. Ancillary data on elicited mouse peritoneal neutrophils are presented.
我们比较了过氧化氢以及可能还有超氧阴离子(O₂⁻)产生的亚细胞位点,以及来自人血的受刺激中性粒细胞和豚鼠腹膜中诱发的中性粒细胞的某些代谢反应方面(氧气消耗、O₂⁻产生)。刺激是用调理酵母聚糖或佛波醇-12-肉豆蔻酸酯-13-乙酸酯(PMA)完成的。在刺激过程中观察到的呼吸增加和O₂⁻产生方面,这些细胞类型之间存在显著的数量差异。当调理酵母聚糖作为刺激剂时,这些差异最为明显。当使用可溶性刺激剂PMA时,差异最小化。无论使用哪种刺激,两种类型的中性粒细胞中过氧化氢产生的亚细胞位点都是相同的,即质膜和吞噬体膜。在没有刺激的情况下,细胞化学方法检测不到过氧化氢的产生。与之前的一份报告相反,用非穿透性的、共价连接试剂对二氮苯磺酸处理未受刺激的人血中性粒细胞和诱发的豚鼠腹膜中性粒细胞,在随后的刺激时未能减少O₂⁻的产生。根据呼吸酶可能的细胞学排列以及各种试剂刺激中性粒细胞代谢的不同方式对这些数据进行了讨论。还给出了诱发的小鼠腹膜中性粒细胞的辅助数据。