Dallegri F, Goretti R, Ballestrero A, Ottonello L, Patrone F
First Medical Clinic, University of Genoa Medical School, Italy.
J Lab Clin Med. 1988 Dec;112(6):765-72.
Human neutrophils, incubated with phorbol myristate acetate (PMA), caused a rapid and substantial adenosine triphosphate (ATP) depletion in lymphoblastoid Daudi cells without producing lysis. Catalase (which destroys hydrogen peroxide), taurine and methionine (which scavenge hypochlorous acid), and chloride omission from the medium prevented the ATP fall. An ATP depletion comparable to that induced by neutrophils was observed by replacing neutrophils with an appropriate myeloperoxidase-H2O2-Cl- enzymatic system. Together, these data suggest that the neutrophil ATP depleting activity involves the myeloperoxidase-catalyzed transformation of H2O2 into HOCl. Moreover, the free H2O2 remaining in the neutrophil extracellular environment is ineffective. In fact, a comparable amount of enzymatically generated H2O2 did not cause Daudi cell ATP loss. A direct role for H2O2 in the neutrophil-induced Daudi cell ATP depletion was observed only under artificial conditions, that is, in the presence of the heme enzyme inhibitor azide, which prevented the HOCl production but dramatically augmented the extracellular H2O2 level. Similar levels of ATP depletion in Daudi cells were induced by amounts of reagent HOCl comparable to those generated by neutrophils. As the generated HOCl can rapidly react with a variety of neutrophil-derived nitrogenous compounds (primarily ammonia and taurine) to yield chloramines, these chlorinated oxidants might contribute to the neutrophil-mediated ATP depletion. Nevertheless, the main and well-characterized chloramines (ammonia-derived monochloramine, NH2Cl, and taurine monochloramine, TauNHCl) were devoid of ATP-depleting capacity. Thus, the results suggest that the neutrophil-induced ATP depletion in Daudi cells is HOCl-dependent, is not mediated by NH2Cl or TauNHCl, and could be promoted either by HOCl directly or by an unknown derivative oxidant.(ABSTRACT TRUNCATED AT 250 WORDS)
人中性粒细胞与佛波醇肉豆蔻酸酯乙酸酯（PMA）孵育后，可使淋巴母细胞样Daudi细胞中的三磷酸腺苷（ATP）迅速大量消耗，且不引起细胞裂解。过氧化氢酶（可破坏过氧化氢）、牛磺酸和蛋氨酸（可清除次氯酸）以及培养基中去除氯离子可防止ATP水平下降。用适当的髓过氧化物酶-H2O2-Cl-酶系统替代中性粒细胞，可观察到与中性粒细胞诱导的ATP消耗相当的情况。综合这些数据表明，中性粒细胞的ATP消耗活性涉及髓过氧化物酶催化H2O2转化为HOCl。此外，中性粒细胞细胞外环境中残留的游离H2O2无效。事实上，等量酶促产生的H2O2不会导致Daudi细胞ATP损失。仅在人工条件下，即在存在血红素酶抑制剂叠氮化物的情况下，观察到H2O2在中性粒细胞诱导的Daudi细胞ATP消耗中起直接作用，叠氮化物可阻止HOCl产生，但显著提高细胞外H2O2水平。与中性粒细胞产生的量相当的试剂HOCl可诱导Daudi细胞中类似水平的ATP消耗。由于产生的HOCl可迅速与多种中性粒细胞衍生的含氮化合物（主要是氨和牛磺酸）反应生成氯胺，这些氯化氧化剂可能有助于中性粒细胞介导的ATP消耗。然而，主要且特征明确的氯胺（氨衍生的一氯胺，NH2Cl，和牛磺酸一氯胺，TauNHCl）没有ATP消耗能力。因此，结果表明中性粒细胞诱导的Daudi细胞ATP消耗依赖于HOCl，不是由NH2Cl或TauNHCl介导的，并且可能由HOCl直接或由未知的衍生氧化剂促进。（摘要截短至250字）
