Regulation of apoptosis and priming of neutrophil oxidative burst by diisopropyl fluorophosphate.
Regulation of apoptosis and priming of neutrophil oxidative burst by diisopropyl fluorophosphate.
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DOI：
10.1186/1476-9255-7-32
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发表时间：
2010-07-07
期刊：
影响因子：
--
通讯作者：
Marshall JC
中科院分区：
文献类型：
--
作者：
Tsang JL;Parodo JC;Marshall JC
二异丙基氟磷酸（DFP）是一种丝氨酸蛋白酶抑制剂，在体外中性粒细胞研究中广泛用作内源性蛋白酶抑制剂。其对中性粒细胞功能的影响尚不清楚。我们试图确定DFP对人中性粒细胞凋亡和氧化破裂的生物学效应。我们从健康志愿者中分离出中性粒细胞，用DFP （2.5 mM）孵育，并评估中性粒细胞弹性酶（NE）活性、中性粒细胞脱粒、细胞凋亡（反映在次二倍体DNA形成和磷脂酰丝氨酸（PS）的外表面）、caspase -3和-8的加工和活性、氧化爆发活性和过氧化氢释放)。与其作为丝氨酸蛋白酶抑制剂的活性一致，DFP显著抑制NE活性，但不抑制亲氮颗粒的脱粒。DFP抑制组成性中性粒细胞凋亡，反映在DNA片段化，以及caspase -3和-8的加工和活性上。DFP还抑制中性粒细胞的氧化爆发活性和过氧化氢释放。然而，DFP以剂量依赖的方式增强了PS的外化。我们得出结论，DFP对中性粒细胞炎症功能有显著影响，这可能会混淆使用其抗蛋白酶活性的研究的解释。我们进一步得出结论，内源性蛋白酶在组成性中性粒细胞凋亡的生物学中发挥作用。
Diisopropyl fluorophosphate (DFP) is a serine protease inhibitor that is widely used as an inhibitor of endogenous proteases in in vitro neutrophil studies. Its effects on neutrophil function are unclear. We sought to determine the biological effects of DFP on human neutrophil apoptosis and oxidative burst. We isolated neutrophils from healthy volunteers, incubated them with DFP (2.5 mM), and evaluated neutrophil elastase (NE) activity, neutrophil degranulation, apoptosis as reflected in hypodiploid DNA formation and exteriorization of phosphatidylserine (PS), processing and activity of caspases-3 and -8, oxidative burst activity and hydrogen peroxide release. Consistent with its activity as a serine protease inhibitor, DFP significantly inhibited NE activity but not the degranulation of azurophilic granules. DFP inhibited constitutive neutrophil apoptosis as reflected in DNA fragmentation, and the processing and activity of caspases-3 and -8. DFP also inhibited priming of neutrophils for oxidative burst activity and hydrogen peroxide release. However, DFP enhanced the exteriorization of PS in a dose-dependent manner. We conclude that DFP exerts significant effects on neutrophil inflammatory function that may confound the interpretation of studies that use it for its antiprotease activity. We further conclude that endogenous proteases play a role in the biology of constitutive neutrophil apoptosis.
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影响因子：
1.6
作者：
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3.2
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通讯作者：
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