Diphenyleneiodonium, an NAD(P)H Oxidase Inhibitor, also Potently Inhibits Mitochondrial Reactive Oxygen Species Production

Li, Yunbo; Trush, Michael A.

doi:10.1006/bbrc.1998.9729

Cited by 426 publications

(299 citation statements)

References 17 publications

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“…This oxidase activity has been assumed to be that of the NADPH oxidase, but mitochondrial function may now contribute to intracellular neutrophil oxidant production. Indeed, the most commonly used NADPH oxidase inhibitor is DPI (39), but this is also a known inhibitor of mitochondria (40). Third, it is known that neutrophils can generate reactive oxidant species intracellularly, which has again been assumed to be via internal NADPH oxidase activity (41)(42)(43)(44).…”

Section: Discussionmentioning

confidence: 99%

The Mitochondrial Network of Human Neutrophils: Role in Chemotaxis, Phagocytosis, Respiratory Burst Activation, and Commitment to Apoptosis

Fossati

Moulding

Spiller

et al. 2003

The Journal of Immunology

308

251

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It is commonly assumed that human neutrophils possess few, if any, functional mitochondria and that they do not depend on these organelles for cell function. We have used the fluorescent mitochondrial indicators, JC-1, MitoTracker Red, and dihydrorhodamine 123 to show that live neutrophils possess a complex mitochondrial network that extends through the cytoplasm. The membrane potential of these mitochondria was rapidly (within 2 min) disrupted by the addition of FCCP (IC50 = 20 nM), but not by the Fo-ATPase inhibitor, oligomycin (at up to 7 μg/ml). However, inhibition of mitochondrial function with both agents resulted in cell shape changes. Neither activation of the respiratory burst nor phagocytosis of either latex particles or serum-opsonized Staphylococcus aureus was affected by the addition of FCCP or oligomycin. However, FCCP inhibited chemotaxis at concentrations that paralleled disruption of mitochondrial membrane potential. Furthermore, prolonged (2-h) incubation with oligomycin resulted in an impaired ability to activate a respiratory burst and also inhibited chemotaxis. These observations indicate that intact mitochondrial function is required to sustain some neutrophil functions, but not for the rapid initiation of the respiratory burst or phagocytosis. Loss of mitochondrial membrane potential was a very early marker for commitment of neutrophils into apoptosis and preceded the appearance of phosphatidylserine on the cell surface. However, inhibition of mitochondrial function did not accelerate the rate of neutrophil apoptosis. These data shed important insights into the hitherto unrecognized importance of mitochondria in the function of neutrophils during infection and inflammation.

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Section: Discussionmentioning

confidence: 99%

The Mitochondrial Network of Human Neutrophils: Role in Chemotaxis, Phagocytosis, Respiratory Burst Activation, and Commitment to Apoptosis

Fossati

Moulding

Spiller

et al. 2003

The Journal of Immunology

308

251

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“…Consistent with this, we showed that the inhibition of NADPH oxidase via three distinctly different inhibitors resulted in an attenuation of extracellular peroxide levels in MZ-treated cells. DPI, a non-specific NADPH oxidase inhibitor, prevents electron transport within the NADPH oxidase multisubunit complex (O'Donnell et al, 1993;Li and Trush, 1998;Doussiere et al, 1999). Because of its action, however, DPI can also inhibit other electron transporters, like nitric oxide synthase (Stuehr et al, 1991), xanthine oxidase (Doussiere and Vignais, 1992), and mitochondrial complex I (Li and Trush, 1998).…”

Section: Discussionmentioning

confidence: 99%

“…DPI, a non-specific NADPH oxidase inhibitor, prevents electron transport within the NADPH oxidase multisubunit complex (O'Donnell et al, 1993;Li and Trush, 1998;Doussiere et al, 1999). Because of its action, however, DPI can also inhibit other electron transporters, like nitric oxide synthase (Stuehr et al, 1991), xanthine oxidase (Doussiere and Vignais, 1992), and mitochondrial complex I (Li and Trush, 1998). APO, which needs activation by peroxidases, inhibits the association of the NADPH oxidase cytosolic subunits (Stolk et al, 1994).…”

Section: Discussionmentioning

confidence: 99%

“…When NADPH oxidase subunits assemble on the cell membrane they can catalyze the electron transfer from NADPH to intermediate ligands, like PQ, which then redox cycle and generate ROS Zhang et al, 2004;BonnehBarkay, 2005b). To investigate the potential involvement of cellular oxidases, like NADPH oxidase, in the extracellular ROS generation by MZ, cultures were pretreated with several different NADPH oxidase inhibitors with differing mechanisms of action (O'Donnell et al, 1993;Stolk et al, 1994;Megyeri et al, 1995;Diatchuk et al, 1997;Li et al, 1998;Doussiere et al, 1999). DPI, apocynin and AEBSF all significantly attenuated H 2 O 2 generation in cultures treated with 30 uM MZ (Figs.…”

Section: Ros Generation By Mz May Involve Redox Cycling With Cellularmentioning

confidence: 99%

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Reactive oxygen species generation by the ethylene-bis-dithiocarbamate (EBDC) fungicide mancozeb and its contribution to neuronal toxicity in mesencephalic cells

Domico

Cooper

Bernard³

et al. 2007

NeuroToxicology

121

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Previous in vitro studies in our laboratory have shown that mancozeb (MZ) and maneb (MB), both widely used EBDC fungicides, are equipotent neurotoxicants that produce cell loss in mesencephalic dopaminergic and GABAergic cells after an acute 24 h exposure. Mitochondrial uncoupling and inhibition were associated with fungicide exposure. Inhibition of mitochondrial respiration is known to increase free radical production. Here the mechanism(s) of neuronal damage associated with MZ exposure was further explored by determining the role that reactive oxygen species (ROS) played in toxicity. Damage to mesencephalic dopamine and GABA cell populations were significantly attenuated when carried out in the presence of ascorbate or SOD indicative of a free radical mediated contribution to toxicity. ROS generation monitored by H 2 O 2 production using Amplex Red increased in a dose-dependent manner in response to MZ. Inhibition of intracellular catalase with aminotriazole had little effect on H 2 O 2 generation, whereas exogenously added catalase significantly reduced H 2 O 2 production demonstrating a large extracellular contribution to ROS generation. Conversely, cells preloaded with the ROS indicator dye DCF showed significant MZ-induced ROS production, demonstrating an increase in intracellular ROS. Both the organic backbone of MZ as well as its associated Mn ion, but not Zn ion were responsible and required for H 2 O 2 generation. The functionally diverse NADPH oxidase inhibitors, diphenylene iodonium chloride, apocynin, and 4-(2-aminoethyl)benzene-sulfonyl fluoride hydrochloride significantly attenuated H 2 O 2 production by MZ. In growth medium lacking cells, MZ produced little H 2 O 2 , but enhanced H 2 O 2 generation when added with xanthine plus xanthine oxidase whereas, in cultured cells, allopurinol partially attenuated H 2 O 2 production by MZ. Minocycline, an inhibitor of microglial activation, modestly reduced H 2 O 2 formation in mesencephalic cells. In contrast, neuronal enriched cultures or cultures treated with MAC-1-SAP to kill microglia, did not show an attenuation of ROS production. These findings demonstrate that Mn-containing EBDC fungicides such as MZ and MB can produce robust ROS generation that likely occurs via redox cycling with extracellular and intracellular oxidases. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. NIH Public Access Author ManuscriptNeurotoxicology. Author manuscript; available in PMC 2008 November 1. The findings further show that microglia may contribute to but are not required for ROS production by MZ.

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“…All effects by LPS could be blocked by diphenylene iodonium (DPI) but the four potential sources of • O 2 -, xanthine oxidase, NADPH oxidase, mitochondria and uncoupled NO synthase, will be inhibited by this unspecific flavoprotein inhibitor (Li and Trush, 1998;Rand and Li, 1993). L-NMMA as a blocker for NO-synthase oxidase would also block NO synthesis and specific inhibitors for its oxidase function do not exist to our knowledge.…”

Section: Xanthine Oxidase Is a Possible O 2 -Source After Lps Challengementioning

confidence: 99%

Superoxide as a Messenger of Endothelial Function

Ullrich

Bachschmid²

2000

Biochemical and Biophysical Research Communications

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Diphenyleneiodonium, an NAD(P)H Oxidase Inhibitor, also Potently Inhibits Mitochondrial Reactive Oxygen Species Production

Cited by 426 publications

References 17 publications

The Mitochondrial Network of Human Neutrophils: Role in Chemotaxis, Phagocytosis, Respiratory Burst Activation, and Commitment to Apoptosis

The Mitochondrial Network of Human Neutrophils: Role in Chemotaxis, Phagocytosis, Respiratory Burst Activation, and Commitment to Apoptosis

Reactive oxygen species generation by the ethylene-bis-dithiocarbamate (EBDC) fungicide mancozeb and its contribution to neuronal toxicity in mesencephalic cells

Superoxide as a Messenger of Endothelial Function

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