A closer look into NADPH oxidase inhibitors: Validation and insight into their mechanism of action

Reis, Joana; Massari, Marta; Marchese, Sara; Ceccon, Marta; Aalbers, Friso S.; Corana, Federica; Valente, Sérgio; Mai, Antonello; Magnani, Francesca; Mattevi, A.

doi:10.1016/j.redox.2020.101466

Cited by 72 publications

(75 citation statements)

References 58 publications

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“…We used CATPB (500 nM), as an antagonist of the active site of the receptor, 27 , 28 genistein (100 nM) to inhibit the phosphorylation of Gα/q11 proteins, which is necessary for intracellular signaling of FFA2R, 28 and DPI (10 µM) as an inhibitor of the Heme groups and FAD of the NADPH oxidase complex. 29 We incubated the neutrophils with each inhibitor 1 h before the induction of NET with SCFAs as described in section “Stimulation of NET with SCFAs.”…”

Section: Methodsmentioning

confidence: 99%

Physiological concentrations of short-chain fatty acids induce the formation of neutrophil extracellular traps in vitro

Íñiguez-Gutiérrez

Godínez-Méndez

Fafutis‐Morris

et al. 2020

Int J Immunopathol Pharmacol

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Neutrophils represent the first line of host cellular defense against various pathogens. The most recently described microbicidal mechanism of these cells is the release of neutrophil extracellular traps (NET). Currently, a wide range of chemical and biological stimuli are known to induce this response; however, the effect of short-chain fatty acids (SCFAs) on the induction of NET is still unknown. SCFAs are produced mainly by bacterial fermentation of dietary fiber and are found in host tissues and blood. This study aimed to determine whether physiological levels of SCFAs can induce the formation of NET. Previously reported concentrations of SCFAs (as found in the colonic lumen and peripheral blood in postprandial and basal states) were used to stimulate the neutrophils. In order to determine the signaling pathway utilized by SCFAs, we tested the inhibition of the Free Fatty Acid 2 Receptor (FFA2R) expressed in neutrophils using CATPB, the inhibitor of FFA2R, genistein, an inhibitor of the downstream Gα/q11 proteins and DPI, an inhibitor of the NADPH oxidase complex. The SCFAs at colonic intestinal lumen concentrations were able to induce the formation of NET, and when tested at concentrations found in the peripheral blood, only acetic acid at 100 μM (fasting equivalent) and 700 μM (postprandial equivalent) was found to induce the formation of NET. The administration of the competitive inhibitor against the receptor or blockade of relevant G protein signaling and the inhibition of NADPH oxidase complex decreased NET release. SCFAs stimulate NET formation in vitro and this effect is mediated, in part, by the FFA2R.

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Section: Methodsmentioning

confidence: 99%

Physiological concentrations of short-chain fatty acids induce the formation of neutrophil extracellular traps in vitro

Íñiguez-Gutiérrez

Godínez-Méndez

Fafutis‐Morris

et al. 2020

Int J Immunopathol Pharmacol

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“…One of the main differences in these studies is that here we used mouse islets from wild type and mice knockout for NADPH oxidase 2, while in other studies, immortalized cell lines and protein knockdown or unspecific inhibitors of NADPH oxidase such as DPI were used. These inhibitors can act on different NADPH oxidases and also in other unspecific intracellular targets [52]. Nonetheless, we cannot exclude that other NADPH oxidases have impact on insulin secretion upon GPR40 activation.…”

Section: Discussionmentioning

confidence: 95%

Evidence for NADPH oxidase activation by GPR40 in pancreatic β-cells

et al. 2020

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Objective: Investigate the involvement of the fatty acids receptor GPR40 in the assembly and activation of NADPH oxidase and the implications on pancreatic β-cell function. Methods: BRIN-BD11 β-cells were exposed to GPR40 agonist (GW9508) or linoleic acid in different glucose concentrations. Superoxide and H 2 O 2 were analyzed, respectively, by DHE fluorescence and by fluorescence of the H 2 O 2 sensor, roGFP2-Orp1. Protein contents of p47 phox in plasma membrane and cytosol were analyzed by western blot. NADPH oxidase role was evaluated by p22 phox siRNA or by pharmacological inhibition with VAS2870. NOX2 KO islets were used to measure total cytosolic calcium and insulin secretion. Results: GW9508 and linoleic acid increased superoxide and H 2 O 2 contents at 5.6 and 8.3 mM of glucose. In addition, in 5.6 mM, but not at 16.7 mM of glucose, activation of GPR40 led to the translocation of p47 phox to the plasma membrane. Knockdown of p22 phox abolished the increase in superoxide after GW9508 and linoleic acid. No differences in insulin secretion were found between wild type and NOX2 KO islets treated with GW9508 or linoleic acid. Discussion: We report for the first time that acute activation of GPR40 leads to NADPH oxidase activation in pancreatic β-cells, without impact on insulin secretion.

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“…Thus, GKT136901 and GKT137831 proved to be strong NOX4 inhibitors [ 131 ]. However, some of these compounds are not specific for NOX isoforms as some of them are able to inhibit xanthine oxidase (XO), eNOS and an enzyme of mitochondrial complex 1 [ 132 ].…”

Section: Introductionmentioning

confidence: 99%

“…However, controversial results have been reported on the specificity of VAS2870 and about its possible inhibitory action in cell-free assays or in cell assays, for instance, it has been cited that VAS2870 inhibits NOX2 in whole-cell lysates and as a direct NOX2 inhibitor, while another study suggests that VAS2870 showed no inhibitory activity in NOX2 and NOX5 membrane assays [ 134 ]. VAS2870 displayed low to sustained (40–70%) inhibition against NOX2 and NOX4 but not against NOX5 [ 131 ], therefore, although VAS2870 could be a good candidate to be evaluated in AD models, is cytotoxic [ 135 ].…”

Section: Introductionmentioning

confidence: 99%

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Implication of Nicotinamide Adenine Dinucleotide Phosphate (NADPH) Oxidase and Its Inhibitors in Alzheimer’s Disease Murine Models

2021

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Alzheimer’s disease (AD) is one of the main human dementias around the world which is constantly increasing every year due to several factors (age, genetics, environment, etc.) and there are no prevention or treatment options to cure it. AD is characterized by memory loss associated with oxidative stress (OS) in brain cells (neurons, astrocytes, microglia, etc.). OS can be produced by amyloid beta (Aβ) protein aggregation and its interaction with metals, mitochondrial damage and alterations between antioxidants and oxidant enzymes such as nicotinamide adenine dinucleotide phosphate (NADPH) oxidase. NADPH oxidase produces reactive oxygen species (ROS) and it is overexpressed in AD, producing large amounts of superoxide anions and hydrogen peroxide which damage brain cells and the vasculature. In addition, it has been reported that NADPH oxidase causes an imbalance of pH which could also influence in the amyloid beta (Aβ) production. Therefore, NADPH oxidase had been proposed as a therapeutic target in AD. However, there are no drugs for AD treatment such as an NADPH oxidase inhibitor despite great efforts made to stabilize the ROS production using antioxidant molecules. So, in this work, we will focus our attention on NADPH oxidase (NOX2 and NOX4) in AD as well as in AD models and later discuss the use of NADPH oxidase inhibitor compounds in AD.

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A closer look into NADPH oxidase inhibitors: Validation and insight into their mechanism of action

Cited by 72 publications

References 58 publications

Physiological concentrations of short-chain fatty acids induce the formation of neutrophil extracellular traps in vitro

Physiological concentrations of short-chain fatty acids induce the formation of neutrophil extracellular traps in vitro

Evidence for NADPH oxidase activation by GPR40 in pancreatic β-cells

Implication of Nicotinamide Adenine Dinucleotide Phosphate (NADPH) Oxidase and Its Inhibitors in Alzheimer’s Disease Murine Models

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