Mechanism of Menadione-Induced Cytotoxicity in Rat Platelets

BACKGROUND: Anti-ischemic therapy with organic nitrates as nitric oxide (NO) donors is complicated by the induction of tolerance. When nitrates are metabolized to release NO, there is a considerable coproduction of reactive oxygen species (superoxide radical and peroxynitrite) in vessels leading to inactivation of NO, to diminished cyclic quanosine monophosphate production in smooth muscle cells (SMC), to impaired vasomotor responses to the endothelium-derived relaxation factor (EDRF), and to formation of nitrotyrosine as a marker of glyceryltrinitrate (GTN)-induced formation of peroxynitrite. The aim of the study was to analyze in vitro the formation of superoxide radicals and of peroxynitrite in GTN-treated endothelial and smooth muscle cells and in washed ex vivo platelets using electron spin resonance and spin-trapping techniques. METHODS AND RESULTS: Using 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) as a spin trap, it was shown that in platelets, smooth muscle, and endothelial cells incubated acutely for 15 minutes with 0.5 mM GTN, the rate of generation of reactive oxygen species (ROS) was twice as high as under control conditions. Using the new spin-trap 2H-imidazole-1-oxide (TMIO), a GTN-induced peroxynitrite formation was detected in SMC and in platelets incubated with 0.5 mM GTN for 15 minutes. Spin-trap 1-hydroxy-3-carboxy-pyrrolidine (CP-H) was used to estimate the rate of ROS formation in platelets incubated for 15 minutes with 0.5 mM GTN; the rate amounted to 14.6 +/- 1.1 nM/min/mg protein compared with 4.0 +/- 0.4 nM/min/mg protein in controls. The rate of ROS formation in SMCs was substantially increased (240 +/- 16%) after initiation of GTN tolerance by treatment of the cells in culture with 100 µM GTN for 24 hours. CONCLUSIONS: GTN increases the formation of superoxide radicals in endothelial cells, SMCs, and platelets. Peroxynitrite is formed during GTN metabolism in vascular cells and may contribute to the development of tolerance. A decrease in the nitrate-induced inhibition of platelet aggregation during GTN tolerance is associated with oxidative actions of ROS formed in platelets during GTN metabolism.

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“…By increasing the activity of NADH oxidases, the formation of O2':' in platelets can be augmented (59).…”

Section: Discussionmentioning

confidence: 99%

Formation of Reactive Oxygen Species in Various Vascular Cells During Glyceryltrinitrate Metabolism

Dikalov¹,

Fink

Skatchkov

et al. 1998

J Cardiovasc Pharmacol Ther

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“…It can be produced in vivo by a number of oxidases (NADPH oxidases family [7] and xanthine oxidase [8]), by the mitochondrial electron transport chain [9], or by redox cycling agents (e.g., paraquat and menadione) in the presence of electron donors [10,11]. In a protic environment, O 2 •− undergoes spontaneous ( k = 2 × 10 5 M −1 s −1 ) [12] or SOD-catalyzed dismutation ( k = 1.6 × 10 9 M −1 s −1 ) [13] to form O 2 and H 2 O 2 .…”

Section: Introductionmentioning

confidence: 99%

Toward selective detection of reactive oxygen and nitrogen species with the use of fluorogenic probes – Limitations, progress, and perspectives

Dębowska

Dębski

Hardy

et al. 2015

Pharmacological Reports

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Over the last forty years, there has been tremendous progress in understanding the biological reactions of reactive oxygen species (ROS) and reactive nitrogen species (RNS). It is widely accepted that the generation of ROS and RNS is involved in physiological and pathophysiological processes. To understand the role of ROS and RNS in a variety of pathologies, the specific detection of ROS and RNS is fundamental. Unfortunately, the intracellular detection and quantitation of ROS and RNS remains a challenge. In this short review, we have focused on the mechanistic and quantitative aspects of their detection with the use of selected fluorogenic probes. The challenges, limitations and perspectives of these methods are discussed.

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“…Furthermore, thrombin-induced Ca 2þ increase was not affected by MPP þ . These results were confirmed by using a positive control, menadione, which is known to decrease the agonist-induced Ca 2þ increase [21].…”

Section: Effect Of Mpp þ On Platelet Lysis Total Glutathione (Gsh) Lmentioning

confidence: 58%

Inhibition of platelet aggregation by 1-methyl-4-phenyl pyridinium ion (MPP⁺) through ATP depletion: Evidence for the reduced platelet activities in Parkinson's disease

et al. 2009

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Neuronal accumulation of 1-methyl-4-phenylpyridinium ion (MPP(+)), the metabolite of neural toxin, 1-methyl-4-phenyl-1,2,3,6-tetrahyropyridine (MPTP), induces a rapid depletion of cellular ATP level and loss of neuronal cell viability which simulates human Parkinson's disease (PD). Since ATP plays an important role in the physiology and function of platelets, which share many biochemical and physiological features with neuronal cells, we examined the effect of MPP(+) on platelet aggregation and viability using freshly isolated rat platelets. While the treatment of MPP(+) to platelets did not induce cytotoxicity, it significantly attenuated agonist-induced platelet aggregation in a concentration dependent manner. The inhibition of aggregation by MPP(+) was mediated by the depletion of the cytoplasmic ATP pool and resultant decreased ATP secretion. Different from the previous reports in neuronal cells, MPP(+) did not affect intracellular levels of glutathione and cytoplasmic Ca(2+) in platelets. The combined treatment with MPP(+) and 2-deoxyglucose, a glycolysis inhibitor, showed the additive effect in the decrease of ATP secretion and intracellular content. Consistent with these findings, inhibitory effects of MPP(+) on platelet aggregation was significantly enhanced by the treatment with 2-deoxyglucose. In conclusion, these results suggested that MPP(+) can induce ATP depletion in platelets and attenuate platelet aggregation providing a new theory on the reduced platelet activities in PD patients.

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Mechanism of Menadione-Induced Cytotoxicity in Rat Platelets

Cited by 27 publications

References 0 publications

Formation of Reactive Oxygen Species in Various Vascular Cells During Glyceryltrinitrate Metabolism

Formation of Reactive Oxygen Species in Various Vascular Cells During Glyceryltrinitrate Metabolism

Toward selective detection of reactive oxygen and nitrogen species with the use of fluorogenic probes – Limitations, progress, and perspectives

Inhibition of platelet aggregation by 1-methyl-4-phenyl pyridinium ion (MPP⁺) through ATP depletion: Evidence for the reduced platelet activities in Parkinson's disease

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Mechanism of Menadione-Induced Cytotoxicity in Rat Platelets

Cited by 27 publications

References 0 publications

Formation of Reactive Oxygen Species in Various Vascular Cells During Glyceryltrinitrate Metabolism

Formation of Reactive Oxygen Species in Various Vascular Cells During Glyceryltrinitrate Metabolism

Toward selective detection of reactive oxygen and nitrogen species with the use of fluorogenic probes – Limitations, progress, and perspectives

Inhibition of platelet aggregation by 1-methyl-4-phenyl pyridinium ion (MPP+) through ATP depletion: Evidence for the reduced platelet activities in Parkinson's disease

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Product

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Inhibition of platelet aggregation by 1-methyl-4-phenyl pyridinium ion (MPP⁺) through ATP depletion: Evidence for the reduced platelet activities in Parkinson's disease