Kaczara 2020 6th EU-Cardioprotection Meeting Riga

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Kaczara P, Przyborowski K, Sitek B, Kurpinska A, Kus K, Stojak M, Chlopicki S (2020) Carbon monoxide affords anti-platelet effects through the inhibition of oxidative phosphorylation at complex IV and inhibition of glycolysis by NAD+ depletion. 6th EU-Cardioprotection Meeting 2020 Riga LV.

Link: EU-Cardioprotection

Kaczara P, Przyborowski K, Sitek B, Kurpinska A, Kus K, Stojak M, Chlopicki S (2020)

Event: 6th EU-Cardioprotection Meeting 2020 Riga LV

Carbon monoxide (CO) inhibits platelet aggregation, but the mechanism involved has not been defined. We investigated a possible involvement of bioenergetics in anti-platelet effects of CO.

The effects of CO-releasing molecule CORM-A1 on human platelets aggregation, bioenergetics and metabolism were analysed using, respectively, a dual channel aggregometer, Seahorse XFe technique and liquid chromatography tandem-mass spectrometry (LC-MS/MS)-based metabolomics.

CORM-A1-induced inhibition of platelet aggregation was accompanied by inhibition of mitochondrial respiration and glycolysis. The effects of CORM-A1 on mitochondrial respiration were proved by a decrease in the concentration of tricarboxylic acid cycle intermediates and oxidized nicotinamide adenine dinucleotide (NAD+), consistent with the inhibition of oxidative phosphorylation. The CORM-A1-induced inhibition of glycolysis was associated with an increase in concentrations of proximal (before glyceraldehyde-3-phosphate dehydrogenase (GAPDH) ), but not distal glycolysis metabolites, implying that CO delayed glycolysis at the level of GAPDH, however GAPDH activity was unaffected by CO. Furthermore, in the presence of exogenous pyruvate the CORM-A1-induced effects on platelet aggregation and glycolysis were abolished, and restored after the inhibition of lactate dehydrogenase, which enables cytosolic NAD+ regeneration, suggesting that glycolysis inhibition resulted from NAD+ depletion.

The anti-platelet effect of CO is mediated by the simultaneous inhibition of oxidative phosphorylation and glycolysis by the inhibition of cytochrome c oxidase and depletion of NAD+, respectively.

Bioblast editor: Plangger M


Affiliations and support

Jagiellonian Centre Experimental Therapeutics (JCET), Jagiellonian Univ, Krakow, Poland

This work was supported by The National Centre for Research and Development [STRATEGMED1/233226/11/NCBR/2015]