Distinct effects of intracellular vs. extracellular acidic pH on the cardiac metabolome during ischemia and reperfusion. (January 2023)

• Record Type: Journal Article
• Authors: Milliken, Alexander S.; Ciesla, Jessica H.; Nadtochiy, Sergiy M.; Brookes, Paul S.
• Is Part Of: Journal of molecular and cellular cardiology. Volume 174(2023)
• Page Start: 101
• Page End: 114
• Publication Date: 2023-01
• Subjects: Metabolism -- pH -- Acidosis -- Purines -- Hypoxia -- Ischemia; 5NT ecto-5′-nucleotidase -- A-pHEX Acidic extracellular pH -- ATP adenosine triphosphate -- BCECF-AM 2′-7′-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein, acetoxymethyl ester -- CHIA Control, Hypoxia, Ischemia, Acidic pHIN -- IR ischemia-reperfusion -- MCT monocarboxylate transporter -- meth-ADP adenosine 5′-(α, β-methylene) diphosphate -- NBC Na+/HCO3− cotransporter -- NHE Na+/H+ exchanger -- NMN Nicotinamide mononucleotide -- Ox-Phos oxidative phosphorylation -- pHEX extracellular pH -- pHIN intracellular (cytosolic) pH -- PT pore permeability transition pore -- ROS reactive oxygen species -- RPP rate x pressure product -- TCA tricarboxylic acid; Cardiology -- Periodicals; Heart Diseases -- Periodicals; Molecular Biology -- Periodicals; Cardiologie -- Périodiques; Cardiology; Electronic journals; Periodicals; 616.12
• DOI: 10.1016/j.yjmcc.2022.11.008
• Languages: English
• ISSNs: 0022-2828

Abstract

Abstract: Tissue ischemia results in intracellular pH (pHIN ) acidification, and while metabolism is a known driver of acidic pHIN, less is known about how acidic pHIN regulates metabolism. Furthermore, acidic extracellular (pHEX ) during early reperfusion confers cardioprotection, but how this impacts metabolism is unclear. Herein we employed LCMS based targeted metabolomics to analyze perfused mouse hearts exposed to: (i) control perfusion, (ii) hypoxia, (iii) ischemia, (iv) enforced acidic pHIN, (v) control reperfusion, and (vi) acidic pHEX (6.8) reperfusion. Surprisingly little overlap was seen between metabolic changes induced by hypoxia, ischemia, and acidic pHIN . Acidic pHIN elevated metabolites in the top half of glycolysis, and enhanced glutathione redox state. Meanwhile, acidic pHEX reperfusion induced substantial metabolic changes in addition to those seen in control reperfusion. This included elevated metabolites in the top half of glycolysis, prevention of purine nucleotide loss, and an enhancement in glutathione redox state. These data led to hypotheses regarding potential roles for methylglyoxal inhibiting the mitochondrial permeability transition pore, and for acidic inhibition of ecto-5′-nucleotidase, as potential mediators of cardioprotection by acidic pHEX reperfusion. However, neither hypothesis was supported by subsequent experiments. In contrast, analysis of cardiac effluents revealed complex effects of pHEX on metabolite transport, suggesting that mildly acidic pHEX may enhance succinate release during reperfusion. Overall, each intervention had distinct and overlapping metabolic effects, suggesting acidic pH is an independent metabolic regulator regardless which side of the cell membrane it is imposed. Graphical abstract: Unlabelled Image Highlights: Hypoxia, ischemia and acidic pHIN each induce unique cardiac metabolic profiles. Acidic pHEX at reperfusion prevents purine loss and enhances succinate release. Surprisingly, acidic pHEX reperfusion has a greater metabolic impact than forced acid pHIN .