Pharmacological interventions that activate mitochondrial biogenesis stimulate nucleotide generation in...

Mitochondrial dysfunction in failing hearts has been described as a driving force for energy deprivation and cardiomyocyte energy supply-demand imbalance. Isoproterenol (ISO), the β1/β2-adrenergic receptor agonist that leads to myocardial stress and mitochondrial damage, is extensively used for in vitro and in vivo studies to test the efficacy of therapeutic strategies in heart failure (HF). This study evaluated the cell morphology, nucleotide concentrations, and mitochondrial function of ISO-treated cardiomyocytes stimulated with the activators of mitochondrial biogenesis and nucleotide precursors. H9c2 rat cardiomyocyte line cells were treated with ISO in the presence of mitochondrial biogenesis stimuli quercetin (Que), rosiglitazone (Ros), S-Nitroso-N-acetyl-DL-penicillamin (SNAP), and NAD⁺ precursor, nicotinamide riboside (NR). The intracellular concentrations of nucleotides were analyzed using high-performance liquid chromato-graphy, and the Seahorse metabolic flux analyzer determined the mitochondrial function. ISO decreased intracellular ATP concentration in H9c2 cells as compared to control. The treatment with SNAP increased ATP concentration compared to ISO-only treated cells, while Que, Ros, and NR had no effect. NR treatment led to the elevation of intracellular NAD⁺ concentration, while the treatment with SNAP, Ros, and NR stimulated the mitochondrial respiration in ISO-pretreated H9c2 cells. In conclusion, mitochondrial biogenesis activators consistently improved cardiomyocyte mitochondrial function, but only selected molecules helped to improve ATP or NAD⁺ concentrations. This information may help to optimize treatment to ameliorate energy imbalance in failing cardiomyocytes.