Protection effect of nicotinamide on cardiomyoblast hypoxia/re-oxygenation injury: study of cellular mitochondrial metabolism

Abstract

Hypoxia/re-oxygenation (H/R) injury is an important cause of heart failure and results in a critical metabolism dysfunction. In this paper, the cytoprotective effect of the nicotinamide adenine dinucleotide (NAD) precursor nicotinamide was evaluated using an in vitro model of cardiac H/R injury. Nicotinamide (0–20 mM) was applied to the myoblast cell line H9c2 which was subjected to hypoxia (12, 24, 36 h) followed by a re-oxygenation process (0, 4, 8, 12 h). Cell viability was measured, and mitochondrial metabolites were extracted and then measured by HPLC/MS/MS. The present study showed that nicotinamide could down-regulate the NADH/NAD ratio and then maintain the NAD-dependent metabolism processes. Furthermore, an aberrant decrease of fumarate levels and an increase of succinate levels were observed in the nicotinamide group, which was demonstrated to be caused by nicotinamide-induced succinate dehydrogenase (SDH) inhibition. These results suggest that nicotinamide exerts a protective effect on cardiomyoblasts against H/R-induced injury through both NADH/NAD regulation and reduction of reactive oxygen species generation via SDH inhibition.