Neobavaisoflavone Protects H9c2 Cells Against H2O2‐Induced Mitochondrial Dysfunction Through ALOX15/PGC1‐α Axis

ABSTRACT Neobavaisoflavone (NBIF) is a natural antioxidant that has a variety of pharmacological activities. To investigate the effects of NBIF on oxidative stress‐induced myocardial injury, H9c2 cells were treated with H 2 O 2 . Cell counting kit‐8 was used to detect cell viability. Intracellular as well as lipid radicals were detected. To measure mitochondrial function, tetramethylrhodamine ethyl ester was used to detect mitochondrial membrane potential. 12‐ and 15‐hydroxyeicosatetraenoic acids (HETE) were measured by LC‐MS/MS. ALOX15, which is the upstream protein of 12‐, 15‐HETE, was also measured by using western blot analysis. The results showed that H 2 O 2 induced lipid peroxidation in cardiomyocytes and caused mitochondrial dysfunction which was relieved by NBIF treatment. Besides, H 2 O 2 significantly increased the production of 12‐HETE and 15‐HETE and upregulated the expression of ALOX15 while PGC‐1α was downregulated and triggered the release of cytochrome c . The treatment of NBIF decreased the expression of ALOX15 and inhibited the activation of caspase‐3. NBIF protected mitochondrial membrane integrity through increasing PGC‐1α and Nrf1. Our results indicated that NBIF could protect cardiomyocytes against H 2 O 2 ‐induced mitochondrial dysfunction via ALOX15/PGC‐1α axis.