Cytoprotective effect of rhamnetin on miconazole-induced H9c2 cell damage

• Published in: Nutrition research and practice 2015, 9(6), , pp.586-591
• Main Authors: Lee, K.P., Konkuk University, Seoul, Republic of Korea, Kim, J.E., Dongguk University, Goyang, Republic of Korea, Park, W.H., Dongguk University, Goyang, Republic of Korea
• Format: Article
• Language: English
• Subjects: APOPTOSE; APOPTOSIS; Original Research; Rhamnetin,miconazole,cardiomyocyte,APE/Ref-1; 생활과학
• ISSN: 1976-1457; 2005-6168
• DOI: 10.4162/nrp.2015.9.6.586

BACKGROUND/OBJECTIVES: Reactive oxygen species (ROS) formation is closely related to miconazole-induced heart dysfunction. Although rhamnetin has antioxidant effects, it remained unknown whether it can protect against miconazole-induced cardiomyocyte apoptosis. Thus, we investigated the effects of rhamnetin on miconazole-stimulated H9c2 cell apoptosis. MATERIALS/METHODS: Cell morphology was observed by inverted microscope and cell viability was determined using a WelCountTM cell proliferation assay kit. Miconazole-induced ROS production was evaluated by fluorescence-activated cell sorting with 6-carboxy-2 ,7-dichlorofluoroscein diacetate (H2DCF-DA) stain. Immunoblot analysis was used to determine apurinic/apyrimidinic endonuclease 1 (APE/Ref-1) and cleaved cysteine-aspartic protease (caspase) 3 expression. NADPH oxidase levels were measured using real-time polymerase chain reaction. RESULTS: Miconazole (3 and 10 mu M) induced abnormal morphological changes and cell death in H9c2 cells. Rhamnetin enhanced the viability of miconazole (3 mu M)-treated cells in a dose-dependent manner. Rhamnetin (1 and 3 mu M) treatment downregulated cleaved caspase 3 and upregulated APE/Ref-1 expression in miconazole-stimulated cells. Additionally, rhamnetin significantly reduced ROS generation. CONCLUSIONS: Our data suggest that rhamnetin may have cytoprotective effects in miconazole-stimulated H9c2 cardiomyocytes via ROS inhibition. This effect most likely occurs through the upregulation of APE/Ref-1 and attenuation of hydrogen peroxide levels.