Cytoprotective effect of rhamnetin on miconazole-induced H9c2 cell damage.
- Author:
Kang Pa LEE
1
;
Jai Eun KIM
;
Won Hwan PARK
Author Information
- Publication Type:Original Article
- Keywords: Rhamnetin; miconazole; cardiomyocyte; apoptosis; APE/Ref-1
- MeSH: Antioxidants; Apoptosis; Caspase 3; Cell Death; Cell Proliferation; Cell Survival; Flow Cytometry; Heart; Hydrogen Peroxide; Miconazole; Myocytes, Cardiac; NADPH Oxidase; Reactive Oxygen Species; Real-Time Polymerase Chain Reaction; Up-Regulation
- From:Nutrition Research and Practice 2015;9(6):586-591
- CountryRepublic of Korea
- Language:English
- Abstract: 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 WelCount(TM) 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 microM) induced abnormal morphological changes and cell death in H9c2 cells. Rhamnetin enhanced the viability of miconazole (3 microM)-treated cells in a dose-dependent manner. Rhamnetin (1 and 3 microM) 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.
