Resveratrol attenuates oxidant-induced mitochondrial damage in embryonic rat cardiomyocytes via inactivating GSK-3β
10.3760/cma.j.issn.0253-3758.2012.10.012
- VernacularTitle:糖原合成酶激酶3β在白藜芦醇诱导的大鼠心肌线粒体保护中的作用及其机制
- Author:
Yong-Gui HE
1
;
Yu-Jie SUN
;
Yu-Xi XIE
;
Huan ZHENG
;
Yi-Dong ZHANG
;
Jing GUO
;
Jin-Kun XI
Author Information
1. 河北联合大学医学实验研究中心心脏研究所
- Keywords:
Myocardium;
Mitochondria;
Heart;
Glycogen synthase kinase 3;
Resveratrol
- From:
Chinese Journal of Cardiology
2012;40(10):858-863
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the underlying mechanism of the protective effects of resveratrol on oxidant-induced mitochondrial damage in embryonic rat cardiomyocytes.Methods H9c2 cells,a permanent cell line derived from embryonic rat cardiac tissue,and then randomly divided into control group [PBS,cells exposed to H2O2 (600 μmol/L) for 20 min to induce mitochondrial oxidant damage],resveratrol group (0.01,0.1,1,5,10 and 20 μmol/L for 20 min at 20 min before exposing to H2O2),resveratrol plus inhibitor group (1 μmol/L KT5823 for 10 min at 10 min before 5 pmol/L resveratrol treatment) and inhibitor group (1 μmol/L KT5823 for 10 min).Mitochondrial membrane potential (△Ψm) was measured by staining cells with tetramethylrhodamine ethyl ester (TMRE) and the mitochondrial permeability transition pore (mPTP) opening was evaluated by measuring the decrease of TMRE fluorescence intensity.Immunofluorescence assay was used to observe GSK-3β phosphorylation.The phosphorylation of GSK-3β and VASP were determined by Western blot.To detect intracellular NO,cells were loaded with DAF-FM DA (specific fluorescent dye of NO) and imaged with confocal microscopy.Results Compared to the control group,resveratrol (0.01-5 μmol/L) attenuated H2O2-inducedmitochondrial damage reflected by attenuating the H2O2-induced TMRE fluorescence intensity decrease in a dose-dependent manner and the efficacy of 10 and 20 μmol/L resveratrol was significantly lower than that of 5 μmol/L resveratrol.Resveratrol also significantly upregulated the protein expression of VASP and increased GSK-3β Ser9 phosphorylation,which could lead the inactivation of GSK-3β.These effects of resveratrol could be significantly abolished by protein kinase G inhibitor KT5823,while KT5823 alone did not affect CSK-3β and VASP phosphorylation.Confocal microscopy showed that DAF-FM (specific NO indicator) was similar between resveratrol and control group,suggesting that resveratrol did not produce NO.Conclusions Resveratrol could attenuate oxidant-induced mitochondrial damage in embryonic rat cardiomyocytes by inactivating GSK-3β via cGMP/PKG signaling pathway independent of NO-related mechanism.
