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Ganoderma atrum polysaccharide protects cardiomyocytes against anoxia/reoxygenation‐induced oxidative stress by mitochondrial pathway
Author(s) -
Li WenJuan,
Nie ShaoPing,
Chen Yi,
Xie MingYong,
He Ming,
Yu Qiang,
Yan Yan
Publication year - 2010
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.22526
Subject(s) - oxidative stress , cytochrome c , reactive oxygen species , apoptosis , superoxide dismutase , microbiology and biotechnology , mitochondrion , chemistry , intracellular , viability assay , cytosol , western blot , caspase 3 , biochemistry , biology , programmed cell death , enzyme , gene
It is now well established that oxidative stress plays a causative role in the pathogenesis of anoxia/reoxygenation (A/R) injury. Ganoderma atrum polysaccharide (PSG‐1), the most abundant component isolated from G. atrum , has been shown to possess potent antioxidant activity. The goals of this study were to investigate the effect of PSG‐1 against oxidative stress induced by A/R injury and the possible mechanisms in cardiomyocytes. In this work, primary cultures of neonatal rat cardiomyocytes pretreated with PSG‐1 were subjected to A/R and subsequently monitored for cell viability by the 3‐(4,5‐dimethyl‐2‐thiazolyl)‐2,5‐diphenyl‐2H‐tetrazolium bromide (MTT) assay. The levels of intracellular reactive oxygen species (ROS), apoptosis, and mitochondrial membrane potential (Δψ m ) were determined by flow cytometry. Western blot analysis was used to measure the expression of cytochrome c , Bcl‐2 family, and manganese superoxide dismutase (MnSOD) proteins, and the activities of caspase‐3 and caspase‐9 were determined by a colorimetric method. The results showed that PSG‐1 protected against cell death caused by A/R injury in cardiomyocytes. PSG‐1 reduced the A/R‐induced ROS generation, the loss of mitochondrial membrane potential (Δψ m ), and the release of cytochrome c from the mitochondria into cytosol. PSG‐1 inhibited the A/R‐stimulated activation of caspase‐9 and caspase‐3 and alteration of Bcl‐2 family proteins. Moreover, PSG‐1 significantly increased the protein expression of MnSOD in cardiomyocytes. These findings suggest that PSG‐1 significantly attenuates A/R‐induced oxidative stress and improves cell survival in cardiomyocytes through mitochondrial pathway. J. Cell. Biochem. 110: 191–200, 2010. © 2010 Wiley‐Liss, Inc.