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24 March 2016 : Animal Research  

Puerarin Attenuates Anoxia/Reoxygenation Injury Through Enhancing Bcl-2 Associated Athanogene 3 Expression, a Modulator of Apoptosis and Autophagy

Yayu MaABCDEF, Ya GaiABCDE, Jingpeng YanBCD, Jian LiABC, Yangyang ZhangDEF

DOI: 10.12659/MSM.897379

Med Sci Monit 2016; 22:977-983

Abstract

BACKGROUND: Puerarin has protective effects on ischemia-reperfusion injury, but the underlying mechanisms are not fully revealed. This study explored the effect of puerarin on the expression of Bcl-2 associated athanogene 3 (BAG3) in an in vitro model of anoxia/reoxygenation injury (A/RI) in neonate rat primary cardiomyocytes and the functions of BAG3 in A/RI.

MATERIAL AND METHODS: BAG3 expression in cardiomyocytes with or without puerarin pre-treatment was quantified using qRT-PCR and Western blot analysis. The effects of BAG3 on A/RI were studied by measuring the activity of lactate dehydrogenase (LDH) and creatine phosphate kinase (CPK), the concentration of malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px). The effects of BAG3 on autophagy and apoptosis of the cardiomyocytes after A/RI were further studied.

RESULTS: Puerarin significantly promoted BAG3 expression in the rat primary cardiomyocytes after A/RI. Enforced BAG3 expression presented similar effects as puerarin pre-treatment in attenuating A/RI in terms of CPK, LDH, MDA, SOD, GSH-Px, ROS generation, and cell viability. BAG3 overexpression significantly stimulated autophagy in cardiomyocytes after A/RI, which presented protective effects on A/RI in terms of cell viability and apoptosis. Autophagy inhibition partly abrogated the protective effects of BAG3.

CONCLUSIONS: Puerarin can directly increase BAG3 transcription and translation in cardiomyocytes after A/RI. The elevated BAG3 expression presents protective effects on A/RI at least through enhancing autophagy and reducing apoptosis, which is a novel protective mechanism of puerarin in ARI.

Keywords: Adaptor Proteins, Signal Transducing - metabolism, Animals, Apoptosis - drug effects, Apoptosis Regulatory Proteins - metabolism, Autophagy - drug effects, Cell Hypoxia - drug effects, Cell Survival - drug effects, Cells, Cultured, Creatine Kinase - metabolism, Glutathione Peroxidase - metabolism, Isoflavones - pharmacology, L-Lactate Dehydrogenase - metabolism, Malondialdehyde - metabolism, Myocytes, Cardiac - pathology, Oxygen - pharmacology, Rats, Sprague-Dawley, Reactive Oxygen Species - metabolism, Superoxide Dismutase - metabolism

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750