N-acetylcysteine alleviates H2O2-induced damage via regulating the redox status of intracellular antioxidants in H9c2 cells

N-acetylcysteine (NAC) is a thiol-containing antioxidant that modulates the intracellular redox state. NAC can scavenge reactive oxygen species (ROS) and maintain reduced glutathione (GSH) levels, in order to protect cardiomyocytes from oxidative stress. The present study aimed to determine whether NAC protects cardiomyocytes from oxidative damage by regulating the redox status of intracellular antioxidant proteins. The results revealed that NAC pretreatment increased cell viability and inhibited the activation of caspase-3, -8 and -9 during hydrogen peroxide (H2O2)-induced oxidative stress in H9c2 cells. Furthermore, decreased ROS levels, and increased total and reduced GSH levels were detected in response to NAC pretreatment. Non-reducing redox western blotting was performed to detect the redox status of intracellular antioxidant proteins, including thioredoxin 1 (Trx1), peroxiredoxin 1 (Prx1), GSH reductase (GSR), and phosphatase and tensin homolog (PTEN). The results revealed that the reduced form of Trx1 was markedly increased, and the oxidized forms of Prx1, GSR and PTEN were decreased following NAC pretreatment. Furthermore, NAC pretreatment decreased H2O2-induced phosphorylation of apoptosis signal-regulating kinase 1, which depends on the redox state of Trx1, and increased H2O2-induced phosphorylation of protein kinase B, which is essential to cell survival. To the best of our knowledge, the present study is the first to reveal that NAC pretreatment may alleviate oxidation of intracellular antioxidant proteins to inhibit oxidative stress-induced cardiomyocyte apoptosis.

[1]  F. Fang,et al.  Hydrogen protects against hyperoxia-induced apoptosis in type II alveolar epithelial cells via activation of PI3K/Akt/Foxo3a signaling pathway. , 2018, Biochemical and biophysical research communications.

[2]  N. Leslie,et al.  The PTEN protein: cellular localization and post-translational regulation. , 2016, Biochemical Society transactions.

[3]  T. Das,et al.  Inhibition of AKT promotes FOXO3a-dependent apoptosis in prostate cancer , 2016, Cell Death and Disease.

[4]  Yuqiang Wang,et al.  SIRT1 Protects Against Oxidative Stress‐Induced Endothelial Progenitor Cells Apoptosis by Inhibiting FOXO3a via FOXO3a Ubiquitination and Degradation , 2015, Journal of cellular physiology.

[5]  N. Maulik,et al.  Thioredoxins in cardiovascular disease. , 2015, Canadian journal of physiology and pharmacology.

[6]  N. Tonks,et al.  Reactivation of oxidized PTP1B and PTEN by thioredoxin 1 , 2014, The FEBS journal.

[7]  M. Hampton,et al.  Peroxiredoxins as biomarkers of oxidative stress. , 2014, Biochimica et biophysica acta.

[8]  A. Holmgren,et al.  The thioredoxin antioxidant system. , 2014, Free radical biology & medicine.

[9]  C. Lillig,et al.  Thioredoxins, glutaredoxins, and peroxiredoxins--molecular mechanisms and health significance: from cofactors to antioxidants to redox signaling. , 2013, Antioxidants & redox signaling.

[10]  Haobo Li,et al.  N-Acetylcysteine and allopurinol up-regulated the Jak/STAT3 and PI3K/Akt pathways via adiponectin and attenuated myocardial postischemic injury in diabetes. , 2013, Free radical biology & medicine.

[11]  Michael Berk,et al.  The chemistry and biological activities of N-acetylcysteine. , 2013, Biochimica et biophysica acta.

[12]  A. Wen,et al.  Safflor yellow A protects neonatal rat cardiomyocytes against anoxia/reoxygenation injury in vitro , 2013, Acta Pharmacologica Sinica.

[13]  I. Benjamin,et al.  Glutathione‐dependent reductive stress triggers mitochondrial oxidation and cytotoxicity , 2012, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[14]  C. Harris,et al.  Oxidative stress, thiols, and redox profiles. , 2012, Methods in molecular biology.

[15]  C. L. Buller,et al.  Impact of N-Acetylcysteine on Neonatal Cardiomyocyte Ischemia-Reperfusion Injury , 2011, Pediatric Research.

[16]  N. Leslie,et al.  Prdx1 inhibits tumorigenesis via regulating PTEN/AKT activity , 2009, The EMBO journal.

[17]  Santosh Kumar,et al.  N-acetylcysteine prevents glucose/glucose oxidase-induced oxidative stress, mitochondrial damage and apoptosis in H9c2 cells. , 2009, Life sciences.

[18]  C. Myers,et al.  Acrolein oxidizes the cytosolic and mitochondrial thioredoxins in human endothelial cells. , 2008, Toxicology.

[19]  A. Holmgren,et al.  Glutaredoxin systems. , 2008, Biochimica et biophysica acta.

[20]  D. Yellon,et al.  Myocardial reperfusion injury. , 2007, The New England journal of medicine.

[21]  K. Otsu,et al.  The role of apoptosis signal-regulating kinase 1 in cardiomyocyte apoptosis. , 2006, Antioxidants & redox signaling.

[22]  Dean P. Jones,et al.  Nuclear and mitochondrial compartmentation of oxidative stress and redox signaling. , 2006, Annual review of pharmacology and toxicology.

[23]  A. Holmgren,et al.  Thiol redox control via thioredoxin and glutaredoxin systems. , 2005, Biochemical Society transactions.

[24]  L. Becker.  Myocardial Reperfusion Injury , 2004, Journal of Thrombosis and Thrombolysis.

[25]  Hajime Nakamura,et al.  Glutaredoxin Exerts an Antiapoptotic Effect by Regulating the Redox State of Akt* , 2003, Journal of Biological Chemistry.

[26]  C. Downes,et al.  Redox regulation of PI 3‐kinase signalling via inactivation of PTEN , 2003, The EMBO journal.

[27]  Yong J. Lee,et al.  Differential role of glutaredoxin and thioredoxin in metabolic oxidative stress-induced activation of apoptosis signal-regulating kinase 1. , 2003, The Biochemical journal.

[28]  M. Suntharalingam,et al.  Role of Glutaredoxin in Metabolic Oxidative Stress , 2002, The Journal of Biological Chemistry.

[29]  W. Min,et al.  Thioredoxin Promotes ASK1 Ubiquitination and Degradation to Inhibit ASK1-Mediated Apoptosis in a Redox Activity-Independent Manner , 2002, Circulation research.

[30]  M. Suntharalingam,et al.  Role of Glutaredoxin in Metabolic Oxidative Stress GLUTAREDOXIN AS A SENSOR OF OXIDATIVE STRESS MEDIATED BY H 2 O 2 * , 2002 .

[31]  Sue Goo Rhee,et al.  Hydrogen Peroxide: A Key Messenger That Modulates Protein Phosphorylation Through Cysteine Oxidation , 2000, Science's STKE.

[32]  Kohei Miyazono,et al.  Mammalian thioredoxin is a direct inhibitor of apoptosis signal‐regulating kinase (ASK) 1 , 1998, The EMBO journal.

[33]  M. Noble,et al.  N-acetyl-L-cysteine is a pluripotent protector against cell death and enhancer of trophic factor-mediated cell survival in vitro. , 1994, Proceedings of the National Academy of Sciences of the United States of America.