Protective effects of glutathione on oxidative injury induced by hydrogen peroxide in intestinal epithelial cells Haitao Ren, MD, PhD, a,b,1 Qinghe Meng, MD, a,1 Natesh Yepuri, MBBS, a Xianjin Du, MD, PhD, a James Osei Sarpong, BA, a and Robert N. Cooney, MD a, * a Department of Surgery, SUNY Upstate Medical University, Syracuse, New York b Department of Burns and Wound Center, The Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, China article info Article history: Received 1 June 2017 Received in revised form 21 August 2017 Accepted 28 September 2017 Available online xxx Keywords: GSH IEC-6 Apoptosis NF-kB P38 MAPK IL-1b ROS Hydrogen peroxide (H 2 O 2 ) abstract Background: Reactive oxygen species are increased in multiple gastrointestinal diseases and contribute to their pathogenesis. glutathione (GSH) is an antioxidant that helps to prevent reactive oxygen speciesemediated mucosal damage. This study examines the mechanisms by which GSH attenuates hydrogen peroxide (H 2 O 2 )einduced injury in intestinal epithelial cells. Methods: IEC-6 cells were cultured and treated with H 2 O 2  GSH. Inflammation was measured by nuclear factor kappa-B (NF-kB) P65 expression, NF-kB nuclear translocation, ikBa phosphorylation, and interleukin 1 beta secretion. Terminal deoxynucleotidyl trans- feraseemediated UTP end-labeling staining and cleaved caspase-3 were used to assess apoptosis. The role of P38 mitogen-activated protein kinase (P38 MAPK) signaling was examined using the P38 MAPK agonist U46619 and inhibitor SB203580 in H 2 O 2 and GSH- treated cells. Phosphorylated and total P38 MAPKs and cleaved caspase-3 were measured by Western blot. Data are means  standard deviation, statistical significance P < 0.05 by student’s t-test, or one-way analysis of variance. Results: Pretreatment with GSH attenuates the activation of NF-kB and P38 MAPK signaling pathways by H 2 O 2 . GSH also decreased H 2 O 2 -mediated increases in interleukin 1 beta secretion, cleaved caspase-3 activation, and apoptosis in IEC-6 cells. SB203580 attenuated the increase in apoptosis and cleaved caspase-3 in H 2 O 2 -treated cells. The increase in apoptotic index and cleaved caspase-3 observed in U46619-treated cells was also dimin- ished by GSH. Conclusions: GSH appears to ameliorate oxidative injury in intestinal epithelial cells by attenuating H 2 O 2 -mediated activation of NF-kB and P38 MAPK signaling pathways that regulate intestinal inflammation and apoptosis. ª 2017 Elsevier Inc. All rights reserved. * Corresponding author. Department of Surgery, Upstate Medical University, 750 E. Adams St, Suite 8141, Syracuse, NY 13210. Tel.: þ1 315 464 5549; fax: þ1 315 464 3649. E-mail address: cooneyr@upstate.edu (R.N. Cooney). 1 These authors contributed equally to this work. Available online at www.sciencedirect.com ScienceDirect journal homepage: www.JournalofSurgicalResearch.com journal of surgical research  february 2018 (222) 39 e47 0022-4804/$ e see front matter ª 2017 Elsevier Inc. All rights reserved. https://doi.org/10.1016/j.jss.2017.09.041