Sevoflurane Attenuates Ischemia-Reperfusion Injury in a Rat Lung Transplantation Model Akihiro Ohsumi, MD, Katherine Marseu, MD, Peter Slinger, MD, Karen.

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Sevoflurane Attenuates Ischemia-Reperfusion Injury in a Rat Lung Transplantation Model Akihiro Ohsumi, MD, Katherine Marseu, MD, Peter Slinger, MD, Karen McRae, MD, Hyunhee Kim, MS, PhD, Zehong Guan, David M. Hwang, MD, Mingyao Liu, MS, MD, Shaf Keshavjee, MS, MD, Marcelo Cypel, MS, MD The Annals of Thoracic Surgery Volume 103, Issue 5, Pages 1578-1586 (May 2017) DOI: 10.1016/j.athoracsur.2016.10.062 Copyright © 2017 The Society of Thoracic Surgeons Terms and Conditions

Fig 1 Cytokine levels 2 hours after reperfusion. (A) Interleukin (IL)-1β in plasma, (B) IL-1β in lung tissue, (C) IL-6 in plasma, (D) IL-6 in lung tissue; (E) tumor necrosis factor (TNF)-α in plasma, (F) TNF-α in lung tissue. Data are shown as mean ± SD. The p values were calculated by one-way analysis of variance with Tukey’s multiple comparison tests. *p < 0.05. (PostC = postconditioning; PreC = preconditioning; PreC + PostC = both preconditioning and postconditioning.) The Annals of Thoracic Surgery 2017 103, 1578-1586DOI: (10.1016/j.athoracsur.2016.10.062) Copyright © 2017 The Society of Thoracic Surgeons Terms and Conditions

Fig 2 Myeloperoxidase (MPO) levels in (A) plasma and (B) lung tissue. Data are shown as mean ± SD. The p values were calculated by one-way analysis of variance with Tukey’s multiple comparison tests. *p < 0.05. (PostC = postconditioning; PreC = preconditioning; PreC + PostC = both preconditioning and postconditioning.) The Annals of Thoracic Surgery 2017 103, 1578-1586DOI: (10.1016/j.athoracsur.2016.10.062) Copyright © 2017 The Society of Thoracic Surgeons Terms and Conditions

Fig 3 Western blot analysis of (A) Bcl-2–associated X protein (Bax) and cytochrome c in mitochondria (loading control: voltage-dependent anion channel [VDAC]); and (B) Bax, cytochrome c, caspase-3, and cleaved caspase-3 in cytosol (loading control: β-actin); (C) caspase-3 cleavage ratio: cleaved caspase-3/caspase-3; (D) cytochrome c release: cytochrome c in cytosol/β-actin in cytosol. Data are shown as mean ± SD. The p values were calculated by Kruskal-Wallis test with Dunn’s multiple comparison tests. *p < 0.05. (PostC = postconditioning; PreC = preconditioning; PreC + PostC = both preconditioning and postconditioning.) The Annals of Thoracic Surgery 2017 103, 1578-1586DOI: (10.1016/j.athoracsur.2016.10.062) Copyright © 2017 The Society of Thoracic Surgeons Terms and Conditions

Fig 4 TUNEL staining for the (A) control, (B) PreC, (C) PostC, (D) PreC + PostC groups. Red dots are the apoptotic cells. (Magnification: ×200; scale bar = 200 μm.) (PostC = postconditioning; PreC = preconditioning; PreC + PostC = both preconditioning and postconditioning; TUNEL = terminal deoxynucleotidyl transferase-mediated deoxyuridinetriphosphate nick-end labeling.) The Annals of Thoracic Surgery 2017 103, 1578-1586DOI: (10.1016/j.athoracsur.2016.10.062) Copyright © 2017 The Society of Thoracic Surgeons Terms and Conditions

Fig 5 Hematoxylin and eosin staining for the (A) control, (B) PreC, (C) PostC, and (D) PreC + PostC groups and (E) Lung injury score. Arrowheads represent mononuclear cell infiltration. Bar indicates median value. The p values were calculated by Kruskal-Wallis test with Dunn’s multiple comparison tests. *p < 0.05. (PostC = postconditioning; PreC = preconditioning; PreC + PostC = both preconditioning and postconditioning.) The Annals of Thoracic Surgery 2017 103, 1578-1586DOI: (10.1016/j.athoracsur.2016.10.062) Copyright © 2017 The Society of Thoracic Surgeons Terms and Conditions

Supplemental Fig 1 The Annals of Thoracic Surgery 2017 103, 1578-1586DOI: (10.1016/j.athoracsur.2016.10.062) Copyright © 2017 The Society of Thoracic Surgeons Terms and Conditions

Supplemental Fig 2 The Annals of Thoracic Surgery 2017 103, 1578-1586DOI: (10.1016/j.athoracsur.2016.10.062) Copyright © 2017 The Society of Thoracic Surgeons Terms and Conditions