Role of Toll-Like Receptor-4 in Lung Ischemia-Reperfusion Injury

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Role of Toll-Like Receptor-4 in Lung Ischemia-Reperfusion Injury Heather E. Merry, MD, Patrick Phelan, MD, Mathew R. Doak, BS, Minqing Zhao, MD, PhD, Billanna Hwang, MPH, Michael S. Mulligan, MD The Annals of Thoracic Surgery Volume 99, Issue 4, Pages 1193-1199 (April 2015) DOI: 10.1016/j.athoracsur.2014.12.062 Copyright © 2015 The Society of Thoracic Surgeons Terms and Conditions

Fig 1 Sequence-specific toll-like receptor (TLR)-4 knockdown with TLR-4 short interference (si)RNA. Pretreatment with TLR-4 siRNA prior to the ischemia and reperfusion (IR) protocol resulted in a 69% (p < 0.01) and 92% (p < 0.001) decrease in TLR4 concentration on western blot for sequences 1 and 2, respectively, when compared with animals receiving control non-coding siRNA. The Annals of Thoracic Surgery 2015 99, 1193-1199DOI: (10.1016/j.athoracsur.2014.12.062) Copyright © 2015 The Society of Thoracic Surgeons Terms and Conditions

Fig 2 Cell-specific toll-like receptor (TLR)-4 knockdown with TLR-4 short interference (si)RNA. Alveolar macrophages (AM) isolated from lungs treated with TLR-4 siRNA resulted in a 93% reduction (p < 0.001) in TLR-4 levels on western blot when compared with lungs treated with non-coding siRNA. No significant reduction in TLR-4 levels was found in pulmonary artery endothelial cells (PAEC), type 2 pneumocytes (T2P), or circulating white blood cells (WBC) isolated from animals treated with TLR-4 siRNA compared with those treated with non-coding siRNA. The Annals of Thoracic Surgery 2015 99, 1193-1199DOI: (10.1016/j.athoracsur.2014.12.062) Copyright © 2015 The Society of Thoracic Surgeons Terms and Conditions

Fig 3 Reduction on stress-activated protein kinase activation with toll-like receptor (TLR)-4 knockdown in lung ischemia-reperfusion injury. Although there is no difference between total levels of c-Jun-terminal kinase (JNK) and p38 after the ischemia and reperfusion (IR) protocol when comparing control and toll-like receptor (TLR)-4 short interference (si)RNA, the levels of phosphorylated JNK and p38 are significantly decreased (p < 0.01) for animals that were pretreated with TLR-4 siRNA compared with controls. The Annals of Thoracic Surgery 2015 99, 1193-1199DOI: (10.1016/j.athoracsur.2014.12.062) Copyright © 2015 The Society of Thoracic Surgeons Terms and Conditions

Fig 4 The toll-like receptor (TLR)-4 knockdown reduced permeability index in lung ischemia-reperfusion injury. The ischemia and reperfusion (IR) protocol increases vascular permeability ninefold in control animals pretreated with no short interference (si)RNA, TLR-4 siRNA prior to the IR protocol results in vascular permeability 90% less than positive controls (p < 0.001). This is equivalent to control animals that did not undergo the IR protocol. The Annals of Thoracic Surgery 2015 99, 1193-1199DOI: (10.1016/j.athoracsur.2014.12.062) Copyright © 2015 The Society of Thoracic Surgeons Terms and Conditions