Therapeutic distant organ effects of regional hypothermia during mesenteric ischemia- reperfusion injury Rachel J. Santora, MD, Mihaela L. Lie, MD, Dmitry.

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Therapeutic distant organ effects of regional hypothermia during mesenteric ischemia- reperfusion injury Rachel J. Santora, MD, Mihaela L. Lie, MD, Dmitry N. Grigoryev, MD, PhD, Omer Nasir, MD, Frederick A. Moore, MD, Heitham T. Hassoun, MD Journal of Vascular Surgery Volume 52, Issue 4, Pages 1003-1014 (October 2010) DOI: 10.1016/j.jvs.2010.05.088 Copyright © 2010 Society for Vascular Surgery Terms and Conditions

Fig 1 Experimental model. A, Sprague-Dawley rats underwent sham laparotomy or superior mesenteric artery occlusion (SMAO) for 60 minutes with and without regional hypothermia. After the allotted reperfusion time, 6 or 24 hours, the animals were sacrificed and whole lung tissue was collected. The collected tissue was analyzed for evidence of lung injury and inflammation, and total RNA was isolated from the lung for gene expression profiling. B, Regional intraischemic hypothermia (RIH) is illustrated. For animals selected to undergo RIH, the intestines were exteriorized and placed between two cold compresses. A temperature probe was used to monitor RIH, and animals were placed on a heating pad to maintain systemic normothermia (37°C). Journal of Vascular Surgery 2010 52, 1003-1014DOI: (10.1016/j.jvs.2010.05.088) Copyright © 2010 Society for Vascular Surgery Terms and Conditions

Fig 2 Effect of regional hypothermia on lung injury during mesenteric ischemia-reperfusion injury (IRI). Representative sections of lung tissue at 24 hours after superior mesenteric artery occlusion, with and without hypothermia (H) are shown. IRI-treated rats demonstrate increased neutrophil influx (arrowhead) and alveolar hemorrhage (arrow) not present in sham, sham-H, or IRI-H (hematoxylin and eosin stain; original magnification ×40). Journal of Vascular Surgery 2010 52, 1003-1014DOI: (10.1016/j.jvs.2010.05.088) Copyright © 2010 Society for Vascular Surgery Terms and Conditions

Fig 3 Effect of regional hypothermia (H) on lung bronchoalveolar lavage (BAL) protein during mesenteric ischemia-reperfusion injury (IRI). BAL protein concentration (mg/mL) at 24 hours was greater during IRI compared with sham and IRI-H. *P ≤ .05, n ≥ 5 rats/group. The range bars show the standard error of the mean. Journal of Vascular Surgery 2010 52, 1003-1014DOI: (10.1016/j.jvs.2010.05.088) Copyright © 2010 Society for Vascular Surgery Terms and Conditions

Fig 4 Effect of regional hypothermia on lung edema during mesenteric ischemia-reperfusion injury (IRI). Lung edema (wet/dry ratio) was increased in IRI compared with sham and IRI-hypothermia (H). *P ≤ .05, n ≥ 5 rats/group. The range bars show the standard error of the mean. Journal of Vascular Surgery 2010 52, 1003-1014DOI: (10.1016/j.jvs.2010.05.088) Copyright © 2010 Society for Vascular Surgery Terms and Conditions

Fig 5 Hierarchic cluster analysis of lung gene expression during mesenteric ischemia-reperfusion injury (IRI) is shown at 6 and 24 hours. Each column represents an experimental condition of the corresponding lung sample. Hierarchic clustering using Pearson's correlation determined five major clusters (blue triangles), of which two clusters demonstrated clear differences in gene expression between IRI and IRI-hypothermia (H) at 6 hours. Genes from these clusters are highlighted with yellow rectangles and are listed on the right. Red indicates up-regulation, and green indicates down-regulation of gene expression with respect to corresponding controls. Journal of Vascular Surgery 2010 52, 1003-1014DOI: (10.1016/j.jvs.2010.05.088) Copyright © 2010 Society for Vascular Surgery Terms and Conditions

Fig 6 Expression of lipocalin-2 and chemokine ligand 1 (CXCL-1) genes in the lung by Ilumina microarray and quantitative real-time reverse-transcription polymerase chain reaction (RT-PCR). The relative fold-change (at 6 hours) identified by microarray analysis (filled bars) and real-time RT-PCR (open bars) was calculated for IRI and IRI-H samples with respect to corresponding shams and is represented as mean ± standard error of the mean. *Significant changes in gene expression between sham and treatment groups by either unpaired t-test (real-time RT-PCR) or significance analysis of microarrays (SAM; microarray). P ≤ .05 or q < 5%, respectively, n ≥ 3 samples/group. Journal of Vascular Surgery 2010 52, 1003-1014DOI: (10.1016/j.jvs.2010.05.088) Copyright © 2010 Society for Vascular Surgery Terms and Conditions