The Effect of Hypothermia on Lung Inducible Nitric Oxide Synthase Gene Expression in Intestinal Ischemia-Reperfusion Injury

장 허혈-재관류에서 폐조직의 Inducible Nitric Oxide Synthase유전자 발현에 대한 저체온증의 효과

  • Kim, Kyuseok (Department of Emergency Medicine, Seoul National University College of Medicine) ;
  • Lee, Jeong Hun (Department of Emergency Medicine, Seoul National University College of Medicine) ;
  • Suh, Gil Joon (Department of Emergency Medicine, Seoul National University College of Medicine) ;
  • Youn, Yeo Kyu (Department of Surgery, Seoul National University College of Medicine) ;
  • Kang, Young Joon (Department of Emergency Medicine, Cheju National University College of Medicine) ;
  • Kim, Min A (Department of Pathology, Seoul National University College of Medicine) ;
  • Cho, Sang-Gi (Department of Oral and Maxillofacial Surgery, School of Dentistry, Chonbuk National University) ;
  • Shin, Hyo-Keun (Department of Oral and Maxillofacial Surgery, School of Dentistry, Chonbuk National University)
  • 김규석 (서울대학교 의과대학 응급의학과) ;
  • 이정훈 (서울대학교 의과대학 응급의학과) ;
  • 서길준 (서울대학교 의과대학 응급의학과) ;
  • 윤여규 (서울대학교 의과대학 외과) ;
  • 강영준 (제주대학교 의과대학 응급의학과) ;
  • 김민아 (서울대학교 의과대학 병리학교실) ;
  • 조상기 (전북대학교 치과대학 구강악안면외과학교실) ;
  • 신효범 (전북대학교 치과대학 구강악안면외과학교실)
  • Received : 2006.05.12
  • Accepted : 2006.06.14
  • Published : 2006.06.30

Abstract

Purpose: Although hypothermia has been used in many clinical situations, such as post cardiopulmonary resuscitation, stroke, traumatic brain injury, septic shock, and hemorrhagic shock, the mechanism by which it works has not been clearly elucidated. We aimed to evaluate the effect of hypothermia on the plasma nitric oxide (NO) concentration, lung iNOS expression, and histologic changes in intestinal ischemia-reperfusion (IR). Method: Male Sprague-Dawley rats were randomly divided into the hypothermia group (HT, n=8, $27{\sim}30^{\circ}C$) and the normothermia group (NT, n=8, $36{\sim}37^{\circ}C$). They underwent 30 min of intestinal ischemia by clamping the superior mesenteric artery, which was followed by 1.5 h of reperfusion. They were then sacrificed. The acute lung injury (ALI) score, the plasma NO concentration, and lung iNOS gene expression were measured. Results: Compared with the HT group, the NT group showed severe infiltrations of inflammatrory cells, alveolar hemorrhages, and interstitial hypertrophies in lung tissues. There were significant differences in the ALI scores between the NT and the HT groups ($8.7{\pm}1.5/HPF$ in NT vs $5.8{\pm}1.2/HPF$ in HT, p=0.008). Although the plasma NO concentration was slightly lower in the HT group, there was no significant difference between the two groups ($0.80{\pm}0.24{\mu}mol/L$ in NT vs $0.75{\pm}0.30{\mu}mol/L$ in HT, p=0.917). Lung iNOS gene expression was stronger in the NT group than in the HT group. The band density of the expression of iNOS in lung tissues was significantly increased in the NT group compared to the HT group ($5.54{\pm}2.75$ in NT vs$0.08{\pm}0.52$ in HT, p=0.002). Conclusions: This study showed that hypothermia in intestinal IR reduces inflammatory responses, ALI scores, and iNOS gene expression in lung tissues. There was no significant effect of hypothermia on the plasma NO concentration.

Keywords

References

  1. Granger DN, Rutili G, McCord JM. Superoxide radicals in feline intestinsa ischemia. Gastroenteroloy 1981;81:22-9
  2. Crissinger RL, Dahlgren DN. Mucosal injury induced by ischemia and reperfusion in the piglet intestine: influence of age and feeding. Gastroenterology 1989;97:920-26 https://doi.org/10.1016/0016-5085(89)91498-4
  3. Weiss SJ. Oxygen, ischemia and inflammation. Acta Physiol Scand, Suppl 1986;548:9-37
  4. Engler RL, Dahlgren DD, Morris MA, et al. Role of leukocytes in response to acuta myocardial ischemia and reflow in dogs. Am J Physiol 1986;251:H314-H322
  5. Kanwar S, Tepperman BL, Payne D et al. Time course of nitric oxide production and epithelial dysfunction during ischemia/reperfusion of the feline small intestine. Circ Shock 1994;42:135-40
  6. Ulrich F, Hartmut K. Nitric oxide synthase: expression and expressional control of the three isoforms. Naunyn-Schmiedeberg's Arch Pharm 1995;352:351-364
  7. Robbins RA, Grisham MB. Nitric Oxide. Int J Biochem Cell Biol 1997;29:857-90 https://doi.org/10.1016/S1357-2725(96)00167-7
  8. Shigematsu K, Hamada Y, Hioki K, et al. Participation of nitric oxide in the mucosal injury of rat intestine induced by ischemia-reperfusion. J Pharmocol Exp Ther 1998;287:403-7
  9. Kubes P. Ischemia-reperfusion in feline small intestine: a role for nitric oxide. Am J Physiol 1993;264:G143-9
  10. Payne D, Kubes P. Nitric oxide donors reduce the rise in reperfusion-induced intestinal mucosal permeability. Am J Physiol 1993;265:G189-95
  11. Bernard SA, Gray TW, Buist MD, et al: Treatment of comatose survivors of out-of-hospital cardiac arrest with induced hypothermia. N Engl J Med 2002; 346:557-63 https://doi.org/10.1056/NEJMoa003289
  12. Conroy BP, Grafe MR, Jenkins LW, Vela AH, Lin CY, DeWitt DS, and Johnston WE. Histopathologic consequences of hyperglycemic cerebral ischemia during hypothermic cardiopulmonary bypass in pigs. Ann Thorac Surg 2001;71:1325-34 https://doi.org/10.1016/S0003-4975(01)02401-8
  13. Biagas KV and Gaeta ML. Treatment of traumatic brain injury with hypothermia. Curr Opin Pediatr 1998;10:271-77 https://doi.org/10.1097/00008480-199806000-00009
  14. Milde LN Clinical use of mild hypothermia for brain protection: a dream revisited. J Neurosurg Anesthesiol 1992;4:211-5 https://doi.org/10.1097/00008506-199207000-00012
  15. Ning XH, Chen SH, Xu CS, Li L, Yao LY, Qian K, Krueger JJ, Hyyti OM, Portman MA Hypothermic protection of the ischemic heart via alterations in apoptotic pathways as assessed by gene array analysis. J Appl Physiol 2002;92:2200-7 https://doi.org/10.1152/japplphysiol.01035.2001
  16. Kimura A, Sakurada S, Ohkuni H, Todome Y, Kurata K (2002) Moderate hypothermia delays proinflammatory cytokine production of human peripheral blood mononuclear cells. Crit Care Med 30:1499-502 https://doi.org/10.1097/00003246-200207000-00017
  17. Globus MY-T, Busto R, Lin B, Schnippering H, Ginsberg MD Detection of free radical activity during transient global ischemia and recirculation: effects of intraischemic brain temperature modulation. J Neurochem 1995;65:1250-6 https://doi.org/10.1046/j.1471-4159.1995.65031250.x
  18. Scumpia PO, Sarcia PJ, DeMarco VG, et al. Hypothermia attenuates iNOS, CAT-1, CAT-2, and nitric oxide expression in lungs of endotoxemic rats. Am J Physiol Lung Cell Mol Physiol 2002;283:L1231-L1238 https://doi.org/10.1152/ajplung.00102.2002
  19. Scumpia PO, Sarcia PJ, Kelly KM, et al. Hypothermia Induces Anti-Inflammatory Cytokines and Inhibits Nitric Oxide and Myeloperoxidase-Mediated Damage in the Hearts of Endotoxemic Rats. Chest 2004;125(4):1483-91 https://doi.org/10.1378/chest.125.4.1483
  20. Hassoun HT, Kozar RA, Kone BC, Safi HJ, Moore FA. Intraischemic hypothermia differentially modulates oxidative stress proteins during mesenteric ischemia/reperfusion. Surgery. 2002 Aug;132:369-76 https://doi.org/10.1067/msy.2002.125722
  21. Green LC, Wagner DA, Glogowski J, et al. Analysis of nitrate, nitrite, and nitrate in biological fluids. Anal Biochem 1982;126:131-138 https://doi.org/10.1016/0003-2697(82)90118-X
  22. Pierro A, Eaton S. Intestinal ischemia reperfusion injury and multisystem organ failure. Semin Pediatr Surg. 2004;13:11-7 https://doi.org/10.1053/j.sempedsurg.2003.09.003
  23. Parkinson EJ, Townsend PA, Stephanou A, Latchaman DS, Eaton S, Pierro A. The protective effect of moderate hypothermia during intestinal ischemia-reperfusion is associated with modification of hepatic transcription factor activation. J Pediatr Surg. 2004;39:696-701 https://doi.org/10.1016/j.jpedsurg.2004.01.025
  24. Kilbourn RG, Jubran J, Sazbo C, et al. Reversal of endotoxin-mediated shock by NG-methyl-L-arginine, and inhibitor of nitric oxide synthesis. Biochem Biophys Res Commun 1990;172:1132-8 https://doi.org/10.1016/0006-291X(90)91565-A