Histopathological Changes at Colonic Anastomotic Site after Ischemia Reperfusion Injury: Role of Aminoguanidine in Experimental Model

Abstract

Background Inducible nitric oxide synthase (iNOS) has a significant role in ischemia reperfusion (I-R) injury. I-R injury impairs the healing at the intestinal anastomotic site. This study was designed to assess the role of aminoguanidine (AG, a selective inhibitor of iNOS), in healing at the colonic anastomotic site after intestinal I-R injury in rats.

Methods Female Wistar rats (n = 60) were divided into three groups. Group I (n = 15): sham operation, Group II (n = 15): I-R injury and anastomosis, and Group III (n = 30): I-R injury + anastomosis + AG-50 mg/kg. On the 7th postoperative day, relaparotomy was done and 4 cm of the colon with an intact area of the anastomosis was resected. Bursting pressure and histology at the anastomotic site were assessed.

Results The bursting pressure was significantly higher in Group III. In addition, bridging parameters (i.e., mucosal continuity, muscular continuity, re-epithelization, and granulation tissue), collagen pattern, and collagen density were significantly better in Group III. While the polymorphonuclear density was higher in Group II, suggestive of delayed healing.

Conclusion AG, by decreasing inflammation and increasing collagen content in an organized pattern, helped in preventing I-R injury at the site of colonic anastomosis in rats.

Keywords

ischemia reperfusion injury - nitric oxide synthase - aminoguanidine - intestinal anastomosis healing

• References

• 1 Cappell MS. Intestinal (mesenteric) vasculopathy. II. Ischemic colitis and chronic mesenteric ischemia. Gastroenterol Clin North Am 1998; 27 (4) 827-860 , vi
• 2 Homer-Vanniasinkam S, Crinnion JN, Gough MJ. Post-ischaemic organ dysfunction: a review. Eur J Vasc Endovasc Surg 1997; 14 (3) 195-203
• 3 Mitsudo S, Brandt LJ. Pathology of intestinal ischemia. Surg Clin North Am 1992; 72 (1) 43-63
• 4 Fielding LP, Stewart-Brown S, Blesovsky L, Kearney G. Anastomotic integrity after operations for large-bowel cancer: a multicentre study. BMJ 1980; 281 (6237) 411-414
• 5 Thornton FJ, Barbul A. Healing in the gastrointestinal tract. Surg Clin North Am 1997; 77 (3) 549-573
• 6 McMillen MA, Huribal M, Sumpio B. Common pathway of endothelial-leukocyte interaction in shock, ischemia, and reperfusion. Am J Surg 1993; 166 (5) 557-562
• 7 Kologlu M, Yorganci K, Renda N, Sayek I. Effect of local and remote ischemia-reperfusion injury on healing of colonic anastomoses. Surgery 2000; 128 (1) 99-104
• 8 Demiroğullari B, Sönmez K, Türkyilmaz Z , et al. Comparison of consequent small bowel anastomoses after transient ischemia: an experimental study in rats. J Pediatr Surg 1998; 33 (1) 91-93
• 9 Colak T, Turkmenoglu O, Dag A , et al. The effect of remote ischemic preconditioning on healing of colonic anastomoses. J Surg Res 2007; 143 (2) 200-205
• 10 Granger DN. Role of xanthine oxidase and granulocytes in ischemia-reperfusion injury. Am J Physiol 1988; 255 (6) (Pt 2) H1269-H1275
• 11 Nagira M, Tomita M, Mizuno S, Kumata M, Ayabe T, Hayashi M. Ischemia/reperfusion injury in the monolayers of human intestinal epithelial cell line caco-2 and its recovery by antioxidants. Drug Metab Pharmacokinet 2006; 21 (3) 230-237
• 12 Wasserberg N, Pileggi A, Salgar SK , et al. Heme oxygenase-1 upregulation protects against intestinal ischemia/reperfusion injury: a laboratory based study. Int J Surg 2007; 5 (4) 216-224
• 13 Fotiadis C, Adamis S, Misiakos EP , et al. The prophylactic effect of L-arginine in acute ischaemic colitis in a rat model of ischaemia/reperfusion injury. Acta Chir Belg 2007; 107 (2) 192-200
• 14 Xie Q, Nathan C. The high-output nitric oxide pathway: role and regulation. J Leukoc Biol 1994; 56 (5) 576-582
• 15 Suzuki Y, Deitch EA, Mishima S, Lu Q, Xu D. Inducible nitric oxide synthase gene knockout mice have increased resistance to gut injury and bacterial translocation after an intestinal ischemia-reperfusion injury. Crit Care Med 2000; 28 (11) 3692-3696
• 16 Vivar JV, Kalyanaraman B, Martásek P , et al. Superoxide generation by endothelial nitric oxide synthase: the influence of cofactors. Proc Natl Acad Sci U S A 1998; 95 (16) 9220-9225
• 17 Mungrue IN, Husain M, Stewart DJ. The role of NOS in heart failure: lessons from murine genetic models. Heart Fail Rev 2002; 7 (4) 407-422
• 18 Corbett JA, McDaniel ML. The use of Aminoguanidine: a selective iNOS inhibitor, to evaluate the role of nitric oxide in the development of autoimmune diabetes. Methods 1996; 10 (1) 21-30
• 19 Misko TP, Moore WM, Kasten TP , et al. Selective inhibition of the inducible nitric oxide synthase by aminoguanidine. Eur J Pharmacol 1993; 233 (1) 119-125
• 20 Kabali B, Girgin S, Gedik E, Ozturk H, Kale E, Buyukbayram H. N-acetylcysteine prevents deleterious effects of ischemia/reperfusion injury on healing of colonic anastomosis in rats. Eur Surg Res 2009; 43 (1) 8-12
• 21 Parks DA, Granger DN. ontributions of ischemia and reperfusion to mucosal lesion formation. Am J Physiol 1986; 250 (6) (Pt 1) G749-G753
• 22 Neary P, Redmond HP. Ischaemia-reperfusion injury and the systemic inflammatory response syndrome. In: Grace PA, Mathie RT, , eds. Ischaemia-Reperfusion Injury. London: Blackwell Science; 1999: 123-126
• 23 Kuzu MA, Tanik A, Kale IT, Aşlar AK, Köksoy C, Terzi C. Effect of ischemia/reperfusion as a systemic phenomenon on anastomotic healing in the left colon. World J Surg 2000; 24 (8) 990-994
• 24 Erbil Y, Calis A, Berber E, Mercan S. The effect of intraoperative colonic lavage with NG-nitro-L-arginine methyl ester (L-NAME) on anastomotic healing in the presence of left-sided colonic obstruction in the rat. Surg Today 2000; 30 (5) 421-425
• 25 Polat C, Arikan Y, Gokce C , et al. The effect of NG-nitro L-arginine methyl ester on colonic anastomosis after increased intra-abdominal pressure. Langenbecks Arch Surg 2007; 392 (2) 197-202
• 26 Wise L, McAlister W, Stein T, Schuck P. Studies on the healing of anastomoses of small and large intestines. Surg Gynecol Obstet 1975; 141 (2) 190-194
• 27 Schiller HJ, Reilly PM, Bulkley GB. Tissue perfusion in critical illnesses. Antioxidant therapy. Crit Care Med 1993; 21 (2 Suppl) S92-S102
• 28 Kuzu MA, Köksoy C, Kale IT, Tanik A, Terzi C, Elhan AH. Reperfusion injury delays healing of intestinal anastomosis in a rat. Am J Surg 1998; 176 (4) 348-351
• 29 Rashid G, Luzon AA, Korzets Z, Klein O, Zeltzer E, Bernheim J. The effect of advanced glycation end-products and aminoguanidine on TNFalpha production by rat peritoneal macrophages. Perit Dial Int 2001; 21 (2) 122-129
• 30 Ara C, Karabulut AB, Kirimlioglu H, Yilmaz M, Kirimliglu V, Yilmaz S. Protective effect of aminoguanidine against oxidative stress in an experimental peritoneal adhesion model in rats. Cell Biochem Funct 2006; 24 (5) 443-448
• 31 Szabó C, Southan GJ, Thiemermann C. Beneficial effects and improved survival in rodent models of septic shock with S-methylisothiourea sulfate, a potent and selective inhibitor of inducible nitric oxide synthase. Proc Natl Acad Sci U S A 1994; 91 (26) 12472-12476
• 32 Garcin ED, Arvai AS, Rosenfeld RJ , et al. Anchored plasticity opens doors for selective inhibitor design in nitric oxide synthase. Nat Chem Biol 2008; 4 (11) 700-707
• 33 Cohen IK, Robert F, Diegelman S. Wound care and wound healing. In: Schwartz SI, Shires TG, Spencer CF, , eds. Principles of Surgery. 7th ed. New York, NY: McGraw-Hill; 1999: 263-295
• 34 Hacıyanlı M, Fuzun M, Ünek T, Tokgoz Z. Does the administration route of leucovorin have any influence on the impairment of colonic healing caused by intraperitoneal 5-fluorouracil treatment?. Eur Surg Res 2001; 33 (2) 80-85