Protective effect of sulfasalazine on the intestinal ischemia-reperfusion injury

Ramazan Eryılmaz; Mustafa Şahin; Tamer Gündoğdu  Adem Akçakaya; Orhan Alimoğlu; İsmail Okan; Adnan Somay; M. Baki Çekmen; Emin Özbek

Abstract

Purpose: Acute mesenteric occlusion is a serious clinical entity which usually requires prompt surgical intervention. Although reperfusion is mandatory for the repair, the release of free oxygen radicals and the induction of neutrophyl accumulation attenuates the injury. NFkappa B has a critical role in inflammatory process. This study is based on the hypothesis that sulfasalazine, a potent inhibitor of NF-kappa B may counteract or decrease the ischemia/reperfusion (I/R) injury in rats. Materials and Methods: The study consisted of three groups each containing 10 rats. In the control group (Group I) superior mesenteric artery was dissected, but not ligated. Two hours later tissue samples from ileum and blood samples from portal vein were taken and the rats were sacrificed. In I/R group (Group II) after dissection of superior mesenteric artery, it was clamped just distal to the origin from aorta. Clamp is released after 1 hour ischemia and tissues were reperfused. The same samples as in control group were taken. In sulfasalazin group (Group III), apart from group II, 30 minutes after the beginning of the ischemia, sulfasalazine was given. The tissue samples from ileum were stained for iNOS, p65 and caspase immunohistochemically. NO level was determined in blood samples by Greiss reaction. iNOS, p65 and caspase protein expressions in tissue sections and NO levels in serum were at the basal level in control group. Results: In I/R group both protein expressions and serum NO levels were very high. However in sulfasalazine group, these parameter levels were found higher than control group, but less than the levels found in I/R group. Statistical analysis showed that although there was significant difference between control group and I/R group (p<0.05) with regard to serum NO levels, iNOS and caspase expressions, sulfasalazin group has no significant difference when compared with control group (p>0.05). p65 expression was found to be significantly increased in I/R group compared to control group (p<0.05), whereas in sulfasalazine given group the expression decreased compared to I/R group. However, the decrement was not statistically significant. Conclusion: This study demonstrated that sulfasalazin may have protective effect on experimental rat intestinal ischemia-reperfusion injury through down-regulation of tissue proteins responsible from celluler damage.

Keywords:

Intestine, ischemia-reperfusion injury, sulfasalazine

References

Boley SJ, Brandt LJ, Sammartano RJ. History of mesenteric ischemia. The evolution of a diagnosis and management. Surg Clin North Am 1997;77:275-288.

Mitchell RN, Cotran RS. Cellular pathology I: Cell injury and cell death. In: Robbins Pathologic Basis of Disease. 6th edition. Philadelphia: WB Saunders Company. 1999:5-28.

Grace PA. Ischaemia-reperfusion injury. Br J Surg 1994;81:637-647.

Lin E, Calvano SE, Lowry SF. Systemic response to injury and metabolic support. In: Brunicardi FC, Andersen DK, Billian TR, Dunn DL, Hunter JG, Pollock RE, eds. Schwartz\'s Principles of Surgery. 8th edition. New York: McGraw-Hill Medical Publishing Division. 2005:1-32.

Girotti AW. Lipid hydroperoxide generation, turnover, and effector action in biological systems. J Lipid Res 1998;39:1529-1542.

Lille ST, Lefler SR, Mowlavi A, et al. Inhibition of the initial wave of NF-kappaB activity in rat muscle reduces ischemia/reperfusion injury Muscle Nerve 2001;24:534-541.

Sasaki H, Zhu L, Fukuda S, Maulik N. Inhibition of NF-kappaB activation by pyrrolidine dithiocarbamate prevents in vivo hypoxia/reoxygenationmediated myocardial angiogenesis. Int J Tissue React 2000;22:93-100.

Wu B, Iwakiri R, Tsunada S, et al. iNOS enhances rat intestinal apoptosis after ischemia-reperfusion. Free Radic Biol Med 2002;33:649-658.

Zimmerman BJ, Granger DN. Mechanisms of reperfusion injury. Am J Med Sci 1994;307;284-

Akcakaya A, Alimoglu O, Sahin M, Abbasoglu SD. Ischemia-reperfusion injury following superior mesenteric artery occlusion and strangulation obstruction. J Surg Res 2002;108:39-43.

Kalia N, Pockley AG, Wood RF, Brown NJ. Effect of FK409 on intestinal ischemia-reperfusion injury and ischemia induced changes in the rat mucosal villus microcirculation. Transplantation 2001;72:1875-1880.

Ross SD, Kron IL, Gangemi JJ et al. Attenuation of lung reperfusion injury after transplantation using an inhibitor of nuclear factor- B. Am J Physiol Lung Cell Mol Physiol 2000;279:L528-536.

Abraham E. NF-kappa B activation. Crit Care Med 2000;28:100-104.

Shen WH, Zhang CY, Zhang GY. Antioxidants attenuate reperfusion injury after global brain ischemia through inhibiting nuclear factorkappa B activity in rats. Acta Pharmacol Sin 2003;24:1125-1130.

Khanna A, Rossman JE, Fung HL, Caty MG. Attenuated Nitric oxide synthase and protein expression accompany intestinal ischemia/reperfusion injury in rats. Biochem Biophys Res Commun 2000;269:160-164.

Yadav S, Sindram D, Perry DK, Clavien PA. Ischemic preconditioning protects the mouse liver by inhibition of apoptosis through a caspasedependant pathway. Hepatology 1999;30:1223-1231.

Liang F, Gao E, Tao L, et al. Critical timing of larginine treatment in post ischemic myocardial apoptosis-role of NOS isoforms. Cardiovasc Res 2004;62:568-577.

Iwakiri R, Gotoh Y, Noda T, et al. Programmed cell death in rat intestine: effect of feeding and fasting. Scand J Gastroenterol 2001;36:39-47.

Fujimoto K, Hosotani R, Wada M, et al. Ischemiareperfusion injury on the pancreas in rats: identification of acinar cell apoptosis. J Surg Res 1997;71:127-136.

Naito Y, Takagi T, Ichikawa H., et al. A novel potent inhibitor of inducible nitric oxide inhibitor, ONO-1714, reduces intestinal ischemia-reperfusion injury in rats. Nitric Oxide 2004;10:170-177.

Basoglu M, Yildirgan I, Akcay F, Kiziltunc A, Kava I, Oren D. Glutathione and nitric oxide concentrations in glutamine-infused rabbits with intestinal ischaemia/reperfusion. Eur J Clin Chem Clin Biochem 1997;35:415-419.

Cuzzocrea S, Zingarelli B, Caputi AP. Role of constitutive nitric oxide synthase and peroxinitrite production in a rat model of splanchnic artery occlusion shock. Life Sci 1998;63:789-799.

Chan KL, Zhang XH, Fung PC, Guo WH, Tam PK. Role of nitric oxide in intestinal ischaemia-reperfusion injury studied using electron paramagnetic resonance. Br J Surg 1999;86:1427-1432.

Nakashima O, Terada Y, Inoshita S, Kuwahara M, Sasaki S, Marumo F. Inducible nitric oxide synthase can be induced in the absence of active nuclear factor-kappaB in rat mesengial cells: involvement of the Janus kinase signalling pathway. J Am Soc Nephrol 1999;10:721-729.

Wahl C, Liptay S, Adler G, Schmid RM. Sulfasalazine: a potent and specific inhibitor of Nuclear factor kappa B. J Clin Invest 1998;101:1163-1174.

MacDermott RP. Progress in understanding the mechanism of action of 5-aminosalisylic acid. Am J Gastroenterol 2000;95:3343-3348.

Feeley BT, Park AK, Hoyt EG, Robbins RC. Sulfasalazine inhibits reperfusion injury and prolongs allograft survival in rat cardiac transplants J Heart Lung Transplant 1999;18:1088-1095.