Local Administration of Adenosine and Adenosine A1 Receptor Agonist CPA Protects Against Intestinal Ischemia-Reperfusion Injury in Rats

Abstract

Objective: Adenosine and adenosine A1 receptor (A1AR) agonists have potential protective effects against reperfusion injury in variety of tissues. The purpose of the present study was to investigate possible effects of topical administration of adenosine and A1AR agonist on reperfusion-induced small intestinal injury in rat. Materials and Methods: Rats were randomized to five groups each including six as following: sham-operated control; ischemia-reperfusion (I/R) control; adenosine + I/R; A1AR agonist 2-chloro-N6-cyclopentyladenosine (CPA) + I/R; and A1AR antagonist, 8-cyclopentyl-1,3-dipropylxanthine (DPCPX) + adenosine + I/R. Following topical administration of the drugs into abdominal cavity for 5 min, intestinal I/R was established by clamping superior mesenteric artery (SMA) for 30 minutes followed by 180 min of reperfusion period. Afterwards terminal ileum samples were collected and immediately transferred to isolated organ bath for measuring contractile response to carbachol. Furthermore, additional tissue samples were harvested for measuring the levels of malondialdehyde (MDA) and reduced glutathione (GSH). Results: I/R significantly increased lipid peroxidation while decreasing the GSH. Contractile responses were seriously reduced in I/R group compared to that of the sham control group. Pretreatment with adenosine or CPA not only decreased lipid peroxidation but also ameliorated contractile response and GSH levels remarkably. These beneficial effects were abolished by pretreatment with A1AR antagonist DPCPX. Conclusion: Evidences we collected suggest that besides systemic administration, local application of adenosine and A1AR agonist CPA also attenuate ischemic intestinal injury via, at least, decreasing oxidative stress and enhancing antioxidant defense.

Keywords

• -

Adenozin ve Adenozin A1 Reseptör Agonisti CPA'nın Lokal Uygulanımı Sıçan İnce Barsak Hasarına Karsı Koruyucu Etkiler Göstermektedir

Öz

Amaç: Adenozin ve A1 adenozin reseptör (A1AR) agonistleri, çeşitli dokuların reperfüzyon hasarına karşı koruyucu etkilere sahiptir. Çalışmanın amacı, sıçan ince barsağının reperfüzyon harabiyetinde adenozin ve A1AR agonistinin lokal uygulanmasının etkilerini incelemekti. Gereç ve Yöntemler: Sıçanlar herbiri altı hayvan içeren beş gruba rastgele olarak aşağıdaki gibi ayrıldı: sham kontrol; iskemi-reperfüzyon (I/R) kontrol; adenozin + I/R; A1AR agonisti 2-kloro-N6-siklopentiladenozin (CPA) + I/R ve A1AR antagonisti 8-siklopentil-1,3-dipropilksantin (DPCPX) + adenozin + I/R. Đlaçların abdominal boşluğa 5 dk boyunca lokal uygulanmasını takiben barsak I/R’u, süperiyor mezenter arterin 30 dk klempe edilmesi ve sonrasındaki 180 dk lık reperfüzyon dönemi ile sağlandı. Daha sonra terminal ileum örnekleri toplandı ve karbakole olan kasılma yanıtlarını ölçmek için hızlıca izole organ banyosuna alındı. Ayrıca malondialdehid (MDA) ve indirgenmiş glutatyon (GSH) düzeylerini ölçmek için de ek doku örnekleri alındı. Bulgular: I/R, lipid peroksidasyonunu ileri düzeyde yükseltirken, indirgenmiş glutatyonu düşürdü. Sham kontrol grubuyla karşılaştırıldığında, kasılma yanıtları I/R grubunda ciddi düzeyde azaldı. Adenozin veya CPA ön tedavisi, sadece lipit peroksidasyonunu azaltmakla kalmadı aynı zamanda kasılma yanıtı ve GSH düzeyini de ileri derecede iyileştirdi. Bu yararlı etkilerin, A1AR anatgonisti DPCPX ön tedavisi ile ortadan kalktığı gözlendi. Sonuç: Elde ettiğimiz kanıtlar, sistemik uygulanımının yanı sıra, adenozin ve A1AR agonisti CPA’nın lokal uygulanmasının da iskemik barsak harabiyetini, en azından oksidatif stresi azaltarak ve antioksidan savunmayı güçlendirerek iyileştirdiğini önermektedir.

Anahtar Kelimeler

• Adenozin, adenozin A1 reseptörü, barsak iskemi/reperfüzyonu

References

1. Mallick IH, Yang W, Winslet MC, Seifaian AM. Ischemiareperfusion injury of the intestine and protective strategies against injury. Dig Dis Sci 2004; 49:1359-1377.
2. Lodato RF, Khan RA, Zembowicz MJ, et al. Roles of IL-1 and TNF in the decreased ileal muscle contractility induced by lipopolysaccaride. Am J Physiol 1999; 276:1356-1362.
3. Carden DL ve Granger DN. Pathophysiology of ischemiareperfusion injury. J Pathol 2000; 190: 255-266.
4. Arumugam TV, Shiels IA, Woodruff TM, et al. Protective effect of a new C5a receptor antagonist against ischemireperfusion injury in the rat small intestine. J Surg Res 2002; 103:260-267.
5. Bielefeldt K ve Conklin JI. Intestinal motility during hypoxia and reoxygenation in vitro. Dig Dis Sci 1997; 42:878-884.
6. Khanna A, Rossman JE, Fung HL, Caty MG. Attenuated nitric oxide synthase activity and protein expression accompany intestinal ischemia/reperfusion injury in rats. Biochem Biophys Res Commun 2000; 269: 160-164.
7. Poussios D, Andreadou I, Papalois A, et al. Protective effect of a novel antioxidant non-steroidal anti-inflammatory agent (compound IA) on intestinal viability after acute mesenteric ischemia and reperfusion. Eur J Pharmacol 2003; 465: 275-280.
8. Takahashi A, Tomomasa T, Kaneko H et al. Intestinal motility in an in vitro rat model of intestinal ischemia-reperfusion with special reference to the effects of nitric oxide on the motility changes. J Pediatr Gastroenterol Nutr 2001; 33:283-288.

9. Ozacmak VH, Sayan H, Arslan SO, Altaner S, Aktas RG. Protective effect of melatonin on contractile activity and oxidative injury induced by ischemia and reperfusion of rat ileum. Life Sci 2005; 76: 1575-1588.
10. Hassoun HT, Weisbrodt NW, Mercer DW, et al. Inducible nitric oxide synathase mediates gut ischemia/reperfusioninduced ileus only after severe insults. J Surg Res 2001; 97: 150-154.
11. Baxter GF. Role of adenosine in delayed preconditioning of myocardium. Cardiovasc Res 2002; 55: 483-494.
12. Mubagwa K ve Flameng W. Adenosine, adenosine receptors and myocardial protection: an updated overview. Cardivasc Res 2001; 52:25-39.
13. Headrick JP, Hack B, Ashton KJ. Acute adenosinergic cardioprotection in ischemic-reperfused hearts. Am J Physiol Heart Circ Physiol 2003; 285:1797-1818.
14. Solenkova NV, Solodushko V, Cohen MV, Downey JM. Endogenous adenosine protects preconditioned heart during early minutes of reperfusion by activating Akt. Am J Physiol Heart Circ Physiol 2006; 290 (1):441-449.
15. Dana A, Jonassen AK, Yamashita N, Yellon DM. Adenosine A1 receptor activation induces delayed preconditioning in rats mediated by manganese superoxide dismutase. Circulation 2000; 101:2841-2848.
16. Kristo G, Yoshimura Y, Keith BJ, et al. Adenosine A1/A2a receptor agonist AMP-579 induces acute and delayed preconditioning against in vivo myocardial stunning. Am J Physiol Heart Circ Physiol 2004; 287:2746-2753.
17. Mozzicato S, Joshi BV, Jacobsen KA, Liang BT. Role of direct RhoA-phospholipase D interaction in mediating adenosine-induced protection from cardiac ischemia. FASEB J 2004; 18(2):406-408.
18. De Jonge R, Out M, Maas WJ, De Jong JW. Preconditioning of rat hearts by adenosine A1 and A3 receptor activation. Eur J Pharmacol 2002; 441:165-172.
19. Maddock HL, Mocanu MM, Yellon DM. Adenosine A3 receptor activation protects the myocardium from reperfusion/reoxygenation injury. Am J Physiol Heart Circ Physiol 2002; 283:1307-1313.
20. Lee HT, Gallos G, Nasr SH, Emala CW. A1 adenosine receptor activation inhibits inflammation, necrosis, and apoptosis after renal ischemia-reperfusion injury in mice. J Am Soc Nephrol 2004; 15:102-111.
21. Randhawa MP, Lasley RD, Mentzer RM. Salutary effects of exogenous adenosine administration on in vivo myocardial stunning. J Thorac Cardiovasc Surg 1995; 110:63-74.
22. Sekili S, Jeroudi MO, Tang XL, et al. Effect of adenosine on myocardial stunning in the dog. Circ Res 1995; 76:82-94.
23. Yoshimura Y, Kristo G, Keith BJ ve ark. The p38 MAPK inhibitor SB203580 blocks adenosine A1 receptor-induced attenuation of in vivo myocardial stunning. Cardiovasc Drugs and Ther 2004; 18: 433-440.
24. Sugino H, Shimada H, Tsucimoto K. Role of adenosine in renal protection induced by a brief episode of ischemic preconditioning. Jpn J Pharmacol 2001; 87:134-142.
25. Nakamura M, Nakakimura K, Matsumoto M, Sakabe T. Rapid tolerance to focal cerebral ischemia in rats is attenuated by adenosine A1 receptor antagonist. J Cereb Blood Flow Metab 2002; 22(2):161-170.
26. Hiraide T, Katsura K, Muramatsu H, Asano G, Katayama Y. Adenosine receptor antagonists cancelled the ischemic tolerance phenomenon in gerbil. Brain Res 2001; 910(1-2):94-98.
27. Kudo M, Wang Y, Xu M, Ayup A, Ashraf M. Adenosine A1 receptor mediates late preconditioning via activation of PKCdelta signaling pathway. Am J Physiol Heart Circ Physiol 2002; 283: 296-301.
28. Zhao X, Alexander JS, Zhang S, et al. Redox regulation of endothelial barrier integrity. Am J Physiol Lung Cell Mol Physiol 2001; 281: 879-886.
29. Reid EA, Kristo G, Yoshimura Y, et al. In vivo adenosine receptor preconditioning reduces myocardial infarct size via subcellular ERK signaling. Am J Physiol Heart Circ Physiol 2005; 288(5): 2253-2259.
30. Ballard-Croft C, Kristo G, Yoshimura Y, et al. Acute adenosine preconditioning is mediated by p38 MAPK activation in discrete subcellular compartments. Am J Physiol Heart Circ Physiol 2005; 288: 1359-1366.
31. Ramkumar V, Hallam DM, Nie Z. Adenosine, oxidative stress and cytoprotection. Jpn J Pharmacol 2001; 86:265-274.
32. Kadowaki M, Tokita K, Nagakura Y, et al. Adenosine A1 receptor blockade reverses dysmotility induced by ischemiareperfusion in rat colon. Eur J Pharmacol 2000; 409:319-323.
33. Nicholls J ve Hourani SM. Characterization of adenosine receptors on rat ileum, ileal longitudinal muscle and muscularis mucosae. Eur J Pharmacol 1997; 338:143-150.
34. Ozacmak VH ve Sayan H. Pretreatment with adenosine and adenosine A1 receptor agonist protects against intestinal ischemia-reperfusion injury in rat. World J Gastroenterol 2007; 13(4):538-547.
35. Boucher M, Wann PB, Kaloustian S, et al. Sustained cardioprotection afforded by A2A adenosine receptor stimulation after 72 hours of myocardial reperfusion. J Cardiovasc Pharmacol 2005; 45(5):439-446.
36. Boucher M, Pesant S, Falcao S, et al.. Post-ischemic cardioprotection by A2A adenosine receptors: Dependent of phosphatidylinositol 3- kinase pathway. J Cardiovasc Pharmacol 2004; 43(3):416-422.
37. Davis JM, Gute DC, Jones S, Krsmanovic A, Korthius RJ. Ischemic preconditioning prevents postischemic P-selectin expression in the rat small intestine. Am J Physiol Heart Circ Physiol 1999; 46: 2476-2481.
38. Unal S, Demirkan F, Arslan E, et al. Comparison of ischemic and chemical preconditioning in jejunal flaps in the rat. Plast Reconstr Surg 2003; 112:1024-1031.
39. Casini A, Ferrali M, Pampella A, Maellaro E, Comporti M. Lipid peroxidation and cellular damage in extrahepatic tissues of bromobenzene-intoxicated mice. Am J Pathol 1986; 123: 520-531.
40. Aykac G, Uysal M, Yalan AS, et al. The effects of chronic ethanol injection on hepatic lipid peroxide, glutathione, glutathione peroxidase and glutathione transferase in rats. Toxicology 1985; 36:71-76.
41. Ferrer JV, Ariceta J, Guerrero D, et al. Allopurinol and Nacetylcysteine avoid 60% of intestinal necrosis in an ischemiareperfusion experimental model. Transplant Proc 1998; 30(6): 2672.
42. Jacob T, Ascher E, Hingorani A, Kallakuri S. Glycine prevents the induction of apoptosis attributed to mesenteric ischemiareperfusion injury in a rat model. Surgery 2003; 134(3): 457466.
43. Lee HT, Xu H, Nasr SH, Schnermann J, Emala CW. A1 adenosine receptor knockout mice exhibit increased renal injury following ischemia-reperfusion. Am J Physiol Renal Physiol 2004; 286:298-306.
44. Lee HT ve Emala CW. Adenosine attenuates oxidant injury in human proximal tubular cells via A (1) and A (2a) adenosine receptors. Am J Physiol Renal Physiol 2002; 282: 844-852.
45. Narayan P, Mentzer RM, Lasley RD. Adenosine A1 receptor activation reduces reactive oxygen species and attenuates stunning in ventricular myocytes. J Mol Cell Cardiol 2001; 33: 221-129.
46. Thomas GP, Sims SM, Cook MA, Karmazyn M. Hydrogen peroxide-induced stimulation of L-type calcium current in guinea pig ventricular myocytes and its inhibition by adenosine A1 receptor activation. J Pharmacol Exp Ther 1998; 286:1208-1214.