Acute pancreatitis (AP) is a common inflammatory disease mediated by damage to acinar cells and subsequent pancreatic inflammation with infiltration of leukocytes. The neuronal guidance protein, netrin-1, has been shown to control leukocyte trafficking and modulate inflammatory responses in several inflammation-based diseases. The present study was aimed toward investigating the effects of netrin-1 in an in vivo model of AP in mice. AP was induced in C57BL/6 mice by administration of two intraperitoneal injections of L-Arginine (4 g/kg). Mice were treated with recombinant mouse netrin-1 at a dose of 1 μg/mouse or vehicle (0.1% BSA) intravenously through the tail vein immediately after the second injection of L-Arginine, and every 24 h thereafter. Mice were sacrificed at several time intervals from 0 to 96 h after the induction of pancreatitis. Blood and tissue samples of pancreas and lung were collected and processed to determine the severity of pancreatitis biochemically and histologically. Immunohistochemical staining demonstrated that netrin-1 was mainly expressed in the islet cells of the normal pancreas and the AP model pancreas, and the pancreatic expression of netrin-1 was down-regulated at both the mRNA and protein levels during the course of AP. Exogenous netrin-1 administration significantly reduced plasma amylase levels, myeloperoxidase activity, pro-inflammatory cytokine production, and pancreas and lung tissue damages. Furthermore, netrin-1 administration did not cause significant inhibition of nuclear factor-kappa B activation in the pancreas of L-Arginine-induced AP. In conclusion, our novel data suggest that netrin-1 is capable of improving damage of pancreas and lung, and exerting anti-inflammatory effects in mice with severe acute pancreatitis. Thus, our results indicate that netrin-1 may constitute a novel target in the management of AP.
References
[1]
Bhatia M, Wong FL, Cao Y, Lau HY, Huang JL, et al. (2005) Pathophysiology of acute pancreatitis. Pancreatology 5: 134–144.
[2]
Gravante G, Garcea G, Ong SL, Metcalfe MS, Berry DP, et al. (2009) Prediction of mortality in acute pancreatitis: a systematic review of the published evidence. Pancreatology 9: 601–614.
[3]
Whitcomb DC (2006) Clinical practice. Acute pancreatitis. N Engl J Med 354: 2142–2150.
[4]
Pezzilli R (2009) Pharmacotherapy for acute pancreatitis. Expert Opin Pharmacother 10: 2999–3014.
[5]
Vonlaufen A, Wilson JS, Apte MV (2008) Molecular mechanisms of pancreatitis: current opinion. J Gastroenterol Hepatol 23: 1339–1348.
[6]
Serafini T, Kennedy TE, Galko MJ, Mirzayan C, Jessell TM, et al. (1994) The netrins define a family of axon outgrowth-promoting proteins homologous to C. elegans UNC-6. Cell 78: 409–424.
[7]
Kennedy TE, Serafini T, de la Torre JR, Tessier-Lavigne M (1994) Netrins are diffusible chemotropic factors for commissural axons in the embryonic spinal cord. Cell 78: 425–435.
[8]
Cirulli V, Yebra M (2007) Netrins: beyond the brain. Nat Rev Mol Cell Biol 8: 296–306.
[9]
Ly NP, Komatsuzaki K, Fraser IP, Tseng AA, Prodhan P, et al. (2005) Netrin-1 inhibits leukocyte migration in vitro and in vivo. Proc Natl Acad Sci USA 102: 14729–14734.
[10]
Wang W, Reeves WB, Ramesh G (2008) Netrin-1 and kidney injury. I. Netrin-1 protects against ischemia-reperfusion injury of the kidney. Am J Physiol Renal Physiol 294: F739–F747.
[11]
Rosenberger P, Schwab JM, Mirakaj V, Masekowsky E, Mager A, et al. (2009) Hypoxia-inducible factor-dependent induction of netrin-1 dampens inflammation caused by hypoxia. Nat Immunol 10: 195–202.
[12]
Mirakaj V, Thix CA, Laucher S, Mielke C, Morote-Garcia JC, et al. (2010) Netrin-1 dampens pulmonary inflammation during acute lung injury. Am J Respir Crit Care Med 181: 815–824.
Aherne CM, Collins CB, Masterson JC, Tizzano M, Boyle TA, et al. (2011) Neuronal guidance molecule netrin-1 attenuates inflammatory cell trafficking during acute experimental colitis. Gut 10: 1136–1143.
[15]
De Breuck S, Lardon J, Rooman I, Bouwens L (2003) Netrin-1 expression in fetal and regenerating rat pancreas and its effect on the migration of human pancreatic duct and porcine islet precursor cells. Diabetologia 46: 926–933.
[16]
Dumartin L, Quemener C, Laklai H, Herbert J, Bicknell R, et al. (2010) Netrin-1 mediates early events in pancreatic adenocarcinoma progression, acting on tumor and endothelial cells. Gastroenterology 138: 1595–1606.
[17]
Yang YHC, Szabat M, Bragagnin C, Kott K, Helgason CD, et al. (2011) Paracrine signalling loops in adult human and mouse pancreatic islets: netrins modulate beta cell apoptosis signalling via dependence receptors. Diabetologia 54: 828–842.
[18]
Dawra R, Sharif R, Phillips P, Dudeja V, Dhaulakhandi D, et al. (2007) Development of a new mouse model of acute pancreatitis induced by administration of L-arginine. Am J Physiol Gastrointest Liver Physiol 292: G1009–1018.
[19]
Zhou XL, Xue CR (2009) Ghrelin inhibits the development of acute pancreatitis and nuclear factor kappaB activation in pancreas and liver. Pancreas 38: 752–757.
[20]
Sandoval D, Gukovskaya A, Reavey P, Gukovsky S, Sisk A, et al. (1996) The role of neutrophils and platelet-activating factor in mediating experimental pancreatitis. Gastroenterology 111: 1081–1091.
[21]
Werner J, Dragotakes SC, Fernandez-del Castillo C, Rivera JA, Ou J, et al. (1998) Technetium-99m-labeled white blood cells: a new method to define the local and systemic role of leukocytes in acute experimental pancreatitis. Ann Surg 227: 86–94.
[22]
Bhatia M, Saluja AK, Hofbauer B, Lee HS, Frossard JL, et al. (1998) The effects of neutrophil depletion on a completely noninvasive model of acute pancreatitis-associated lung injury. Int J Pancreatol 24: 77–83.
[23]
Inoue S, Nakao A, Kishimoto W, Murakami H, Itoh K, et al. (1995) Anti-neutrophil antibody attenuates the severity of acute lung injury in rats with experimental acute pancreatitis. Arch Surg 130: 93–98.
[24]
Norman J (1998) The role of cytokines in the pathogenesis of acute pancreatitis. Am J Surg 175: 76–83.
[25]
Pereda J, Sabater L, Aparisi L, Escobar J, Sandoval J, et al. (2006) Interaction between cytokines and oxidative stress in acute pancreatitis. Curr Med Chem 13: 2775–2787.
[26]
Norman J, Franz M, Messina J, Riker A, Fabri PJ, et al. (1995) Interleukin-1 receptor antagonist decreases severity of experimental acute pancreatitis. Surgery 117: 648–655.
[27]
Hughes CB, Grewal HP, Gaber LW, Kotb M, El-din AB, et al. (1996) Anti-TNFalpha therapy improves survival and ameliorates the pathophysiologic sequelae in acute pancreatitis in the rat. Am J Surg 171: 274–280.
[28]
Javier P, Luis S, Norberto C, Luis GC, Daniel C, et al. (2004) Effect of simultaneous inhibition of TNF-a production and xanthine oxidase in experimental acute pancreatitis. Ann Surg 240: 108–116.
[29]
Chen X, Ji B, Han B, Ernst SA, Simeone D, et al. (2002) NF-kappaB activation in pancreas induces pancreatic and systemic inflammatory response. Gastroenterology 122: 448–457.
[30]
Rakonczay Z Jr, Ja'rmay K, Kaszaki J, Mándi Y, Duda E, et al. (2003) NF-kappaB activation is detrimental in arginine-induced acute pancreatitis. Free Radic Biol Med 34: 696–709.
[31]
Ethridge RT, Hashimoto K, Chung DH, Ehlers RA, Rajaraman S, et al. (2002) Selective inhibition of NF-kappaB attenuates the severity of cerulein-induced acute pancreatitis. J Am Coll Surg 195: 497–505.
[32]
Satoh A, Shimosegawa T, Fujita M, Kimura K, Masamune A, et al. (1999) Inhibition of nuclear factor-kappaB activation improves the survival of rats with taurocholate pancreatitis. Gut 44: 253–258.
[33]
Steinle AU, Weidenbach H, Wagner M, Adler G, Schmid RM (1999) NF-kappaB/Rel activation in cerulein pancreatitis. Gastroenterology 116: 420–430.
[34]
Algül H, Treiber M, Lesina M, Nakhai H, Saur D, et al. (2007) Pancreas-specific RelA/p65 truncation increases susceptibility of acini to inflammation-associated cell death following cerulein pancreatitis. J Clin Invest 117: 1490–1501.
[35]
Chen H, Sun YP, Li Y, Yan RL, Liu WW, et al (2010) Hydrogen-rich saline ameliorates the severity of l-arginine-induced acute pancreatitis in rats. Biochem Biophys Res Commun 393 (2) 308–313.
[36]
Paradisi A, Maisse C, Bernet A, Coissieux M, Maccarrone M, et al. (2008) NF-κB regulates netrin-1 expression and affects the conditional tumor suppressive activity of the netrin-1 receptors. Gastroenterology 135: 1248–1257.
[37]
Renard P, Ernest I, Houbion A, Art M, Le Calvez H, et al. (2001) Development of a sensitive multi-well colorimetric assay for active NFkB. Nucleic Acids Res 29 (4) E21.
[38]
Mukhopadhyay A, Bueso-Ramos C, Chatterjee D, Pantazis P, Aggarwal BB (2001) Curcumin downregulates cell survival mechanisms in human prostate cancer cell lines. Oncogene 20: 7597–7609.
[39]
Tsuchiya A, Hayashi T, Deguchi K, Sehara Y, Yamashita T, et al. (2007) Expression of netrin-1 and its receptors DCC and neogenin in rat brain after ischemia. Brain Res 1159: 1–7.
[40]
Llambi F, Causeret F, Bloch-Gallego E, Mehlen P (2001) Netrin-1 acts as a survival factor via its receptors UNC5H and DCC. EMBO J 20: 2715–2722.
[41]
Tang XL, Jang SW, Okada M, Chan CB, Feng Y, et al. (2008) Netrin-1 mediates neuronal survival through PIKE-L interaction with the dependence receptor UNC5B. Nat Cell Biol 10: 698–706.
[42]
Wang W, Reeves WB, Ramesh G (2009) Netrin-1 increases proliferation and migration of renal proximal tubular epithelial cells via the UNC5B receptor. Am J Physiol Renal Physiol 296: F723–F729.
[43]
Tadagavadi RK, Wang W, Ramesh G (2010) Netrin-1 regulates Th1/Th2/Th17 cytokine production and inflammation through UNC5B receptor and protects kidney against ischemia-reperfusion injury. J Immunol 185: 3750–3758.
[44]
Paradisi A, Maisse C, Coissieux MM, Gadot N, Lépinasse F, et al. (2009) Netrin-1 up-regulation in inflammatory bowel diseases is required for colorectal cancer progression. Proc Natl Acad Sci USA 106: 17146–17151.
[45]
Link BC, Reichelt U, Schreiber M, Kaifi JT, Wachowiak R, et al. (2007) Prognostic implications of netrin-1 expression and its receptors in patients with adenocarcinoma of the pancreas. Ann Surg Oncol 14: 2591–2599.
