EXPERIMENTAL RESEARCH
An optimised mouse model of chronic pancreatitis with a combination of ethanol and cerulein
Submission date: 2015-07-31
Final revision date: 2015-08-30
Acceptance date: 2015-09-07
Publication date: 2016-03-24
Cent Eur J Immunol 2016;41(1):54-63
KEYWORDS
ABSTRACT
Introduction: Chronic pancreatitis (CP) is an intractable and multi-factorial disorder. Developing appropriate animal models is an essential step in pancreatitis research, and the best ones are those which mimic the human disorder both aetiologically and pathophysiologically. The current study presents an optimised protocol for creating a murine model of CP, which mimics the initial steps of chronic pancreatitis in alcohol chronic pancreatitis and compares it with two other mouse models treated with cerulein or ethanol alone.
Material and methods: Thirty-two male C57BL/6 mice were randomly selected, divided into four groups, and treated intraperitoneally with saline (10 ml/kg, control group), ethanol (3 g/kg; 30% v/v), cerulein (50 µg/kg), or ethanol + cerulein, for six weeks. Histopathological and immunohistochemical assays for chronic pancreatitis index along with real-time PCR assessments for mRNA levels of inflammatory cytokines and fibrogenic markers were conducted to verify the CP induction.
Results: The results indicated that CP index (CPI) was significantly increased in ethanol-cerulein mice compared to the saline, ethanol, and cerulein groups (p < 0.001). Interleukin 1 (IL-1), tumor necrosis factor (TNF-), transforming growth factor (TGF-), -smooth muscle actin (-SMA), and myeloperoxidase activity were also significantly greater in both cerulein and ethanol-cerulein groups than in the saline treated animals (p < 0.001). Immunohistochemical analysis revealed enhanced expression of TGF- and -SMA in ethanol-cerulein mice compared to the saline group.
Conclusions: Intraperitoneal (IP) injections of ethanol and cerulein could successfully induce CP in mice. IP injections of ethanol provide higher reproducibility compared to ethanol feeding. The model is simple, non-invasive, reproducible, and time-saving. Since the protocol mimics the initial phases of CP development in alcoholics, it can be used for investigating basic mechanisms and testing new therapies.
Material and methods: Thirty-two male C57BL/6 mice were randomly selected, divided into four groups, and treated intraperitoneally with saline (10 ml/kg, control group), ethanol (3 g/kg; 30% v/v), cerulein (50 µg/kg), or ethanol + cerulein, for six weeks. Histopathological and immunohistochemical assays for chronic pancreatitis index along with real-time PCR assessments for mRNA levels of inflammatory cytokines and fibrogenic markers were conducted to verify the CP induction.
Results: The results indicated that CP index (CPI) was significantly increased in ethanol-cerulein mice compared to the saline, ethanol, and cerulein groups (p < 0.001). Interleukin 1 (IL-1), tumor necrosis factor (TNF-), transforming growth factor (TGF-), -smooth muscle actin (-SMA), and myeloperoxidase activity were also significantly greater in both cerulein and ethanol-cerulein groups than in the saline treated animals (p < 0.001). Immunohistochemical analysis revealed enhanced expression of TGF- and -SMA in ethanol-cerulein mice compared to the saline group.
Conclusions: Intraperitoneal (IP) injections of ethanol and cerulein could successfully induce CP in mice. IP injections of ethanol provide higher reproducibility compared to ethanol feeding. The model is simple, non-invasive, reproducible, and time-saving. Since the protocol mimics the initial phases of CP development in alcoholics, it can be used for investigating basic mechanisms and testing new therapies.
REFERENCES (47)
1.
Brock C, Nielsen LM, Lelic D, et al. (2013): Pathophysiology of chronic pancreatitis. World J Gastroenterol 19: 7231-7240.
2.
Ahmed SA, Wray C, Rilo HL, et al. (2006): Chronic pancreatitis: recent advances and ongoing challenges. Curr Probl Surg 43: 127-238.
3.
Ammann RW (2006): Diagnosis and management of chronic pancreatitis: current knowledge. Swiss Medical Weekly 136: 166-174.
4.
Behrman SW, Fowler ES (2007): Pathophysiology of chronic pancreatitis. Surg Clin North Am 87: 1309-1324.
5.
Dong F, Chen QQ, Zhuang ZH, et al. (2014): Multiple gene mutations in patients with type 2 autoimmune pancreatitis and its clinical features. Cent Eur J Immunol 39: 77-82.
6.
Siegmund SV, Haas S, Singer MV (2005): Animal models and their results in gastrointestinal alcohol research. Dig Dis 23: 181-194.
7.
Lerch MM, Gorelick FS (2013): Models of acute and chronic pancreatitis. Gastroenterology 144: 1180-1193.
8.
Saluja AK, Dudeja V (2013): Relevance of Animal Models of Pancreatic Cancer and Pancreatitis to Human Disease. Gastroenterology 144: 1194-1198.
9.
Otsuki M, Yamamoto M, Yamaguchi T (2010): Animal models of chronic pancreatitis. Gastroenterol Res Pract 2010: 403295.
10.
Olesen AE, Brokjaer A, Fisher IW, et al. (2013): Pharmacological challenges in chronic pancreatitis. World J Gastroenterol 19: 7302-7307.
11.
Clemens DL, Wells MA, Schneider KJ, et al. (2014): Molecular mechanisms of alcohol associated pancreatitis. World.
13.
Witt H, Apte MV, Keim V, et al. (2007): Chronic pancreatitis: challenges and advances in pathogenesis, genetics, diagnosis, and therapy. Gastroenterology 132: 1557-1573.
14.
Zhao JB, Liao DH, Nissen TD (2013): Animal models of pancreatitis: can it be translated to human pain study? World J Gastroenterol 19: 7222-7230.
15.
Nepomnyashchikh LM, Lushnikova EL, Viskunov VG, et al. (2011): Ultrastructure of acinar cell injuries in experimental acute pancreatitis created by common bile duct ligation. Bull Exp Biol Med 150: 747-752.
16.
Aghdassi AA, Mayerle J, Christochowitz S, et al. (2011): Animal models for investigating chronic pancreatitis. Fibrogenesis Tissue Repair 4: 26.
17.
Moloudi MR, Hasanzadeh K, Rouhani S, et al. (2014): Effect of chloroformic extract of Cichorium intybus on liver function tests and serum level of TNF- in obstructive cholestasis in rat. Scientific Journal of Kurdistan University of Medical Sciences 19: 10-19.
18.
Jerrells TR, Chapman N, Clemens DL (2003): Animal model of alcoholic pancreatitis: role of viral infections. Pancreas 27: 301-304.
19.
Elsasser HP, Haake T, Grimmig M, et al. (1992): Repetitive Cerulein-Induced Pancreatitis and Pancreatic Fibrosis in the Rat. Pancreas 7: 385-390.
20.
Kim H (2008): Cerulein Pancreatitis: Oxidative Stress, Inflammation, and Apoptosis. Gut and Liver 2: 74-80.
21.
Neuschwander-Tetri BA, Burton FR, Presti ME, et al. (2000): Repetitive self-limited acute pancreatitis induces pancreatic fibrogenesis in the mouse. Dig Dis Sci 45: 665-674.
22.
Kono H, Nakagami M, Rusyn I, et al. (2001): Development of an animal model of chronic alcohol-induced pancreatitis in the rat. Am J Physiol Gastrointest Liver Physiol 280: G1178-86.
23.
Fakhari S, Abdolmohammadi K, Panahi Y, et al. (2014): Glycyrrhizin attenuates tissue injury and reduces neutrophil accumulation in experimental acute pancreatitis. Int J Clin Exp Pathol 7: 101-109.
24.
Hassanzadeh A, Shahvaisi K, Hassanzadeh K, et al. (2015): Effects of rebamipide and encapsulating rebamipide with chitosan capsule on inflammatory mediators in rat experimental colitis. Sci J Kurdistan Univ Med Sci 20: 94-104.
25.
Bai H, Chen X, Zhang L, et al. (2012): The effect of sulindac, a non-steroidal anti-inflammatory drug, attenuates inflammation and fibrosis in a mouse model of chronic pancreatitis. BMC Gastroenterol 12: 115.
26.
Nikkhoo B, Jalili A, Fakhari S, et al. (2014): Nuclear pattern of CXCR4 expression is associated with a better overall survival in patients with gastric cancer. J Oncol 2014: 808012.
27.
Shafie A, Moradi F, Izadpanah E, et al. (2015): Neuroprotection of donepezil against morphine-induced apoptosis is mediated through Toll-like receptors. Eur J Pharmacol 764: 292-297.
28.
Shirzad H, Bagheri N, Azadegan-Dehkordi F, et al. (2015): New insight to IL-23/IL-17 axis in Iranian infected adult patients with gastritis: effects of genes polymorphisms on expression of cytokines. Acta Gastroenterol Belg 78: 212-218.
29.
Ahmadi A, Zandi F, Gharib A, et al. (2013): Relationship between polymorphism in promoter region of E-Cadherin (Cdh1) gene and helicobacter pylori infection in Kurdish population of Iran. Life Sci J 10: 552-556.
30.
Vonlaufen A, Wilson JS, Pirola RC, et al. (2007): Role of alcohol metabolism in chronic pancreatitis. Alcohol Res Health 30: 48-54.
31.
Sun Y, Xing GM, Bai J, et al. (2014): A New Mouse Model of Chronic Pancreatitis in C57BL/6J Strain That Mimics the Human Pathology. Pancreas 43: 148-150.
32.
Wan MH, Huang W, Latawiec D, et al. (2012): Review of experimental animal models of biliary acute pancreatitis and recent advances in basic research. HPB (Oxford) 14: 73-81.
33.
Singh S, Kumar R, Choudhuri G, et al. (2012): Chronic pancreatitis: A new pathophysiology. Indian J Hum Genet 18: 380-382.
34.
Lysy PA, Weir GC, Bonner-Weir S (2012): Concise review: pancreas regeneration: recent advances and perspectives. Stem Cells Transl Med 1: 150-159.
35.
Haber P, Nakamura M, Tsuchimoto K, et al. (2001): Alcohol and the pancreas. Alcohol Clin Exp Res 25: 244S-250S.
36.
Pandol SJ, Gukovsky I, Satoh A, et al. (2003): Animal and in vitro models of alcoholic pancreatitis: role of cholecystokinin. Pancreas 27: 297-300.
37.
Perides G, Tao X, West N, et al. (2005): A mouse model of ethanol dependent pancreatic fibrosis. Gut 54: 1461-1467.
38.
Pohlers D, Brenmoehl J, Loffler I, et al. (2009): TGF-beta and fibrosis in different organs - molecular pathway imprints. Biochim Biophys Acta 1792: 746-756.
39.
Gordon KJ, Blobe GC (2008): Role of transforming growth factor-beta superfamily signaling pathways in human disease. Biochim Biophys Acta 1782: 197-228.
40.
Omary MB, Lugea A, Lowe AW, et al. (2007): The pancreatic stellate cell: a star on the rise in pancreatic diseases. J Clin Invest 117: 50-59.
41.
Tahara H, Sato K, Yamazaki Y, et al. (2013): Transforming growth factor-alpha activates pancreatic stellate cells and may be involved in matrix metalloproteinase-1 upregulation. Lab Invest 93: 720-732.
42.
Miralles F, Battelino T, Czernichow P, et al. (1998): TGF-beta plays a key role in morphogenesis of the pancreatic islets of Langerhans by controlling the activity of the matrix metalloproteinase MMP-2. J Cell Biol 143: 827-836.
43.
Woods LT, Camden JM, El-Sayed FG, et al. (2015): Increased Expression of TGF-beta Signaling Components in a Mouse Model of Fibrosis Induced by Submandibular Gland Duct Ligation. PLoS One 10: e0123641.
44.
Lee H, Lim C, Lee J, et al. (2008): TGF-beta signaling preserves RECK expression in activated pancreatic stellate cells. J Cell Biochem 104: 1065-1074.
45.
Schneider L, Dieckmann R, Hackert T, et al. (2014): Acute alcohol-induced pancreatic injury is similar with intravenous and intragastric routes of alcohol administration. Pancreas 43: 69-74.
46.
Falzon M, Bhatia V (2015): Role of parathyroid hormone-related protein signaling in chronic pancreatitis. cancers (Basel) 7: 1091-1108.
47.
Schneider KJ, Scheer M, Suhr M, et al. (2012): Ethanol administration impairs pancreatic repair after injury. Pancreas 41: 1272-1279.
| eISSN: | 1644-4124 |
| ISSN: | 1426-3912 |
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