Ameliorative effects of the melatonin on some cytokine levels, NF-κB immunoreactivity, and apoptosis in rats with cerulein-induced acute pancreatitis

Document Type : Original Article


1 University of Selçuk, Faculty of Veterinary Medicine, Department of Physiology, Turkey

2 University of Selçuk, Faculty of Veterinary Medicine, Department of Histology and Embryology, Turkey


Objective(s): Investigating the ameliorative effects of melatonin on cytokine levels, apoptosis, and NF-κB immunoreactivity in rats with cerulein-induced acute pancreatitis.
Materials and Methods: Thirthy-two Wistar Albino rats were divided into four groups: Control group which didn’t undergo acute pancreatitis induction and was left without treatment, pancreatitis group in which the acute pancreatitis was induced by 2 successive intraperitoneal doses of cerulein at a 2-hour interval (50 µg/kg and then 25 µg/kg), melatonin-treated pancreatitis group which was intraperitoneally administrated with 50 mg/kg of melatonin, 30 min before each cerulein injection, and melatonin group which was intraperitoneally administrated with 2 successive doses of melatonin (50 mg/kg each) at a 2-hour interval. Pancreatic tissue and blood samples were taken from animals of all groups. IL-1β, TNF-α, and IL-10 levels were determined in blood samples. Apoptosis was determined by the TUNEL assay and the NF-κB was detected immunohistochemically in acinar cells of the exocrine pancreatic portion.
Results: IL-1β, TNF-α, and IL-10 levels in the acute pancreatitis group were significantly increased when compared to the control negative group. IL-1β and TNF-α levels in the melatonin-treated pancreatitis group were significantly lower than those of the acute pancreatitis group. While number of apoptotic cells and percentage of NF-κB immunopositive cells in the acute pancreatitis group were significantly increased compared to other groups and it was observed that these parameters were significantly reduced in the melatonin-treated pancreatitis group compared to the acute pancreatitis group.
Conclusion: These findings suggest that melatonin administration can significantly reduce the severity of acute pancreatitis in rats.


Main Subjects

1. Stevenson K, Carter CR. Acute pancreatitis. Surgery 2013; 31: 295-303. 
2. Richard W, Nelson C, Couto G. Small Animal Internal Medicine (5th edition). by mosby, St. Louis Missouri; 2014.
3. Özkan E, Akyüz C, Dulundu E, Topaloğlu U, Sehirli AÖ, Ercan F, et al. Protective effects of lycopene on cerulein-induced experimental acute pancreatitis in rats. J Surg Res 2012; 176: 232-238. 
4. Petrov MS. Therapeutic implications of oxidative stress in acute and chronic pancreatitis. Curr Opin Clin Nutr Metab Care 2010; 13: 562-568. 
5. Carrasco C, Marchena AM, Holguín-Arévalo MS, Martín-Partido G, Rodríguez AB, Paredes SD, et al. Anti-inflammatory effects of melatonin in a rat model of caerulein-induced acute pancreatitis. Cell Biochem Func 2013; 31: 585-590. 
6. Jaworek J, Bonio J, Leja-Szpa A, Nawrot K, Tomaszewska MR, Stachura J, et al. Sensory nerves in central and peripheral control of pancreatic integrity by leptin and melatonin. J Physiol Pharmacol 2002; 53: 51-74. 
7. Jung KH, Hong SW, Zheng HM, Lee HS, Lee H, Lee DH, et al. Melatonin ameliorates cerulein-induced pancreatitis by modulation of nuclear erythroid 2-related factor 2 and nuclear factor-kappaB in rats. J Pineal Res 2010; 48: 239-250. 
8. Qi W, Tan DX, Reiter RJ, Kim SJ, Manchester LC, Cabrera J, et al. Melatonin reduces lipid peroxidation and tissue edema in cerulein-induced acute pancreatitis in rats. Dig Dis Sci 1999; 44: 2257-2262. 
9. Yu JH, Lim JW, Kim KH, Morio T, Kim H. NADPH oxidase and apoptosis in cerulein-stimulated pancreatic acinar AR42J cells. Free Radic Biol Med 2005; 39: 590-602. 
10. Kim H. Cerulein pancreatitis: Oxidative stress, inflammation, and apoptosis. Gut and Liver 2008; 2: 74-80. 
11. Frossard JL, Hadengue A, Pastor CM. New serum markers for the detection of severe acute pancreatitis in humans. Am J Resp Crit Care Med 2001; 164: 162-170. 
12. Heath DI, Cruickshank A, Gudgeon M, Jehanli A, Shenkin A, Imrie CW. Role of interleukin-6 in mediating the acute phase protein response and potential as an early means of severity assessment in acute pancreatitis. Gut 1993; 34: 41-45. 
13. Sameshima H, Ikei S, Mori K, Yamaguchi Y, Egami H, Misumi M, et al. The role of tumor necrosis factor-alpha in the aggravation of cerulein-induced pancreatitis in rats. Int J Pancreatol 1993; 14: 107-115. 
14. Gukovskaya AS, Mareninova OA, Odinokova IV, Sung KF, Lugea A, Fischer L, et al. Cell death in pancreatitis: Effects of alcohol. J Gastroenterol Hepatol 2006; 21: S10-13. 
15. Liu Y, Zhou ZG, Zhou B, Wang R, Yan H, Li Y. Downregulation of GRP78 and XIAP is correlated with apoptosis during cerulein-induced acute pancreatitis in rats via regulation of caspase activation. Mol Med Rep 2013; 7: 725-730. 
16. Mareninova OA, Sung KF, Hong P, Lugea A, Pandol SJ, Gukovsky I, et al. Cell death in pancreatitis: Caspases protect from necrotizing pancreatitis. J Biol Chem 2006; 281: 3370-3381. 
17. Yu JH, Lim JW, Kim H, Kim KH. NADPH oxidase mediates interleukin-6 expression in cerulein-stimulated pancreatic acinar cells. Int J Biochem Cell Biol 2005b; 37: 1458-1469. 
18. Kim H. Inhibitory mechanism of lycopene on cytokine expression in experimental pancreatitis. Ann N Y Acad Sci 2011; 1229: 99-102. 
19. Babu BI, Malleo G, Genovese T, Mazzon E, Di Paola R, Crisafulli C, et al. Green tea polyphenols ameliorate pancreatic injury in cerulein-induced murine acute pancreatitis. Pancreas 2009; 38: 954-967. 
20. Yu JH, Kim H. Oxidative stress and inflammatory signaling in cerulein pancreatitis. World J Gastroenterol 2014; 20:17324-17329. 
21. Lerner AB, Case JD, Takahashi Y, Lee TH, Mori W. Isolation of melatonin, the pineal gland factor that lightens melanocytes. J Am Chem Soc 1958; 80: 2587. 
22. Jaworek J, Brzozowski T, Konturek SJ. Melatonin as an organoprotector in the stomach and the pancreas. J Pineal Res 2005; 38: 73-83. 
23. Chen CQ, Fichna J, Bashashati M, Li YY, Storr M. Distribution, function and physiological role of melatonin in the lower gut. World J Gastroenterol 2011; 17: 3888-3898. 
24. Jaworek J, Szklarczyk J, Jaworek AK, Nawrot-Porąbka K, Leja-Szpak A, Bonior J, et al. Protective effect of melatonin on acute pancreatitis. Int J Inflam 2012; 2012: 1-8. 
25. Hardeland R, Tan DX, Reiter RJ. Kynuramines, metabolites of melatonin and other indoles: the resurrection of an almost forgotten class of biogenic amines. J Pineal Res 2009; 47: 109-126. 
26. Jaworek J, Leja-Szpak A, Bonior J, Nawrot K, Tomaszewska R, Stachura J, et al. Protective effect of melatonin and its precursor L-tryptophan on acute pancreatitis induced by caerulein overstimulation or ischemia/reperfusion. J Pineal Res 2003; 34: 40-52. 
27. Sato S, Stark HA, Martinez J, Beaven MA, Jensen RT, Gardner JD. Receptor occupation, calcium mobilization, and amylase release in pancreatic acini: Effect of CCK-JMV-180. Am J Physiol 1989;257:202-209. 
28. Lerch MM, Adler G. Experimental animal models of acute pancreatitis. Int J Pancreatol 1994; 15:159-70. 
29. Özaydın T, Öznurlu Y, Sur E, Çelik İ, Uluışık D. The effects of bisphenol A on some plasma cytokine levels and distribution of CD8+ and CD4+ T lymphocytes in spleen, ileal Peyer’s patch and bronchus associated lymphoid tissue in rats. Acta Histochem 2018; 120: 728-733. 
30. Keskin E, Uluışık D, Öznurlu Y, Özaydın T. Effects of coenzyme Q10 on some blood antioxidant system parameters and histological changes in the pancreas and aorta of streptozotocin-induced diabetic rats. Kocatepe Vet J 2020; 13: 192-202.
31. Crossmon G. A modification of Mallory’s connective tissue stain with a discussion of the principles involved. Anat Rec 1937;69:33-38
32. Barouch LA, Gao D, Chen L, Miller KL, Xu W, Phan AC, et al. Cardiac myocyte apoptosis is associated with increased DNA damage and decreased survival in murine models of obesity. Circ Res 2006; 98:119-124. 
33. Özaydın T, Çelik İ. Histological, histochemical and immunohistochemical investigations on the developing small intestines of broiler embryos. J Anim Vet Adv 2012; 11: 2936-2944.
34. Sternby H, Hartman H, Thorlacius H, Regnér S. The initial course of IL1β, IL-6, IL-8, IL-10, IL-12, IFN-γ and TNF-α with regard to severity grade in acute pancreatitis. Biomolecules 2021; 11: 591-599. 
35. Ren K, Lin J, Wang Y, Ji X. Myricetin alleviates pancreatic microcirculation and inflammation in rats with severe acute pancreatitis through FOXO1 and NF-κB pathways. Mol Cell Toxicol 2023
36. Ali EM, Madkour HI. Effect of melatonin on experimentally induced acute pancreatitis and associated hyperlipidemia. Rec Pharm Biomedical Sci 2023; 7: 91-102. 
37. Zhao Q, Zhang H, Wu J, Lv X, Jin X, Hu J. Melatonin inhibits the endoplasmic reticulum stressinduced, C/EBP homologous proteinmediated pathway in acute pancreatitis. Mol Med Rep 2020; 22: 1647-1655. 
38. Le PH, Kuo CJ, Cheng HT, Wu RC, Chen TH, Lin CJ, et al. Melatonin attenuates acute pancreatitis-induced liver damage through Akt-dependent PPAR-γ pathway. J Surg Res 2019; 236: 311-318. 
39. Chen Y, Zhao Q, Chen Q, Zhang Y, Shao B, Jin Y, et al. Melatonin attenuated inflammatory reaction by inhibiting the activation of p38 and NFκB in taurocholateinduced acute pancreatitis. Mol Med Rep 2018; 17: 5934-5939. 
40. Grupp K, Erbes J, Poppe A, Wodack K, Gocht A, Trepte C, et al. Melatonin treatment of pigs with acute pancreatitis reduces inflammatory reaction of pancreatic tissue and enhances fitness score of pigs: experimental research. World J Emerg Surg 2019; 14: 18-25. 
41. Colares JR, Schemitt EG, Hartmann RM, Licks F, Soares MD, Bosco AD, et al. Antioxidant and anti-inflammatory action of melatonin in an experimental model of secondary biliary cirrhosis induced by bile duct ligation. World J Gastroenterol 2016; 22: 8918-8928. 
42. Jaworek J, Konturek SJ. Hormonal protection in acute pancreatitis by ghrelin, leptin and melatonin. World J Gastroenterol 2014; 20: 16902-16912. 
43. Jaworek J, Konturek SJ, Tomaszewska R, Leja-Szpak A, Bonior J, Nawrot K, et al. The circadian rhythm of melatonin modulates the severity of caerulein-induced pancreatitis in the rat. J Pineal Res 2004; 37: 161-170. 
44. Eşrefoğlu M, Gül M, Ateş B, Selimoğlu MA. Ultrastructural clues for the protective effect of melatonin against oxidative damage in cerulein-induced pancreatitis. J Pineal Res 2006; 40: 92-97. 
45. Sadek AS, Khattab RT. The protective role of melatonin on L-arginine-induced acute pancreatitis in adult male albino rats. Folia Morphol (Warsz) 2017; 76: 66-73. 
46. Motilva V, García-Mauriño S, Talero E, Illanes M. New paradigms in chronic intestinal inflammation and colon cancer: role of melatonin. J Pineal Res 2011; 51: 44-60. 
47. Laliena A, San Miguel B, Crespo I, Alvarez M, González-Gallego J, Tuñón MJ. Melatonin attenuates inflammation and promotes regeneration in rabbits with fulminant hepatitis of viral origin. J Pineal Res 2012; 53: 270-278. 
48. Eşrefoğlu M, Gül M, Ates B, Batçioğlu K, Selimoğlu MA. Antioxidative effect of melatonin, ascorbic acid and N-acetylcysteine on caerulein-induced pancreatitis and associated liver injury in rats. World J Gastroenterol 2006; 12: 259-264. 
49. Chen H, Wang L, Xing BZ, Liu XH, Chen ZY, Weng XD, et al. Ischemic postconditioning attenuates inflammation in rats following renal ischemia and reperfusion injury. Exp Ther Med 2015; 10: 513-518. 
50. Gukovsky I, Gukovskaya AS, Blinman TA, Zaninovic V, Pandol SJ. Early NF-kappaB activation is associated with hormone-induced pancreatitis. Am J Physiol 1998; 275: G1402-1414. 
51. Huang H, Liu Y, Daniluk J, Gaiser S, Chu J, Wang H, et al. Activation of nuclear factor-κB in acinar cells increases the severity of pancreatitis in mice. Gastroenterology 2013; 144: 202-210. 
52. Bansod S, Godugu C. Nimbolide ameliorates pancreatic inflammation and apoptosis by modulating NF-κB/SIRT1 and apoptosis signaling in acute pancreatitis model. Int Immunopharmacol 2021; 90: 107246. 
53. Zhang D, Li L, Li J, Wei Y, Tang J, Man X, et al. Colchicine improves severe acute pancreatitis-induced acute lung injury by suppressing inflammation, apoptosis and oxidative stress in rats. Biomed Pharmacother 2022; 153: 113461-113470. 
54. Chen H, Sun YP, Li Y, Liu WW, Xiang HG, Fan LY, et al. Hydrogen-rich saline ameliorates the severity of l-arginine-induced acute pancreatitis in rats. Biochem Biophys Res Comm 2010; 393: 308-313. 
55. Szabolcs A, Reiter RJ, Letoha T, Hegyi P, Papai G, Varga I, et al. Effect of melatonin on the severity of L-arginine-induced experimental acute pancreatitis in rats. World J Gastroenterol 2006; 12: 251-258. 
56. Huai J, Shao Y, Sun X, Jin Y, Wu J, Huang Z. Melatonin ameliorates acute necrotizing pancreatitis by the regulation of cytosolic Ca2+ homeostasis. Pancreatology 2012; 12: 257-263. 
57. Gülben K, Ozdemir H, Berberoğlu U, Mersin H, Yrkin F, Cakýr E, et al. Melatonin modulates the severity of taurocholate-induced acute pancreatitis in the rat. Dig Dis Sci 2010; 55: 941-946. 
58. Jaworek J, Leja-Szpak A, Nawrot-Porąbka K, Szklarczyk J, Kot M, Pierzchalski P, et al. Effects of melatonin and its analogues on pancreatic inflammation, enzyme secretion, and tumorigenesis. Int J Mol Sci 2017; 18: 1014-1026. 
59. Muñoz-Casares FC, Padillo FJ, Briceño J, Collado JA, Muñoz-Castañeda JR, Ortega R, et al. Melatonin reduces apoptosis and necrosis induced by ischemia/reperfusion injury of the pancreas. J Pineal Res 2006; 40: 195-203.