Abstract
The Fas/Fas ligand (FasL) system is suggested to be correlated to the onset of inflammation and apoptosis in various diseases. However, whether Fas and FasL are expressed in chronic pancreatitis is unknown. The aim of this study was to examine the expression of the Fas/FasL system and to analyze its correlation with apoptosis in a spontaneous chronic pancreatitis model (the WBN/Kob rat). Four-week-old male WBN/Kob rats were fed a special pellet diet (MB-3). Different groups of rats were killed every four weeks, and pancreata were histopathologically examined. Fas and FasL mRNAs in the pancreas were detected with a reverse transcription-polymerase chain reaction method. The cellular localization of Fas and FasL mRNA and protein was determined with in situ hybridization (ISH) and immunohistochemistry (IHC). Apoptosis was detected with a terminal deoxynucleotidyltransferase-mediated method. Fas and FasL mRNA were expressed when the pancreas was still pathologically normal, and showed a biphasic peak at 12 and 20 weeks. ISH and IHC confirmed that Fas and FasL are expressed in the cytoplasm of acinar cells, ductal cells, and lymphocytes. An apoptotic index in acinar cells correlated to the expression of Fas and FasL mRNAs. These results suggest that the expression of the Fas/FasL system is involved in acinar cell apoptosis and the onset and progression of chronic pancreatitis in the WBN/Kob rat.
Similar content being viewed by others
REFERENCES
Ito N, Yonehara S, Ishii A, Yonehara M, Mizushima S, Sameshima M, Atsushi H, Seto Y, Nagata S: The polypeptide encoded by the cDNA for human surface antigen Fas can mediate apoptosis. Cell 66:233–243, 1991
Oehm A, Behrmann I, Falk W, Pawlita M, Maier G, Klas C, Li-Weber M, Richards S, Dhein J, Trauth BC, Ponstingl H, Krammer PH: Purification and molecular cloning of the APO-1 cell surface antigen, a member of the tumor necrosis factor/nerve growth factor receptor superfamily. J Biol Chem 267:10709–10715, 1992
Suda T, Takahashi T, Golstein P, Nagata S: Molecular cloning and expression of the Fas ligand, a novel member of the tumor necrosis factor family. Cell 75:1169–1178, 1993
Suda T, Nagata S: Purification and characterization of the Fas-ligand that induces apoptosis. J Exp Med 179:873–879, 1994
Galle PR, Hofmann WJ, Walczak H, Schaller H, Otto G, Stremmel W, Krammer PH, Runkel L: CD95 (Apo-1/Fas) receptor and ligand in liver damage. J Exp Med 182:1223–1230, 1995
Hayashi N, Mita E: Fas system and apoptosis in viral hepatitis. J Gastroenterol Hepatol 12:S223–S226, 1997
Hiramatsu N, Hayashi N, Katayama K, Mochizuki K, Kawanishi Y, Kasahara A, Fusamoto H, Kamada T: Immunohistochemical detection of Fas antigen in liver tissue of patients with chronic hepatitis C. Hepatology 19:1354–1359, 1994
Cheng J, Zhou T, Liu C, Shapiro JP, Brauer MJ, Kiefer MC, Barr PJ, Mountz JD: Protection from Fas-mediated apoptosis by a soluble form of the Fas molecule. Science 263:1759–1762, 1994
Siegel RM, Fleisher TA: The role of Fas and related death receptors in autoimmune and other disease states. J Allergy Clin Immunol 103:729–738, 1999
Chervonsky AV, Wang Y, Wong FS, Visintin I, Flavell RA, Janeway Jr CA, Matis LA: The role of Fas in autoimmune diabetes. Cell 89:17–24, 1997
Yasuda H, Kataoka K, Ichimura H, Mitsuyoshi M, Iida T, Kida M, Imanishi J: Cytokine expression and induction of acinar cell apoptosis after pancreatic duct ligation in mice. J Interferon Cytokine Res 19:637–644, 1999
Bernstorff WV, Spanjaard RA, Chan AK, Lockhart DC, Sadanaga N, Wood I, Peiper M, Goedegebuure PS, Eberlein TJ: Pancreatic cancer cells can evade immune surveillance via nonfunctional Fas (Apo-1/CD95) receptors and aberrant expression of functional Fas ligand. Surgery 125:73–84, 1999
Su S-B, Motoo Y, Xie M-J, Sakai H, Taga H, Sawabu N: Expression of pancreatitis-associated protein (PAP) in spontaneous chronic pancreatitis in the WBN/Kob rat: Effect of herbal medicine Saiko-keishi-to (TJ-10). Pancreas 19:239–247, 1999
Su S-B, Motoo Y, Xie M-J, Miyazono K, Sawabu N: Expression of transforming growth factor beta in spontaneous chronic pancreatitis in the WBN/Kob rat. Dig Dis Sci45:151–159, 2000
Kakinuma C, Suda K, Shibutani Y: Histopathological and biochemical studies on pancreatic fibrosis in WBN/Kob rats. Virchows Arch 434:83–89, 1999
Searle J, Kerr JFR, Bishop CJ: Necrosis and apotosis: distant modes of cell death with fundamentally different significance. Pathol Annu 17:229–259, 1982
Walker NI: Ultrastructure of the rat pancreas after experimental duct ligation. I. The role of apoptosis and intraepithelial macrophages in acinar cell deletion. Am J Pathol 126:439–451, 1987
Walker NI, Winterford CM, Kerr JFR: Ultrastructure of the rat pancreas after experimental duct ligation. II. Duct and stromal cell proliferation, differentiation, and deletion. Pancreas 7:420–434, 1992
Gukovskaya AS, Perkins P, Zaninovic V, Sandoval D, Rutherford R, Fitzsimmons T, Pandol SJ, Poucell-Hatton S: Mechanisms of cell death after pancreatic duct obstruction in the opossum and rat. Gastroenterology 110:875–884, 1996
Rao MS, Yeldandi AV, Subbarao, Reddy JK: Role of apoptosis in copper deficiency-induced pancreatic involution in the rat. Am J Pathol 142:1952–1957, 1993
Ide H, Yeldandi AV, Reddy JL, Rao MS: Increased expression of sulfated glycoprotein-2 and DNA fragmentation in the pancreas of copper-deficient rat. Toxicol Appl Pharmacol 126:174–177, 1994
Kazacos EA, Van Vleet JF: Sequential ultrastructural changes of the pancreas in zinc toxicosis in ducklings. Am J Pathol 134:581–595, 1989
Walkar NI, Winterford CM, Williamson RM, Kerr JFR: Ethionin-induced atrophy of rat pancreas involves apoptosis of acinar cells. Pancreas 8:443–449, 1993
Horne WI, Tsukamoto H: Dietary modulation of alcoholinduced pancreatic injury. Alcohol 10:481–484, 1993
Kaiser AM, Saluja AK, Sengupta A, Saluja M, Steer ML: Relationship between severity, necrosis, and apoptosis in five models of experimental acute pancreatitis. Am J Physiol 269:C1295–C1304, 1995
Kaiser AM, Saluja AK, Lu L, Yamanaka K, Yamaguchi Y, Steer ML: Effects of cycloheximide on pancreatic endonuclease activity, apoptosis, and severerity of acute pancreatitis. Am J Physiol 271:C982–C993, 1996
Saluja A, Hofbauer B, Yamaguchi Y, Yamanaka K, Steer ML: Induction of apoptosis reduces the severity of caeruleininduced pancreatitis in mice. Biochem Biophys Res Commun 220:875–878, 1996
Sandoval D, Gukovskaya A, Reavey P, Gukovsky S, Sisk A, Braquet P, Pandol SJ, Poucell-Hatton S: The role of neutrophils and platelet-activating factor in mediating experimental pancreatitis. Gastroenterology 111:1081–1091, 1995
Arai I, Hasegawa M, Mase A, Kitani T, Komatsu Y, Yamaura H, Sasaki K, Taguchi S: Development of pancreatic function and pathohistology in WBN/Kob rats using a special breeding diet. Jpn J Appl Physiol 27:373–380, 1997
Gavrieli Y, Sherman Y, Ben-Sasson SA: Identification of programmed cell death in situ via specific-labeling of nuclear DNA fragmentation. J Cell Biol 119:493–501, 1992
Yano H, Fukuda K, Haramaki M, Momosaki S, Ogasawara S, Higaki K, Kojiro M: Expression of Fas and anti-Fas mediated apoptosis in human hepatocellular cancinoma cell lines. J Hepatol 25:454–464, 1996
Krammer PH, Behrmann I, Daniel P, Dhein J, Debatin KM: Regulation of apoptosis in the immune system. Curr Opin Immunol 6:279–289, 1994
Brunner T, Mogil RJ, LaFace D, Yoo NJ, Mahboubi A, Echeverri F, Martin SJ, Force WR, Lynch DH, Ware CF, Green DR: Cell-autonomous Fas (CD95)/Fas-ligand interaction mediates activation-induced apoptosis in T-cell hybridomas. Nature 373:441–444, 1995
Lowin B, Hahne M, Mattmann C, Tschopp J: Cytolytic T cell cytotoxicity is mediated through perforin and Fas lytic pathways. Nature 370:650–652, 1995
Spielman J, Lee RK, Podack ER: Perforin/FasL double deficiency is associated with macrophage expansion and pancreatitis. J Immunol 161:7063–7070, 1998
Seino K, Tortolina G, Amelotti M, Dallegri F: Soluble Fas ligand is chemotactic for human neutrophilic polymorphonuclear leukocytes. J Immunol 161:4484–4488, 1998
Ottonello L, Iwabuchi K, Kayagaki N, Miyata R, Natsuzawa A, Fukao K, Yagita H, Okumura K: Chemotactic activity of soluble Fas ligand against phagocytes. J Immunol 162:3601–3606, 1999
Miwa K, Asano M, Horai R, Iwakura Y, Nagata S, Suda T: Caspase 1-independent IL-1beta release and inflammation induced by the apoptosis inducer Fas ligand. Nat Med 4:1287–1292, 1998
Chen JJ, Sun Y, Nabel GJ: Regulation of the proinflammatory effects of Fas ligand (CD95L). Science 282:1714–1717, 1998
Gukovskaya AS, Gukovsky I, Zaninovic V, Song M, Sandoval D, Gukovsky S, Pandol SJ: Pancreatic acinar cells produce, release, and respond to tumor necrosis factor-a. Role in regulating cell death and pancreatitis. J Clin Invest 100:1853–1862, 1997
Leithäuser F, Dhein J, Mechtersheimer G, Koretz K, Brüderlein S, Henne C, Schmidt A, Debatin K-M, Krammer PH, Möller P: Constitutive and induced expression of APO-1, a new member of the nerve growth factor/tumor necrosis factor receptor superfamily, in normal and neoplastic cells. Lab Invest 69:415–429, 1993
Müschen M, Warskulat U, Peters-Regehr T, Bode JG, Kubitz R, Häussinger D: Involvement of CD95 (Apo-1/Fas) ligand expressed by rat Kupffer cells in hapatic immunoregulation. Gastroenterology 116:666–677, 1999
Kayagaki N, Kawasaki A, Ebata T, Ohmoto H, Ikeda S, Inoue S, Yoshino K, Okumura K, Yagita H: Metalloproteinasesmediated release of human Fas ligand. J Exp Med 182:1777–1783, 1995
Tanaka M, Suda T, Haze K, Nakamura N, Sato K, Kimura F, Mizuki M, Tagawa S, Ohga S, Hataka K, Drummond AH, Nagata S: Fas ligand in human serum. Nat Med 2:317–322, 1996
Ishihara T, Hayasaka A, Yamaguchi T, Kondo F, Saisho H: Immunohistochemical study of transforming growth factor b1, matrix metalloproteinases-2,9, tissue inhibitors of metalloproteinases-1,2, and basement membrane components at pancreatic ducts in chronic pancreatitis. Pancreas 17:412–418, 1998
Müller-Pillasch F, Gress TM, Yamaguchi H, Geng M, Adler G, Menke A: The influence of transforming growth factor b1 on the expression of genes coding for matrix metalloproteinases and tissue inhibitors of metalloproteinases during regeneration from cerulein-induced pancreatitis. Pancreas 15:168–175, 1997
Kerr JFR, Wyllie AH, Currie AR: Apoptosis: a basic biological phenomenon with wideranging implications in tissue kinetics. Br J Cancer 26:239–257, 1972
Wyllie AH, Kerr JFR, Currie AR: Cell death: the significance of apoptosis. Int Rev Cytol 68:251–306, 1980
Lechêne de la Porte P, Iovanna JL, Odaira C, Odaira C, Choux R, Sarles H, Berger Z: Involvement of tubular complexes in pancreatic regeneration after necrohemorrhagic pancreatitis. Pancreas 6:298–306, 1991
Ludwig CU, Menke A, Adler G, Lutz MP: Fibroblasts stimulate acinar cell proliferation through IGF-I during regeneration from pancreatitis. Am J Physiol 276:G193–G198, 1999
Menke A, Yamaguchi H, Giehl K, Adler G. Hepatocyte growth factor and fibroblast growth factor 2 are overexpressed after cerulein-induced acute pancreatitis. Pancreas 18:28–33, 1999
Ebert M, Yokoyama M, Ishiwata T, Friess H, Büchler MW, Malfertheiner P, Korc M: Alteration of fibroblast growth factor and receptor expression after acute pancreatitis in humans. Pancreas 18:240–246, 1999
Sakagami J, Kataoka K, Ohta A, Nakajima T: Relationship of plasma CCK to acinar cell regeneration in acute pancreatitis as studied by proliferating cell nuclear antigen. Dig Dis Sci 41:1828–1837, 1996
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Su, SB., Motoo, Y., Xie, MJ. et al. Apoptosis in Rat Spontaneous Chronic Pancreatitis. Dig Dis Sci 46, 166–175 (2001). https://doi.org/10.1023/A:1005537218459
Issue Date:
DOI: https://doi.org/10.1023/A:1005537218459