Biliary pancreatitis-associated ascitic fluid activates the production of tumor necrosis factor-alpha in acinar cells.
OBJECTIVE
Acute pancreatitis is associated with increased cytokine release from different cell sources. We have investigated the ability of acinar cells, in comparison with inflammatory peripheral blood cells, to produce tumor necrosis factor (TNF)-alpha in response to pancreatitis-associated ascitic fluid (PAAF).


METHODS
Controlled, randomized animal study.


METHODS
University research laboratory.


METHODS
Male Wistar rats.


METHODS
Flow cytometry using phycoerythrin-labeled monoclonal anti-TNF-alpha antiserum.


RESULTS
PAAF (20%, v:v) obtained from rats with acute pancreatitis induced by bile-pancreatic duct obstruction significantly increased TNF-alpha production in acinar cells, as measured by flow cytometry using phycoerythrin-labeled monoclonal anti-TNF-alpha antiserum. Neither heating of PAAF nor the addition of soybean trypsin inhibitor or neutralizing amounts of anti-TNF-alpha monoclonal antiserum reduced the acinar cell TNF-alpha production. Monocytes and lymphocytes did not produce TNF-alpha in response to PAAF. Likewise, the typical monocyte and lymphocyte stimulating factors-lipopolysaccharide (10 microg/microL) and phorbol 12-myristate 13-acetate (250 ng/mL) plus ionomycin (1 microg/mL), respectively-were not able to produce TNF-alpha in acinar cells. By comparison of the two acinar cell populations differentiated by flow cytometry, R2 cells (with higher forward scatter values) showed a greater ability to produce TNF-alpha in response to PAAF than R1 cells. Acinar cell nuclear factor-kappaB was activated, but TNF-alpha production was not totally inhibited in presence of N-acetyl cysteine (30, 100 mM).


CONCLUSIONS
The production of TNF-alpha from different cell sources is selectively activated. PAAF may be involved in the pathophysiology of acute pancreatitis by TNF-alpha production in acinar cells through mechanisms partially mediated by nuclear factor-kappaB activation. PAAF components, such as TNF-alpha or trypsin, are not responsible for acinar cell activation. TNF-alpha was induced by heat-resistant PAAF factors, displaying acinar cells with higher forward scatter (R2) a greater ability to increase the TNF-alpha production than R1 cells.
Ramudo
Laura
L
Department of Physiology and Pharmacology. University of Salamanca, Salamanca. Spain.
Manso
Manuel A
MA
De Dios
Isabel
I
D000208:Acute Disease; D000818:Animals; D001202:Ascitic Fluid / Q000276:immunology*; D001651:Cholestasis, Extrahepatic / Q000276:immunology*; D016207:Cytokines / Q000378:metabolism; D004195:Disease Models, Animal; D042783:Endothelial Cells / Q000276:immunology; D005434:Flow Cytometry; D008214:Lymphocytes / Q000276:immunology; D008297:Male; D009000:Monocytes / Q000276:immunology; D016328:NF-kappa B / Q000378:metabolism; D046790:Pancreas, Exocrine / Q000276:immunology*; D051381:Rats; D017208:Rats, Wistar; D018746:Systemic Inflammatory Response Syndrome / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*
D016207:Cytokines; D016328:NF-kappa B; D014409:Tumor Necrosis Factor-alpha