ABSTRACT
BACKGROUND: Severe hyperstimulation and duct obstruction pancreatitis (SHOP) is characterized by pancreatic fibrosis and loss of acinar cell mass. MMP-2 and MMP-9 are type IV collagenases and gelatinases. We hypothesized that fibrosis results from disruption of the normal collagen homeostasis and that altered activity of the type IV collagenases may contribute to pancreatic fibrosis in SHOP. METHODS: SHOP rats (n = 15) were prepared with pancreatic duct obstruction and cerulein (50 microg/kg/d, ip) hyperstimulation. Pancreas from unoperated control (n = 8), 48 h SHOP (n = 8), and 96 h SHOP (n = 7) rats was harvested, homogenized, and assayed for protein concentration (BCA method). Type IV collagenase (MMP-2 and MMP-9) expression was measured by zymography using gelatin as substrate. Type IV collagenase activity was quantified with a fluorescence assay. RESULTS: Expression of the active form of MMP-9 decreased while latent MMP-9 and active and latent MMP-2 increased on gelatin zymography. Activity of type IV collagenases (MMP-2 and MMP-9) progressively decreases with SHOP injury. The differences between expression and activity are likely due to posttranslational regulators such as MT-MMPs and TIMPs. CONCLUSIONS: Collagenase expression and activity are decreased in the SHOP model of pancreatitis, suggesting a decrease in the homeostatic mechanisms for type IV collagen in the extracellular matrix. Therefore, early fibrosis in the SHOP model is, at least in part, due to alterations in collagen homeostasis and not simply increased collagen production.
Subject(s)
Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/metabolism , Pancreas/enzymology , Pancreatitis/metabolism , Animals , Chronic Disease , Disease Models, Animal , Enzyme Activation , Extracellular Matrix/enzymology , Fibrosis , Gelatin , Homeostasis/physiology , Male , Pancreas/pathology , Pancreatitis/pathology , Rats , Rats, Sprague-DawleyABSTRACT
Significant fibrosis and acinar atrophy are characteristics of chronic pancreatitis; however, because of the lack of a reproducible model, early phases of these changes are poorly understood. We have developed a model of severe hyperstimulation and obstruction pancreatitis (SHOP) to better define the mechanisms of early pancreatic fibrogenesis. Sprague-Dawley rats were used and SHOP was induced by complete pancreatic duct obstruction and daily cerulein hyperstimulation (50 microg/kg intraperitoneally). Animals were killed at 24, 48, 72, and 96 hours. Control animals underwent sham operation and received no cerulein. Pancreata were prepared for hematoxylin and eosin and sirius red (collagen-specific) staining and for hydroxyproline assay (measure of total collagen content). We found moderate amounts of edema and inflammation but minimal parenchymal necrosis. Significant loss of acinar cell mass was noted by 48 hours, and normal acinar cells were essentially absent by 96 hours. Tissue collagen content increased with time and large amounts of interstitial collagen were detected by 72 hours. In conclusion, SHOP is a novel model of early pancreatic fibrosis associated with minimal necrosis and a significant decrease in acinar cell mass, making it an ideal model to study the early cellular mechanisms of pancreatic fibrogenesis.
