deoxycholic-acid and Pancreatitis

Topics: Acute Disease; Animals; Bites and Stings; Cell Membrane; Complement C3; Complement System Proteins; Deoxycholic Acid; Edema; Humans; Immune Complex Diseases; Pancreas; Pancreatitis; Scorpions; Snake Venoms; Taurocholic Acid

Recent studies have demonstrated that dysfunction of the intestinal barrier is a significant contributing factor to the development of severe acute pancreatitis (SAP). A stable intestinal mucosa barrier functions as a major anatomic and functional barrier, owing to the balance between intestinal epithelial cell (IEC) proliferation and apoptosis. There is some evidence that calcium overload may trigger IEC apoptosis and that calcineurin (CaN)/nuclear factor of activated T-cells (NFAT) signaling might play an important role in calcium-mediated apoptosis.. To investigate the potential mechanisms underlying the therapeutic effect of Qingyi decoction (QYD) in SAP.. A rat model of SAP was created. Retrograde infusion of sodium deoxycholate increased the severity of pancreatic and intestinal pathology and the levels of serum amylase, TNF-α, and IL-6. Both the indicators of intestinal mucosa damage (D-lactic acid and DAO) and the levels of IEC apoptosis were elevated in the SAP group. QYD treatment reduced the serum levels of amylase, TNF-α, IL-6, D-lactic acid, and DAO and attenuated the histological findings. IEC apoptosis associated with SAP was ameliorated under QYD treatment. In addition, the protein expression levels of the two subunits of CaN were remarkably elevated in the SAP group, and the NFATc3 gene was significantly upregulated at both the transcript and protein levels in the SAP group compared with the control group. QYD significantly restrained CaN and NFATc3 gene expression in the intestine, which was upregulated in the SAP group. Furthermore, QYD serum significantly decreased the LPS-induced elevation in intracellular free Ca

Topics: Acute Disease; Amine Oxidase (Copper-Containing); Amylases; Animals; Caco-2 Cells; Calcineurin; Calcium; Deoxycholic Acid; Drugs, Chinese Herbal; Epithelial Cells; Humans; Interleukin-6; Intestinal Mucosa; Lactic Acid; Lipopolysaccharides; Pancreatitis; Rats; Rats, Sprague-Dawley; T-Lymphocytes; Tumor Necrosis Factor-alpha

To investigate the effect of Qingyi decoction on the expression of secreted phospholipase A2 (sPLA2) in intestinal barrier injury.. Fifty healthy Sprague-Dawley rats were randomly divided into control, severe acute pancreatitis (SAP), Qingyi decoction-treated (QYT), dexamethasone-treated (DEX), and verapamil-treated (VER) groups. The SAP model was induced by retrograde infusion of 1.5% sodium deoxycholate into the biliopancreatic duct of the rats. All rats were sacrificed 24 h post-SAP induction. Arterial blood, intestine, and pancreas from each rat were harvested for investigations. The levels of serum amylase (AMY) and diamine oxidase (DAO) were determined using biochemical methods, and serum tumor necrosis factor (TNF)-α level was measured by an enzyme linked immunosorbent assay. Pathologic changes in the harvested tissues were investigated by microscopic examination of hematoxylin and eosin-stained tissue sections. The expressions of sPLA2 at mRNA and protein levels were detected by reverse transcriptase PCR and Western blot, respectively. A terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay was used to investigate apoptosis of epithelial cells in the intestinal tissues.. Compared to the control group, the expression of sPLA2 at both the mRNA and protein levels increased significantly in the SAP group (0.36 ± 0.13 vs 0.90 ± 0.38, and 0.16 ± 0.05 vs 0.64 ± 0.05, respectively; Ps < 0.01). The levels of AMY, TNF-α and DAO in serum were also significantly increased (917 ± 62 U/L vs 6870 ± 810 U/L, 59.7 ± 14.3 ng/L vs 180.5 ± 20.1 ng/L, and 10.37 ± 2.44 U/L vs 37.89 ± 5.86 U/L, respectively; Ps < 0.01). The apoptosis index of intestinal epithelial cells also differed significantly between the SAP and control rats (0.05 ± 0.02 vs 0.26 ± 0.06; P < 0.01). The serum levels of DAO and TNF-α, and the intestinal apoptosis index significantly correlated with sPLA2 expression in the intestine (r = 0.895, 0.893 and 0.926, respectively; Ps < 0.05). The levels of sPLA2, AMY, TNF-α, and DAO in the QYT, VER, and DEX groups were all decreased compared with the SAP group, but not the control group. Qingyi decoction intervention, however, gave the most therapeutic effect against intestinal barrier damage, although the onset of its therapeutic effect was slower.. Qingyi decoction ameliorates acute pancreatitis-induced intestinal barrier injury by inhibiting the overexpression of intestinal sPLA2. This mechanism may be similar to that of verapamil.

Topics: Acute Disease; Amine Oxidase (Copper-Containing); Amylases; Animals; Anti-Inflammatory Agents; Apoptosis; Deoxycholic Acid; Dexamethasone; Disease Models, Animal; Drugs, Chinese Herbal; Epithelial Cells; Gene Expression Regulation, Enzymologic; Intestinal Mucosa; Male; Pancreatitis; Phospholipases A2, Secretory; Rats, Sprague-Dawley; RNA, Messenger; Severity of Illness Index; Tumor Necrosis Factor-alpha; Verapamil

To investigate the role and potential mechanisms of bone marrow mesenchymal stem cells (MSCs) in severe acute peritonitis (SAP).. Pancreatic acinar cells from Sprague Dawley rats were randomly divided into three groups: non-sodium deoxycholate (SDOC) group (non-SODC group), SDOC group, and a MSCs intervention group (i.e., a co-culture system of MSCs and pancreatic acinar cells + SDOC). The cell survival rate, the concentration of malonaldehyde (MDA), the density of superoxide dismutase (SOD), serum amylase (AMS) secretion rate and lactate dehydrogenase (LDH) leakage rate were detected at various time points. In a separate study, Sprague Dawley rats were randomly divided into either an SAP group or an SAP + MSCs group. Serum AMS, MDA and SOD, interleukin (IL)-6, IL-10, and tumor necrosis factor (TNF)-α levels, intestinal mucosa injury scores and proliferating cells of small intestinal mucosa were measured at various time points after injecting either MSCs or saline into rats. In both studies, the protective effect of MSCs was evaluated.. In vitro, The cell survival rate of pancreatic acinar cells and the density of SOD were significantly reduced, and the concentration of MDA, AMS secretion rate and LDH leakage rate were significantly increased in the SDOC group compared with the MSCs intervention group and the Non-SDOC group at each time point. In vivo, Serum AMS, IL-6, TNF-α and MAD level in the SAP + MSCs group were lower than the SAP group; however serum IL-10 level was higher than the SAP group. Serum SOD level was higher than the SAP group at each time point, whereas a significant between-group difference in SOD level was only noted after 24 h. Intestinal mucosa injury scores was significantly reduced and the proliferating cells of small intestinal mucosa became obvious after injecting MSCs.. MSCs can effectively relieve injury to pancreatic acinar cells and small intestinal epithelium, promote the proliferation of enteric epithelium and repair of the mucosa, attenuate systemic inflammation in rats with SAP.

Topics: Acute Disease; Amylases; Animals; Cell Proliferation; Cell Survival; Cells, Cultured; Coculture Techniques; Deoxycholic Acid; Disease Models, Animal; Inflammation Mediators; Interleukin-10; Interleukin-6; Intestinal Mucosa; L-Lactate Dehydrogenase; Male; Malondialdehyde; Mesenchymal Stem Cell Transplantation; Oxidative Stress; Pancreas, Exocrine; Pancreatitis; Rats; Rats, Sprague-Dawley; Severity of Illness Index; Superoxide Dismutase; Time Factors; Tumor Necrosis Factor-alpha

Triggering receptor expressed on myeloid cells-1 (TREM-1) is a regulator of immunity and an amplifier of inflammatory signaling. The aim was to clarify the role of TREM-1 in the pathophysiology of experimental severe acute pancreatitis (SAP).. SAP was induced by retrograde injection of 3 and 20% sodium deoxycholate (DCA) into the biliopancreatic ducts in rats (DCA pancreatitis). Soluble TREM-1 levels in serum, ascitic fluid, pancreas, liver and kidney were determined with an established available enzyme-linked immunosorbent assay (ELISA) kit. To clarify the source of soluble TREM-1 in serum and ascitic fluid, peritoneal macrophage depletion was done. Moreover, the effect of blockade of TREM-1 pathway was examined using LP17 (a synthetic TREM-1 inhibitor).. Soluble TREM-1 levels in serum and ascitic fluid were higher in SAP. Membrane-bound TREM-1 protein was increased in pancreas, liver and kidney in SAP. Peritoneal macrophage depletion resulted in the reduction of soluble TREM-1 levels in serum and ascitic fluid. Pretreatment with LP17 improved the hepatic and renal dysfunction (serum aspartate aminotransferase and blood urea nitrogen levels) in SAP.. TREM-1 may act as an important mediator for inflammation and organ injury in SAP. TREM-1 may be a potential therapeutic target for the development of SAP and associated organ dysfunction.

Topics: Acute Disease; Animals; Ascitic Fluid; Deoxycholic Acid; Kidney; Liver; Male; Pancreas; Pancreatitis; Rats; Rats, Wistar; Receptors, Immunologic; Triggering Receptor Expressed on Myeloid Cells-1

In severe acute pancreatitis (SAP), multiple organ dysfunction syndrome is a contributor to high mortality. We recently demonstrated that the serum interleukin (IL)-15 level is a predictor of the complications and mortality in clinical SAP. The aim was to investigate the role of IL-15 in experimental SAP.. SAP was induced by retrograde injection of 3 and 20% sodium deoxycholate (DCA) into biliopancreatic ducts in rats (DCA pancreatitis). Expressions of IL-15 were evaluated by Western blotting and immunohistochemical staining. Recombinant IL-15 protein was administered intraperitoneally, and the effects were investigated.. Western blotting revealed the expressions of IL-15 in the pancreas, liver, lung and intestine in 3% DCA pancreatitis. Immunohistochemical staining showed the expression of IL-15 in the cytoplasm of each organ. In 3% DCA pancreatitis, administration of recombinant IL-15 protein attenuated the elevation of serum alanine aminotransferase (ALT) levels and improved the morphological change of the lung 18 h after the induction of SAP. Moreover, in 20% DCA pancreatitis, IL-15 improved the elevation of serum amylase and ALT levels 6 h after the induction.. These results suggest that IL-15 is related to organ dysfunction during SAP, and that IL-15 functions as a protective factor against the organ injuries.

Topics: Alanine Transaminase; Amylases; Animals; Deoxycholic Acid; Humans; Immunohistochemistry; Interleukin-15; Intestine, Small; Lung Injury; Male; Pancreatitis; Rats; Rats, Wistar; Recombinant Proteins; Tissue Distribution

To investigate the effects of serum and ascitic fluid from rats with acute pancreatitis (AP) on cellular free calcium concentration ([Ca(2+)]i) of isolated rat pancreatic acinar cells, and the intervention of pyrrolidine dithiocarbamate (PDTC) and tetrandrine (Tet) to cellular calcium overload in AP.. AP was induced in Sprague-Dawley rats with a retrograde pancreatic duct injection of 3% sodium deoxycholate, and confirmed by histopathological examination and amylase activity assay. The rat serum and ascitic fluid were collected at 1, 5 and 10 h after AP induction, and used as irritants on isolated rat pancreatic acinar cells. The effects on intracellular [Ca(2+)]i, and cell viability were examined. Then, the antagonistic effects of different concentrations of PDTC and Tet were assessed.. The irritation with AP serum and ascitic fluid reduced the survival rate of the isolated rat pancreatic acinar cells and increased the cellular [Ca(2+)]i significantly (P < 0.05). As AP induction course prolonged, the stimulation effect of the AP serum and ascitic fluid intensified. In the pretreated acinar cells with PDTC or Tet, the decreased cell vitality reverted. The elevation of [Ca(2+)]i in the acinar cells significantly ameliorated (significant, P < 0.05; very significant, P < 0.01).. The serum and ascitic fluid from AP rats drastically elevate the [Ca(2+)]i in isolated pancreatic acinar cells and decrease cell vitality, while the pretreatment of cells with PDTC and Tet offsets the calcium overload irritated by the AP serum and ascitic fluid and protects these isolated acinar cells.

Topics: Acute Disease; Amylases; Animals; Anti-Inflammatory Agents; Ascitic Fluid; Benzylisoquinolines; Calcium; Calcium Channel Blockers; Cell Survival; Cells, Cultured; Deoxycholic Acid; Disease Models, Animal; NF-kappa B; Pancreas, Exocrine; Pancreatitis; Pyrrolidines; Rats; Rats, Sprague-Dawley; Thiocarbamates; Time Factors

Bacterial translocation (BT) plays an important role in systemic complications in severe acute pancreatitis (SAP). We recently demonstrated that accelerated apoptosis of intestinal mucosa might have a role in BT. Effects of vascular endothelial growth factor (VEGF) on intestinal epithelial cell apoptosis and BT were investigated in SAP.. Severe acute pancreatitis was induced by retrograde injection of sodium deoxycholate into the biliopancreatic duct in rats. Recombinant rat VEGF (2 microg) was injected, and SAP was immediately induced. Eight hours after the induction, serum amylase/lipase levels and apoptosis of ileal mucosa were evaluated. After 18 hours, the villous height of ileum was examined. After 22 hours, hematocrit, pancreatic water content, BT to the mesenteric lymph nodes, plasma plasminogen activator inhibitor 1 levels, and microvessel density in the small intestine were investigated.. Amylase/lipase levels were significantly elevated in SAP, but VEGF did not affect them. Apoptosis of ileal mucosa was accelerated in SAP, and VEGF significantly reduced the apoptosis. Villous height was significantly decreased in SAP, and VEGF significantly improved it. Vascular endothelial growth factor did not affect the hematocrit or pancreatic water content. Bacterial translocation occurred in the SAP group, and VEGF significantly prevented that. Plasminogen activator inhibitor 1 levels were significantly elevated in SAP, and VEGF significantly improved the elevation. Microvessel counts were significantly reduced in SAP, and VEGF significantly increased them.. These results suggest that VEGF inhibits intestinal epithelial cell apoptosis and subsequent BT in SAP.

Topics: Acute Disease; Angiogenesis Inducing Agents; Animals; Apoptosis; Bacterial Translocation; Deoxycholic Acid; Disease Models, Animal; Gastrointestinal Agents; Intestinal Mucosa; Intestine, Small; Lymph Nodes; Male; Mesentery; Neovascularization, Physiologic; Pancreatitis; Rats; Rats, Wistar; Recombinant Proteins; Severity of Illness Index; Time Factors; Vascular Endothelial Growth Factor A

Topics: Acute Disease; Animals; Ascitic Fluid; Ceruletide; Deoxycholic Acid; Disease Models, Animal; High Mobility Group Proteins; HMGB1 Protein; Intestine, Small; Kidney; Liver; Lung; Male; Pancreas; Pancreatitis; Rats; Rats, Wistar; Repressor Proteins; Severity of Illness Index; Time Factors

To investigate the changes of chaperonin60 (Cpn60) and pancreatic enzymes in pancreatic acinar cells, and to explore their roles in the development of experimental diabetes and acute pancreatitis (AP).. Two different pathological models were replicated in Sprague-Dawley rats: streptozotocin-induced diabetes and sodium deoxycholate-induced AP. The contents of Cpn60 and pancreatic enzymes in different compartments of the acinar cells were measured by quantitative immunocytochemistry.. The levels of Cpn60 significantly increased in diabetes, but decreased in AP, especially in the zymogen granules of the pancreatic acinar cells. The elevation of Cpn60 was accompanied with the increased levels of pancreatic lipase and chymotrypsinogen in diabetes. However, a decreased Cpn60 level was accompanied by high levels of lipase and chymotrypsinogen in AP. The amylase level was markedly reduced in both the pathological conditions.. The equilibrium between Cpn60 and pancreatic enzymes in the acinar cells breaks in AP, and Cpn60 content decreases, suggesting an insufficient chaperone capacity. This may promote the aggregation and autoactivation of the premature enzymes in the pancreatic acinar cells and play roles in the development of AP.

Topics: Amylases; Animals; Chaperonin 60; Chymotrypsinogen; Deoxycholic Acid; Diabetes Mellitus, Experimental; Lipase; Male; Pancreas; Pancreatitis; Rats; Rats, Sprague-Dawley

We previously reported that serum hepatocyte growth factor (HGF) levels are elevated in patients with acute pancreatitis and that pancreatitis-associated ascitic fluid (PAAF) contains cytotoxic factor(s) inducing apoptosis on Madin-Darby canine kidney (MDCK) cells. In this study, plasma HGF levels and HGF tissue distribution were investigated in rats with experimental acute pancreatitis, and the effects of HGF on the cytotoxic activity and apoptosis-inducing activity of PAAF also were examined. Plasma HGF levels were elevated in rats with two experimental pancreatitis models of different grades of severity. The degree of its elevation was correlated with the severity and the organ dysfunctions. In rats with severe pancreatitis, HGF protein and messenger RNA (mRNA) levels significantly increased in liver, kidney, and lung, which were injured organs. When anti-HGF neutralizing antibody was administered in severe pancreatitis, liver dysfunction worsened, and apoptotic cells increased in kidney. Recombinant HGF inhibited the cytocidal activity of PAAF on MDCK cells in a dose-dependent manner. Moreover, recombinant HGF prevented the apoptotic cell death (DNA fragmentation, nuclear fragmentation, and caspase-3 activation) induced by PAAF. These results suggest that HGF is produced in injured organs and may function as an organotrophic and antiapoptotic factor against the organ injuries in acute pancreatitis.

Topics: Acute Disease; Animals; Apoptosis; Caspase 3; Caspases; Cell Line; Ceruletide; Deoxycholic Acid; DNA Fragmentation; Dogs; Edema; Hepatocyte Growth Factor; Kidney; Liver; Lung; Male; Multiple Organ Failure; Pancreatitis; Pancreatitis, Acute Necrotizing; Rats; Rats, Wistar; Recombinant Fusion Proteins; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Spleen

The mechanism of acute pancreatitis-induced hepatocellular injury is unclear. We have observed hepatocyte apoptosis in rat acute necrotizing pancreatitis. These studies were designed to determine the mediator(s) responsible for hepatocyte apoptosis and to clarify the significance of macrophages as its source.. A rat sodium deoxycholate-induced pancreatitis model was used. Immunohistochemical studies for apoptosis-inducing mediators on hepatocytes were examined in the liver and on the peritoneal macrophages. The levels of transforming growth factor-beta1 (TGF-beta1) were also evaluated quantitatively with an enzyme-linked immunosorbent assay. Induction of apoptosis on the hepatocytes was evaluated by in situ nick-end labeling and tissue DNA fragmentation enzyme-linked immunosorbent assay. Finally, the effects of TGF-beta1 neutralization and macrophage depletion were examined.. In the liver and the peritoneal macrophages, strong expression of TGF-beta1 was detected early in the course of pancreatitis. In sodium deoxycholate-induced pancreatitis, the levels of TGF-beta1 were also elevated in the plasma (9.2 +/- 0.8 ng/mL), in the pancreatitis-associated ascitic fluid (11.5 +/- 0.6 ng/mL), and in the liver homogenate (2.8 +/- 0.3 ng/g of liver tissue). Moreover, the amount of fragmented DNA of the liver with pancreatitis was 290% +/- 20% of that with a sham operation and serum alanine aminotransferase levels elevated to 248.2 +/- 67.0 IU/L. TGF-beta1 neutralization partly blocked the positive labeling on the nuclei of the hepatocytes, the elevation of the amounts of fragmented DNA (205% +/- 10% of sham operation), and the serum alanine aminotransferase level (144.2 +/- 14.9 IU/L). On the other hand, the macrophage depletion caused a marked decrease in the TGF-beta1 protein level in the plasma (4.8 +/- 1.2 ng/mL) or in the pancreatitis-associated ascitic fluid (8.0 +/- 1.0 ng/mL). Moreover, the macrophage depletion completely inhibited the elevation of the TGF-beta1 protein level in the liver homogenate (1.5 +/- 0.4 ng/g of liver tissue), and thereafter decreased the amounts of the positive labeling on the nuclei of the hepatocytes and decreased the amount of fragmented DNA (120% +/- 18% of sham operation) and the serum alanine aminotransferase elevation (119.2 +/- 24.2 IU/L).. In a model of sodium deoxycholate-induced pancreatitis, macrophages are responsible for pancreatitis-induced hepatocellular injury by means of apoptosis, and macrophage-derived TGF-beta1 is one of the major factors inducing the hepatocyte apoptosis.

Topics: Acute Disease; Animals; Apoptosis; Clodronic Acid; Deoxycholic Acid; Disease Models, Animal; DNA Fragmentation; Immunohistochemistry; Kupffer Cells; Liposomes; Liver; Macrophages, Peritoneal; Male; Neutralization Tests; Pancreatitis; Rats; Rats, Wistar; Transforming Growth Factor beta

Bacterial translocation leading to subsequent infectious complications is a significant determinant of outcome in acute hemorrhagic pancreatitis (AHP). The colonic ileus and impaired intestinal barrier function that often accompany AHP may predispose to translocation. Sennoside is a naturally occurring cathartic and choleretic agent that stimulates intestinal mucous secretion and has potent promotility effects. The impact of sennoside-induced intestinal motility and secretory function on bacterial translocation and survival was studied in a rat model of AHP. Severe acute pancreatitis was induced in rats by the intraductal infusion of 2% sodium deoxycholate (DCA, 0.4 ml/kg). A group of sham-operated rats (group A) received intraductal saline, whereas experimental animals were subsequently administered distilled water (group B) or sennoside solution (group C) by gavage every 8 h. After 48 h, intestinal transit of fluorescein isothiocyanate-labeled dextran, serum endotoxin, and amylase levels, and bacterial translocation to mesenteric lymph nodes (MLNs) and pancreatic tissue were determined. The pancreas and intestine were sampled for histologic study. All group A animals survived and did not develop pancreatitis or endotoxemia, whereas groups B and C all demonstrated severe hemorrhagic pancreatitis with evidence of necrosis. Mortality at 48 h was 55% in group B versus 12.5% in group C. Inhibition of intestinal motility was noted in 40% versus 20%, and endotoxin levels were 61.36+/-28.26 pg/L versus 5.41+/-3.58 pg/L in group B versus group C rats, respectively (p<0.001). Pancreatic tissue and MLN cultures were positive in 100% of group B survivors versus 14% of group C survivors (p<0.05). Histologic examination of the intestine in group C animals showed increased mucous secretion, proliferation of goblet cells, and evidence of rapid turnover/renewal of enterocytes. Treatment with the cathartic agent, sennoside, reduced translocation of endotoxin and bacteria, restored intestinal motility, increased mucous secretion, and reduced mortality in a model of acute hemorrhagic pancreatitis in the rat. Other cathartics may have similar properties and may be useful in preventing infectious complications in acute pancreatitis.

Topics: Acute Disease; Amylases; Animals; Anthraquinones; Bacterial Infections; Cathartics; Deoxycholic Acid; Disease Models, Animal; Endotoxemia; Endotoxins; Gastrointestinal Motility; Hemorrhage; Intestinal Mucosa; Jejunum; Lymph Nodes; Male; Necrosis; Pancreatic Diseases; Pancreatitis; Rats; Rats, Wistar; Senna Extract; Sennosides

Multiple organ failure (MOF) is the most serious complication in severe acute pancreatitis, contributing to its high mortality. It has been suggested that changes of high-energy phosphates, intracellular pH, and intracellular cation homeostasis are closely related to hepatocellular injury associated with MOF.. Phosphorus metabolites, intracellular pH (pHi), and intracellular Na+ concentration ([Na+]i) were measured in rat livers in vivo using 31P and 23Na NMR spectroscopy after deoxycholic acid (DCA)-induced pancreatitis or intraperitoneal injection (ip) of pancreatitis-induced ascitic fluid (PAF).. Two hours after induction of DCA-pancreatitis, the liver experienced significant intracellular acidosis (pHi = 6.99 +/- 0.16) and sodium loading (75 +/- 9 mM) and a reduction in its energy state (beta-ATP/Pi = 0.2 +/- 0.03 and Pi = 164 +/- 12). Although ip injection of PAF into healthy rats did not induce systemic hypotension, the livers under these conditions also developed severe disturbances in hepatocellular ion homeostasis and depletion of its bioenergetics. The longer the abdomen was exposed to the PAF, the worse the changes were. At 3 h after ip injection of PAF, hepatic [Na+]i significantly increased (42 +/- 3 mM) along with a significant decrease in pHi (7.30 +/- 0. 03). At 6 h after ip injection of PAF, the hepatic beta-ATP/Pi ratio decreased to 0.34 +/- 0.05 and Pi increased to 97 +/- 27.. PAF induced severe hepatocellular acidosis, rapid accumulation of hepatic intracellular sodium, impaired hepatic cytosolic phosphorylation potential, and increased hepatic utilization of ATP. These effects may account for the eventual development of liver dysfunction associated with necrotizing pancreatitis.

Topics: Acute Disease; Animals; Ascitic Fluid; Deoxycholic Acid; Energy Metabolism; Hydrogen-Ion Concentration; Intracellular Membranes; Liver; Magnetic Resonance Spectroscopy; Male; Pancreatitis; Phosphorus; Rats; Rats, Sprague-Dawley; Sodium

Two different types of secretory phospholipase A2 (PLA2), pancreatic group I (PLA2-I) and non-pancreatic group II (PLA2-II), have been identified and postulated to be associated with the pathogenesis of various diseases, such as acute pancreatitis, septic shock, and multiple organ failure.. To investigate the type of secretory PLA2 responsible for its catalytic activity found in plasma and ascites of experimental acute pancreatitis.. Acute pancreatitis of differing severity was induced by the injection of different concentrations (1% or 10%) of sodium deoxycholate (DCA) into the common biliopancreatic duct in rats, and catalytic PLA2 activity in plasma and ascites were differentiated by anti-PLA2-I antibody and specific inhibitor of PLA2-II. Survival rate and plasma amylase, aspartate aminotransferase (AST), and alanine aminotransferase (ALT) were also measured.. In 1% and 10% DCA induced acute pancreatitis, plasma amylase values as well as PLA2 activity in ascites were greatly increased. PLA2 activity in plasma was also notably increased in 10% DCA induced acute pancreatitis, but not in 1% DCA induced acute pancreatitis. PLA2-I specific polyclonal antibody significantly inhibited PLA2 activity in ascites but not that in plasma. In contrast, plasma PLA2 activity was completely suppressed by PLA2-II specific inhibitor. In addition, a high mortality (93% at five hours) and a significant increase in plasma AST and ALT were noted in 10% DCA induced pancreatitis.. Ascites PLA2 activity is mainly derived from PLA2-I, whereas plasma PLA2 activity is mostly derived from PLA2-II in severe acute pancreatitis, suggesting that increased plasma PLA2-II activity might be implicated in hepatic failure arising after severe acute pancreatitis.

Topics: Acute Disease; Alanine Transaminase; Amylases; Animals; Aspartate Aminotransferases; Biological Assay; Cholagogues and Choleretics; Deoxycholic Acid; Group II Phospholipases A2; Liver Failure; Male; Pancreatitis; Phospholipases A; Phospholipases A2; Rats; Rats, Wistar; Survival Rate

The acute pancreatitis (AP) was induced with sodium deoxy-cholate in SD rats. Succinic dehydrogenase (SDH), acid phosphatase (ACP) and cytochrome P450 were taken as marked enzyme in estimation of mitochondria, lysosome and microsome to observe the functional and structural changes of the pancreatic subcellular organs and the protective effects of Fructus Gardeniae (FG) in acute pancreatitis in rats. The histological change was observed simultaneously. The results showed that pancreatic cellular SDH reduced significantly and the releasing rate of ACP raised obviously in AP, in FG treated group, SDH was nearly the same as in normal, the releasing rate of ACP decreased and the content of P450 increased in the same time. Histologic observation showed that there was significant inflammatory change in pancreas and abnormal manifestation in structure of subcellular organs in AP, FG Could alleviate the lesion. These results suggested the FG has a remarkable protective effect on the function and structure of the pancreatic subcellular organs in AP.

Topics: Acute Disease; Animals; Deoxycholic Acid; Drugs, Chinese Herbal; Female; Male; Pancreas; Pancreatitis; Rats; Rats, Sprague-Dawley; Succinate Dehydrogenase

Interleukin-1 (IL-1) is a mediator in some critical conditions such as septic shock and multiple organ failure. Acute pancreatitis is one of the noted causes of multiple organ failure but the mechanism by which local inflammation progresses to systemic disease is unknown. In this study, we used an IL-1 receptor antagonist (IL-1ra) to investigate whether multiple organ failure due to acute pancreatitis is mediated by IL-1, as in other causes such as severe infection, trauma, and major surgery.. Prospective, randomized, controlled trial.. Research laboratory of a university medical school.. Specific pathogen-free male Wistar rats weighing 200 to 250 g.. Necrotizing pancreatitis was induced by retrograde injection of deoxycholate solution into the biliopancreatic duct. IL-1ra was injected intravenously at a dose of 10 mg/kg 15 mins before induction of acute pancreatitis and then infused continuously at a rate of 5 mg/kg/hr for the following 24 hrs.. Although treatment with recombinant human IL-1ra did not affect the degree of local pancreatic insult, it significantly reduced mortality, improved urine output as an indicator of the state of shock, and ameliorated the accumulation of neutrophils into the lung in a rat experimental pancreatitis model.. We concluded that multiple organ failure in severe pancreatitis is mediated, at least in part, by IL-1 through the activation of neutrophils. Furthermore, we concluded that circulatory collapse may also be important in the mechanism of the lethal effect of pancreatitis.

Topics: Acute Disease; Animals; Deoxycholic Acid; Disease Models, Animal; Drug Evaluation, Preclinical; Interleukin 1 Receptor Antagonist Protein; Interleukin-1; Male; Multiple Organ Failure; Necrosis; Pancreas; Pancreatitis; Prospective Studies; Random Allocation; Rats; Rats, Wistar; Receptors, Interleukin-1; Recombinant Proteins; Sialoglycoproteins; Specific Pathogen-Free Organisms

The integrity of rat pancreatic acinar cells under the influence of human phospholipase A2 (PLA2) was studied. Isolated pancreatic acini showed no increased discharge of aspartylaminotransferase (ASAT) when incubated either in solutions containing human pancreatic PLA2 or the bile salt sodium deoxycholate (DEC), the latter in concentrations that augment PLA2 activity but have no destructive detergent effect. When human pancreatic PLA2 was injected into the rat pancreatic duct, uneven distribution was observed at 15 min and 3 h in immunohistochemical sections. Edema and a mild inflammatory reaction were the main changes in the pancreas. The necrotic areas seen by light and electron microscopy were quite small and located mostly at the periphery of lobules corresponding the spread of the injected material. Necrosis was of the coagulation type and showed equal extent after the injection of PLA2 with or without DEC. Internalized human pancreatic PLA2 was present already 15 min after the injection in the cytoplasm of some intact acinar cells, indicating a functioning protective mechanism. It was concluded that pancreatic acinar cells are quite resistant to PLA2-catalyzed hydrolysis of membrane phospholipids in vitro, but additional trauma, e.g., pressure caused by intraductal injection, and tissue related factors, such as the mediators of the inflammatory reaction, make acinar cells susceptible to the effect of PLA2.

Topics: Acute Disease; Animals; Aspartate Aminotransferases; Deoxycholic Acid; In Vitro Techniques; Male; Necrosis; Pancreas; Pancreatitis; Phospholipases A; Phospholipases A2; Rats; Rats, Inbred Strains

We describe an experimental model of acute pancreatitis in conscious rats, based on biliary reflux; sodium deoxycholate was injected while maintaining the common duct (biliary and pancreatic) temporally occluded. A progressive increase of serum amylase, cardiac rate and hematocrit value, as well as a decrease of plasma proteins are characteristic of this experimental model. Blood pressure was maintained until shortly before death, which occurred after 85 minutes. This is a simple model, reliable and reproducible, which requires minimal surgical manipulation.

Topics: Acute Disease; Animals; Common Bile Duct; Deoxycholic Acid; Disease Models, Animal; Male; Pancreatitis; Rats; Rats, Inbred Strains

Therapeutic use of the perfluorochemical emulsion, Fluosol DA, in acute pancreatitis was experimentally discussed in view of maintaining the local blood flow and oxygen supply in the pancreas to avoid further aggravation of pancreatitis. Acute pancreatitis was induced by deoxycholate injection into the pancreatic duct in adult mongrel dogs. Fluosol DA or 6% hydroxyethylstarch (HES) solution as control was transfused at 20 ml/kg/hour for the first 3 hours. Fluosol DA and HES solution improved the depressed cardiac output and pancreatic blood flow to normal levels. Compared with HES solution, Fluosol DA administration revealed a prominent increase in oxygen tension in the pancreatic tissue, which had decreased severely from onset of pancreatitis. Fluosol DA administration brought about better preservation of pancreatic mitochondrial functions. Despite no significant differences in blood levels of other pancreatic enzymes between Fluosol DA and HES solution, the sharp decrease in plasma postheparin phospholipase A2 suggested the protection of involved systemic organs including pancreas. Thus, maintaining pancreatic blood flow and increasing the oxygen transport by Fluosol DA administration seemed to play a positive role in inhibiting the progress of pancreatitis, though improvement of survival rate in acute pancreatitis was incomplete by Fluosol DA administration alone.

Topics: Acute Disease; Animals; Blood Substitutes; Cardiac Output; Deoxycholic Acid; Dogs; Drug Combinations; Fluorocarbons; Hydroxyethyl Starch Derivatives; Kinetics; Oxygen; Pancreas; Pancreatitis; Phospholipases A; Phospholipases A2; Regional Blood Flow

Serum lipase levels are much greater in cases of chronic alcoholics (without any marked symptom of pancreatitis) than in healthy subjects. In fact, about 43 p. cent of the studied samples from alcoholics exhibit a lipase activity above the reference interval (0-160 U/l). On the other hand, the lipase activity present in the serum of alcoholics exhibits different properties than lipoprotein lipase or post-heparin plasma lipase. Furthermore the enzyme from alcoholics presents similar properties to those of the serum lipase released in cases of pancreatitis mainly concerning the sensitivity versus colipase and biliary salt concentration. This similarity with the "pancreatitis" enzyme suggests a pancreatic disorder in number of cases of chronic alcoholics. Therefore, the authors think that serum lipase activity could be taken into account in the evaluation of the risk of any abnormality in the pancreatic function in all alcoholics.

Topics: Acute Disease; Alcoholism; Colipases; Deoxycholic Acid; Enzyme Activation; Humans; Lipase; Pancreatitis

Small amounts (0.1-0.5 mM) of deoxycholate enhanced amylase secretion, which had been induced by submaximal doses of carbachol or cholecystokinin octapeptide, without affecting the maximal levels of these reactions from isolated rat pancreatic acini. Deoxycholate alone did not induce these reactions. The other bile acids such as cholate, chenodeoxycholate, ursodeoxycholate, and taurocholate were also active. Under the similar conditions, deoxycholate enhanced the secretagogue-induced diacylglycerol formation that was derived mainly from the phospholipase C-mediated hydrolysis of phosphatidylinositol and phosphatidylinositol-4-monophosphate. Deoxycholate did not enhance the secretagogue-induced hydrolysis of phosphatidylinositol-4,5-bisphosphate or Ca2+ mobilization. Deoxycholate did not affect amylase secretion, which was induced by the simultaneous addition of protein kinase C-activating 12-O-tetradecanoylphorbol-13-acetate and Ca2+ ionophore ionomycin. Since diacylglycerol and Ca2+ may be responsible for the secretagogue-induced amylase secretion, our results indicate that small amounts of bile acids increase the sensitivity to the secretagogue of diacylglycerol formation and subsequent activation of protein kinase C, and thereby enhance amylase secretion from pancreatic acini.

Topics: Amylases; Animals; Bile Acids and Salts; Calcium; Deoxycholic Acid; Diglycerides; Dose-Response Relationship, Drug; In Vitro Techniques; Male; N-Methylscopolamine; Octoxynol; Pancreas; Pancreatitis; Phosphatidylinositols; Polyethylene Glycols; Rats; Rats, Inbred Strains; Scopolamine Derivatives; Sincalide; Tetradecanoylphorbol Acetate

A photometric method for the determination of phospholipase A is described, in which fatty acids are liberated from phosphatidylcholine and measured by a discontinuous enzymatic test. Optimal test conditions were evaluated using purified phospholipase A from human and porcine pancreas. The enzyme activities measured in normal human sera are in the range of 0 to 10 U/l. Pathological values reach more than 100 U/l. There is no correlation with the activities of lipase or amylase in cases of acute pancreatitis.

Topics: Amylases; Animals; Deoxycholic Acid; Humans; Hydrogen-Ion Concentration; Kinetics; Lipase; Pancreas; Pancreatitis; Phospholipases; Phospholipases A; Photometry; Swine

Acute pancreatitis can be induced experimentally in rats by the retrograde pancreatic duct injection of deoxycholic acid. In order to evaluate if prostaglandins have a protective or therapeutic role, pancreatitis was induced with 0.03, 0.1, or 0.2 ml of 3% deoxycholic acid. Then, 16,16 dimethyl prostaglandin E2 or prostaglandin I2 were infused i.v. for 30 min before the deoxycholic acid injection or for 1 or 24 h beginning 30 min or 1 h after the injection. Deoxycholic acid, 0.1 and 0.2 ml, produced an 80-100% mortality at 24 h after the induction of pancreatitis, and prostaglandin administration had no appreciable effect. Induction of pancreatitis with 0.03 ml deoxycholic acid was associated with a 15% mortality at 24 h, and both prostaglandins significantly increased the mortality. In the experimental model of pancreatitis tested, the administration of exogenous PGI2 and 16,16 dimethyl PGE2 significantly worsened the outcome.

Topics: Acute Disease; Animals; Deoxycholic Acid; Epoprostenol; Female; Pancreatitis; Prostaglandins E, Synthetic; Rats; Rats, Inbred Strains

Topics: Acute Disease; Adult; Aged; Deoxycholic Acid; Female; Humans; Hyperlipidemias; Male; Middle Aged; Pancreatitis; Recurrence; Ursodeoxycholic Acid

The rate of protein phosphorylation, as catalyzed by the protein kinase enzymes, was measured in the pancreas of rats with acute experimental pancreatitis. Two different methods were used to induce pancreatitis in rats: retrograde injection of deoxycholate (DOC) into the pancreatic duct, or daily intravenous administration of DL-ethionine. Basal protein kinase activity was elevated in rats with acute experimental pancreatitis. This increase in activity was not dependent on free Ca2+ and did not result from elevated cAMP levels. To assess the possible role of digestive enzymes in protein kinase activation, tissue extracts from healthy controls were subjected to mild treatment with digestive enzymes and DOC. Trypsin, chymotrypsin, phospholipase A, and DOC produced protein kinase activation of a similar magnitude as found in diseased tissue. Results indicate that stimulated protein kinase activity in tissue of animals with acute pancreatitis may arise from the action of digestive enzymes.

Topics: Acute Disease; Animals; Bile Acids and Salts; Carboxypeptidases; Chymotrypsin; Deoxycholic Acid; Ethanol; Lipolysis; Pancreas; Pancreatitis; Phospholipases; Phosphorylation; Protein Kinases; Rats; Rats, Inbred Strains; Trypsin

In the last years non operative and surgical efforts could not diminish the high lethality of severe acute haemorrhagic necrotising pancreatitis. While majority of patient die in consequence of pancreatitis-shock, hyperbaric oxygen therapy (HBO) can improve all hypoxic circulation situations. Therefore the value in treatment of experimental necrotising pancreatitis in pig is examined. 5 pigs were treated with, 5 without HBO and 5 served as control. With HBO liquid sequestration was diminished and total protein loss prevented significantly, but foremost survival time was significantly prolonged. In consequence demarcation of necrosis with connective tissue was possible, but nevertheless operative treatment remain necessary. Without HBO all pigs and with HBO only 2 pigs died in consequence of necrotising pancreatitis. It is demonstrated, that HBO as additive therapy can improve the prognosis of necrotising pancreatitis.

Topics: Acute Disease; Animals; Deoxycholic Acid; Female; Hyperbaric Oxygenation; Male; Necrosis; Pancreatitis; Prognosis; Swine

Topics: Amylases; Animals; Aspirin; Body Weight; Deoxycholic Acid; Dietary Fats; Female; Indomethacin; Pancreatitis; Rats; Sodium Chloride

We propose a rapid enzymatic micromethod for the specific determination of lipase (EC 3.1.1.3) activity in serum and duodenal fluid. Free linoleic acid produced during 10-min incubation of 10 mul of sample with 1 ml of substrate (trillinolein emulsion) at 30 degrees C is converted by lipoxygenase (EC 1.99.2.1), in a coupled reaction, to its hydroperoxide, which is measured photometrically after solubilizing the reaction mixture in ethanol. Lipase activity is calculated from the rate of hydroperoxide formation, with linoleic acid as primary standard. The velocity of the reaction is greatest at pH 8.8, 35-37 degrees C, and a deoxycholate concentration of 3.6 mmol/liter. The energy of activation is 6.7 kcal/mol. The differing "apparent" Km values obtained for lipase in undiluted serum (4 X 10(-5) mol/liter) and in albumin-based diluents (1 X 10(-5) mol/liter) indicate the presence of a competitive inhibitor in the serum matrix. We detected no lipase activity in urine. Results by the proposed method correlate well with those by a copper soap extraction method (r = 0.95), but values are significantly higher for pancreatitis patients' sera (slope 1.6). The linear dynamic range extends to 1000 U/liter. Hemolysis, lipemia, and hyperbilirubinemia do not interfere. The normal range is 40-60 U/liter. Lipase activity of pancreatitis patients generally exceed 1000 U/liter during the acute phase and 250 U/liter for as long as 10 days after it.

Topics: Amylases; Bicarbonates; Deoxycholic Acid; Duodenum; Humans; Hydrogen-Ion Concentration; Kinetics; Linoleic Acids; Lipase; Lipoxygenase; Pancreatitis; Secretin; Temperature

The role of the complement system in the initial membrane lesion of acute pancreatitis was investigated. In the experimental sodium-taurocholate pancreatitis of the rat a sudden and steady decline of serum complement was observed. The deposition of C3 component of complement in acute pancreatitis could be demonstrated by immunofluorescence. To rule out mere deposition or activation of complement in the interstitial exsudative fluid, single acinar cells of rat and rabbit pancreatic tissue were prepared and transfered to culture medium. In contrast to heat inactivated serum and C6 deficient serum these cells were lysed by trypsin activated fresh serum. Consequently, an acute pancreatitis could be induced by activating exclusively the complement system by injection of cobra venom factor into the pancreatic duct of rats. The activated complement system is thought to be responsible for initial membrane lesion in exsudative inflammation, as could be shown in acute pancreatitis.

Topics: Acute Disease; Animals; Complement System Proteins; Deoxycholic Acid; Male; Pancreatitis; Rats; Snakes; Time Factors; Venoms

The proteolytic activities and trypsin inhibitors of the peritoneal exudate produced by experimental acute pancreatitis in the rat were studied by fractionation with gel filtration on Sephadex G-200 and estimation of the hydrolysis of casein and synthetic substrates. The peritoneal exudate produced by injecting formalin solution into the peritoneal cavity was used as a control. The peritoneal exudate during pancreatitis revealed distinct proteolytic and ATEE hydrolysing activities and it also hydrolysed BAPNA to a lesser extent. These activities were absent from the control exudates, or only traces of them could be demonstrated with the methods used. The trypsin inhibiting capacity (TIC) in the pancreatic exudate was about half that in the control exudate. In gel filtration on Sephadex G-200 the BAPNA hydrolysing proteolytic activity was eluted with the macroprotein fraction, suggesting that the enzyme was bound to the macroproteins. TIC differed clearly in the control exudate and the pancreatitis exudate. In both of them TIC was eluted in two peaks after the macroproteins, but in the pancreatitis group the first peak was very weak, if demonstrable at all, while in the control exudate the two peaks were clearly separated and the TIC was more pronounced. These findings suggest that pancreatic enzymes are released during pancreatitis into the peritoneal cavity, where they combine with proteinase binding factors in the exudate.

Topics: Acute Disease; Animals; Ascitic Fluid; Deoxycholic Acid; Male; Pancreatitis; Peptide Hydrolases; Rats; Trypsin Inhibitors

After intraperitoneal application of desoxycholate in cells of exocrine pancreas, ultrastructural alterations are observed. Initial changes are seen in mitochondria and focal parts of the endoplasmic reticulum. In later experimental stages there is sizable increase in the extent of the initially focal degradations, in such a way that great areas of the cytoplasm are involved. It is remarkable to note, that there are no indications about any disturbance in the synthesis and excretion of zymogen granules. The described morphological alterations will be discussed as conceivably being caused primarily by an irritation in the lipoid metabolism of the membrane.

Topics: Animals; Cytoplasmic Granules; Deoxycholic Acid; Endoplasmic Reticulum; Male; Membranes; Mitochondria; Pancreas; Pancreatitis; Rats