ceruletide and Hemorrhage

Octreotide is considered to reduce exocrine pancreatic secretion in acute hemorrhagic necrotizing pancreatitis decreasing pancreatic autodigestion. The aim of this study was to determine whether octreotide also has antioxidative effects in acute pancreatitis. Additionally time and dose of application were of interest.. Ninety male Sprague-Dawley rats were randomized into six groups (n = 15). Group 1 underwent a laparotomy, and animals in groups 2-6 received intraductal glycodeoxycholic acid followed by intravenous cerulein. Groups 3 and 4 were injected with 0.5 mg octreotide, while groups 5 and 6 received continuous intravenous infusion of 0.05 mg octreotide/h for 10 h. Treatment was initiated 6 hours after induction of pancreatitis (IP) in groups 3 and 5, and 14 h after IP in groups 4 and 6. At 24 h after IP all animals were killed and each pancreas was analyzed histopathologically. In addition, levels of pancreatic lipid peroxidation protective enzymes glutathione-peroxidase (GSH-Px) and superoxide dismutase (SOD) as well as lipid peroxidation via thiobarbituric acid reactive substances (TBARS) were determined.. Early bolus application of octreotide reduced severity of histopathological changes in acute pancreatitis and decreased lipid peroxidation in pancreatic tissue samples; however, late bolus application and continuous intravenous infusion did not influence pancreatitis or lipid peroxidation.. Octreotide seems to have a dose- and time-dependent effect on histopathology and lipid peroxidation in a model of pancreatitis in rats.

Topics: Animals; Antioxidants; Ceruletide; Disease Models, Animal; Dose-Response Relationship, Drug; Glutathione Peroxidase; Glycodeoxycholic Acid; Hemorrhage; Infusions, Intravenous; Injections, Intravenous; Lipid Peroxidation; Male; Octreotide; Pancreas; Pancreatitis, Acute Necrotizing; Rats; Rats, Sprague-Dawley; Superoxide Dismutase; Thiobarbituric Acid Reactive Substances; Time Factors

Pancreatic duct obstruction induces edematous but not hemorrhagic pancreatitis even when combined with maximal secretory stimulation. The aim of the present study was to test the hypothesis that pancreatic and bile duct obstruction exacerbates edematous pancreatitis induced by supramaximal secretory stimulation by caerulein.. In in vivo studies using rats, biliopancreatic duct ligation was combined with supramaximal stimulation of caerulein, and pancreatic histology, serum amylase level, pancreatic edema, and intrapancreatic trypsin activation were evaluated. In in vitro studies, the pancreatic acini were isolated from the rats with biliopancreatic duct ligation, and amylase secretion, intracellular trypsin activation, and acinar cell fragility were evaluated.. Biliopancreatic duct ligation exacerbated caerulein-induced pancreatitis from edematous to hemorrhagic only when the obstruction preceded caerulein administration. The amylase secretion from the acini was inhibited, and intracellular trypsin activation and the acinar cell fragility on the supramaximal stimulation with cholecystokinin in vitro were enhanced by the preceding in vivo biliopancreatic duct obstruction.. Preceding biliopancreatic duct obstruction exacerbates caerulein-induced pancreatitis. Enhancement of intracellular trypsin activation is possibly involved in this mechanism.

Topics: Amylases; Animals; Ceruletide; Cholecystokinin; Cholestasis; Constriction, Pathologic; Disease Models, Animal; Disease Progression; Hemorrhage; In Vitro Techniques; Male; Pancreas; Pancreatic Ducts; Pancreatitis; Rats; Rats, Wistar; Trypsin

alpha 1-Acid glycoprotein (AAG), a highly negatively charged glycoprotein, well known for its capillary stabilizing effect, was tested in rat models of acute edematous pancreatitis, acute hemorrhagic-necrotizing pancreatitis, and acute respiratory distress syndrome (ARDS). In cerulein-elicited edematous pancreatitis AAG improved histological alterations at 200 mg/kg i.v. and plasma amylase activity at 1800 or 4200 mg/kg i.v. All other parameters (edema, plasma lipase) were not affected in a biologically relevant manner. In glycodeoxycholic acid-induced hemorrhagic-necrotizing pancreatitis AAG was without effect on parameters measured (plasma amylase, plasma lipase activity, histological scores) at 1800 or 4200 mg/kg i.v. At the extremely high dose of 1500 mg/kg i.v. plasma amylase and lipase levels were decreased. In lipopolysaccharide-mediated ARDS, AAG was tested at 50, 200 or 600 mg/kg i.v. AAG, but also the placebo formulation decreased the myeloperoxidase content in the bronchoalveolar lavage fluid. Histological alterations were improved by AAG, however, not by the placebo formulation. Lung water content was not significantly influenced by AAG, whereas Evans blue extravasation was significantly diminished by all three doses of AAG. It is concluded that the edematous pancreatitis is the first in vivo condition with increased extravascular fluid accumulation, in which AAG is not effective. Based on data presented here and literature data, there is evidence for a beneficial effect of AAG in acute lung injury.

Topics: Acute Disease; Animals; Bronchoalveolar Lavage Fluid; Ceruletide; Edema; Glycodeoxycholic Acid; Hemorrhage; Lipopolysaccharides; Lung Diseases; Male; Orosomucoid; Pancreatitis; Rats; Rats, Sprague-Dawley; Respiratory Tract Diseases

The therapeutic effects of an intravenously injected carboxamide derivative (IS-741) on lung injury were studied in rats with cerulein-induced pancreatitis complicated by endotoxemia. Pancreatitis was induced by four intramuscular injections of cerulein (50 microg/kg at 1-hr intervals). Pancreatitis rats were injected intraperitoneally with 10 mg/kg of lipopolysaccharide (LPS) 6 hr following the first cerulein injection as a challenge of endotoxemia. Rats were divided into four groups: group I, pancreatitis with LPS; group II, pancreatitis with LPS treated with a continuous intravenous injection of IS-741 at 0.03 mg/kg/hr); group III, pancreatitis with LPS treated with a continuous intravenous injection of IS-741 at 0.3 mg/kg/hr); and group IV, pancreatitis with LPS treated with a continuous intravenous injection of IS-741 at 3 mg/kg/hr). IS-741 was administered 30 min before the endotoxemia challenge. Intense mononuclear cell infiltration and lung hemorrhage occurred in untreated pancreatitis rats with LPS (group I), but hemorrhage was not seen in group IV rats receiving a continuous injection of IS-741 shortly before the induction of endotoxemia. The IS-741-treated rats (groups II, III, and IV) had lower serum concentrations of cytokine-induced neutrophil chemoattractant (CINC), as well as fewer pulmonary infiltrates immunoreactive for CINC or Mac-1 (CD11b/CD18). The number of neutrophils infiltrating the lung in groups II, III, and IV was significantly lower than that of group I. Conversely, CINC production by bronchoalveolar macrophages in vitro were stimulated by LPS but were reduced by the presence of IS-741. The carboxamide derivative IS-741 effectively prevented pancreatitis-associated lung injury following the challenge of endotoxemia.

Topics: Acute Disease; Animals; Bronchoalveolar Lavage Fluid; Cells, Cultured; Ceruletide; Endotoxemia; Enzyme Inhibitors; Hemorrhage; Infusions, Intravenous; Interleukin-16; Lung; Lung Diseases; Macrophages; Male; Neutrophils; Pancreatitis; Phospholipases A; Pyridines; Rats; Rats, Sprague-Dawley

We investigated the role of platelet-activating factor (PAF) as a priming signal for cytokine-induced neutrophil chemoattractant (CINC) expression by bronchoalveolar macrophages in acute pancreatitis. Pancreatitis was induced by four intramuscular injections of cerulein (50 micrograms/kg at 1-h intervals) in Wistar rats. The animals were injected intraperitoneally with 10 micrograms/kg of lipopolysaccharide (LPS) as a septic challenge. Pancreatitis rats were treated with a bolus intravenous injection of TCV-309 (3 or 30 micrograms/kg) 30 min before the septic challenge. Intense mononuclear cell infiltration and lung hemorrhage occurred in pancreatitis rats complicated with sepsis but were not seen in pancreatitis rats receiving a bolus TCV-309. Pancreatitis rats treated with TCV-309 had lower serum concentrations of CINC after septic challenge and lower levels of CINC messenger RNA (mRNA) in the lung, as well as fewer pulmonary infiltrates immunoreactive for CINC or Mac-1 (CD11b/CD18). In vitro CINC production in response to LPS by bronchoalveolar macrophages obtained from pancreatitis rats 6 h after the first cerulein injection, immediately before septic challenge, was enhanced but was significantly reduced in a TCV-309-sensitive manner. LPS-stimulated in vitro CINC production by naive bronchoalveolar macrophages was significantly enhanced by pretreatment with PAF. TMB-8 (an inhibitor of calcium release from endoplasmic reticulum) or W7 (calmodulin antagonist) completely abrogated the chemoattractant production by bronchoalveolar macrophages pretreated with PAF after LPS stimulation. Altered intracellular calcium, due to Ca2+ efflux from intracellular stores, may be involved in the "priming" of bronchoalveolar macrophages to release CINC after triggering with LPS during acute cerulein-induced pancreatitis. The PAF antagonist TCV-309 effectively prevented hyperactivity of bronchoalveolar macrophages and pancreatitis-associated lung injury after the septic challenge.

Topics: Animals; Ceruletide; Chemokines, CXC; Chemotactic Factors; Growth Inhibitors; Growth Substances; Hemorrhage; Intercellular Signaling Peptides and Proteins; Isoquinolines; Kinetics; Lipopolysaccharides; Lung; Lung Diseases; Macrophages, Alveolar; Male; Pancreatitis; Platelet Activating Factor; Platelet Aggregation Inhibitors; Pyridinium Compounds; Rats; Rats, Wistar; RNA, Messenger; Sepsis; Tetrahydroisoquinolines

The effect of B2 receptor bradykinin antagonist icatibant on postcapillary leukostasis, microcirculatory stasis, and tissue necrosis was studied in acute pancreatitis. In rats, pancreatitis was induced by intraductal injection of sodium taurocholate (ST), intravenous caerulein and intraductal infusion of glucodeoxycholic acid (GDOC), or intravenous caerulein infusion alone. Intravital pancreatic microcirculation was observed. Icatibant or vehicle was given 30 min before induction of pancreatitis. In ST pancreatitis, the number of perfused capillaries increased in icatibant-pretreated rats (77% vs. 0% for controls, P < 0.001). Capillary flow was preserved in icatibant-treated rats; total stasis was observed in controls. Mean venular leukocyte adherence decreased in icatibant-treated rats (26% vs. 74% for controls, P < 0.001), and median histopathologic score was reduced (icatibant vs. controls, 5.0 vs. 12 points, respectively; P < 0.01). Kinase II inhibitor captopril or exogenous bradykinin in addition to an otherwise effective dosage of icatibant resulted in microcirculatory stasis, extensive venular leukocyte adherence, and severe histological damage. With a 100 times greater icatibant dosage, this adverse effect was compensated. The beneficial effects of icatibant were also observed in intermediate pancreatitis (caerulein + GDOC). In ST and intermediate pancreatitis, icatibant preserved microcirculation, reduced venular leukocyte adherence, and prevented pancreatic tissue damage. B2 receptor bradykinin-mediated postcapillary leukostasis plays an important role in the pathogenesis of severe forms of acute pancreatitis.

Topics: Acute Disease; Animals; Arterioles; Bradykinin; Bradykinin Receptor Antagonists; Capillaries; Cell Adhesion; Ceruletide; Edema; Female; Hemorrhage; Leukocytes; Microcirculation; Pancreas; Pancreatitis; Rats; Rats, Inbred Lew; Receptor, Bradykinin B2; Regional Blood Flow; Taurocholic Acid; Time Factors; Vasoconstriction; Venules

One of the vasoactive peptides that has been implicated in the progression from edematous to necrotizing pancreatitis is bradykinin. We have investigated the effect of bradykinin administration and bradykinin inhibition on an edematous model of acute pancreatitis in rats (10 micrograms/kg/h of caerulein i.v.). Within six hours i.v. bradykinin reduced circulating serum amylase levels significantly but neither affected tissue edema nor morphology. A bradykinin antagonist (HOE-140), on the other hand, reduced pancreatic edema by 70% and converted edematous pancreatitis into a hemorrhagic and necrotizing variety of the disease. In further experiments we determined the time course and the minimal dosage required for the induction of this severe and non-invasive disease variety. A single dose of caerulein (40 micrograms/kg i.p.) together with a single administration of the bradykinin antagonist HOE-140 (100 micrograms/kg s.c.) consistently resulted in hemorrhagic necrosis of the pancreas within six hours. We conclude that this simple protocol allows for the non-invasive induction of a vascular model of necrotizing pancreatitis and appears ideally suited to study the development of this severe form of the disease.

Topics: Acute Disease; Animals; Anti-Inflammatory Agents, Non-Steroidal; Bradykinin; Ceruletide; Disease Models, Animal; Edema; Hemorrhage; Male; Necrosis; Pancreatitis; Rats; Rats, Wistar

It has been hypothesized that microcirculatory disturbance plays an important role in the development of severe pancreatitis. In this study we investigated the effects of exogenous endothelin-1 on the development of severe pancreatitis in rats.. Acute pancreatitis was induced by two intraperitoneal injections of cerulein (10 micrograms/kg body weight). Endothelin-1 was administered via an abdominal aortic catheter as a bolus of 250-750 pmol/kg BW every hour for 4 h.. Remarkable morphologic changes in the pancreas, including hemorrhage, and increases in serum amylase level and active elastase content in pancreatic tissue were observed in rats treated with cerulein plus endothelin-1 in a dose-dependent manner 5 h after the first cerulein injection. Local pancreatic blood flow decreased significantly, and microcirculatory disturbances in the pancreas were demonstrated.. These results suggest that endothelin-1 causes pancreatic microcirculatory disturbance and might be a contributing factor in the aggravation of acute pancreatitis.

Topics: Acute Disease; Amylases; Animals; Ceruletide; Endothelins; Hemorrhage; Male; Microcirculation; Pancreas; Pancreatic Elastase; Pancreatitis; Rats; Rats, Sprague-Dawley; Regional Blood Flow

The protective effects of a neutrophil elastase inhibitor (ONO-5046) on cerulein-induced pancreatitis followed by a septic challenge with intraperitoneal lipopolysaccharide (LPS) were studied in a rat model. Pancreatitis was induced by four intramuscular injections of cerulein (50 micrograms/kg at 1-hr intervals). ONO-5046 was administered by continuous intravenous infusion via the right jugular vein (50 mg/kg/hr, 30 min prior to the first cerulein injection to 20 hr following the last cerulein injection). Significant differences in serum amylase and pancreatic wet weight ratio were not observed between the animals with pancreatitis treated with or without ONO-5046. There was no significant difference in the in vitro tumor necrosis factor-alpha (TNF-alpha) production by peritoneal macrophages from rats with pancreatitis treated with or without ONO-5046. In a second experiment, LPS (10 mg/kg) was administered intraperitoneally as the septic challenge 6 hr following the first cerulein injection. Lung hemorrhage was seen in the animals with pancreatitis untreated with ONO-5046 24 hr following the first cerulein injection. No significant lung hemorrhage was observed in the animals with pancreatitis treated with ONO-5046 administering 30 min prior to the first cerulein injection. These results suggest that lung hemorrhage in cerulein-induced pancreatitis that follows a septic challenge with LPS can be prevented by the intravenous administration of ONO-5046. Thus there is a significant role for neutrophil elastase in pancreatitis-associated lung injury.

Topics: Animals; Ceruletide; Disease Models, Animal; Drug Evaluation, Preclinical; Glycine; Hemorrhage; Leukocyte Elastase; Lipopolysaccharides; Lung Diseases; Macrophages, Peritoneal; Male; Organ Size; Pancreas; Pancreatic Elastase; Pancreatitis; Rats; Rats, Wistar; Serine Proteinase Inhibitors; Sulfonamides; Time Factors; Tumor Necrosis Factor-alpha

Topics: Acute Disease; Animals; Blood Flow Velocity; Ceruletide; Disease Models, Animal; Endothelin Receptor Antagonists; Endothelins; Hemorrhage; Male; Pancreatitis; Peptides, Cyclic; Rats; Rats, Sprague-Dawley; Stress, Physiological

The role of oxygen-derived free radicals in the pathogenesis of acute pancreatitis was studied in a new model of acute hemorrhagic pancreatitis and cerulein-induced edematous pancreatitis in rats. Hemorrhagic pancreatitis was produced by administering two intraperitoneal doses of cerulein [40 micrograms/kg body weight (BW)] at 1-h intervals following water immersion stress applied for 5 h. Edematous pancreatitis was induced by injecting cerulein as described but without water immersion. Five hours after the first injection of cerulein, pancreatic edema and elevation of serum amylase level were more marked in the animals with hemorrhagic than with edematous pancreatitis. Five hours after the first injection of cerulein, marked hemorrhage and venous dilatation were observed only in those with hemorrhagic pancreatitis. Local pancreatic blood flow decreased to approximately 60% of control values in the animals with edematous pancreatitis, and to approximately 30% of control values in those with hemorrhagic pancreatitis. To evaluate the involvement of oxygen radicals, some rats received three intraperitoneal injections of superoxide dismutase (SOD 10,700 U/kg BW) and catalase (132,000 U/kg BW) beginning 15 min before the first injection of cerulein and repeated at 1-h intervals. No significant effect of free radical scavengers was observed on the edematous pancreatitis. However, in hemorrhagic pancreatitis, treatment with SOD and catalase completely suppressed the hemorrhage and venous dilatation of the pancreas, significantly reduced the pancreatic wet weight and the serum amylase level, and reduced the histologic alterations. However, after treatment with SOD and catalase, no differences were observed in local pancreatic blood flow.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Amylases; Animals; Catalase; Ceruletide; Edema; Free Radicals; Hemorrhage; Immersion; Male; Organ Size; Oxygen; Pancreas; Pancreatitis; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Stress, Physiological; Superoxide Dismutase

We studied morphologic changes in a rat model of acute hemorrhagic pancreatitis in order to investigate the mechanism by which water immersion stress injures the pancreas. Acute hemorrhagic pancreatitis was induced by two intraperitoneal injections of 40 micrograms/kg body weight of caerulein at 1-h intervals under water immersion stress for 5 h at 23 degrees C. Light microscopy showed interstitial edema with inflammatory cell infiltration, degeneration and necrosis of acinar cells, and bleeding. Electron microscopy showed large autophagic vacuoles, decreased zymogen granules, and dilated rough endoplasmic reticulum in acinar cells. Basolateral exocytosis of large vacuoles and phagocytosis of the degenerated acinar cells were observed. In addition, microvascular damage, including the destruction of the capillary endothelial cells, capillary thrombosis, and the extravasation of blood cells, was seen. In contrast, in a pancreatitis model induced by caerulein injection alone, there was no bleeding, no remarkable vascular change, and no thrombosis. Degeneration and necrosis of acinar cells were less severe. In the pancreas under stress alone, microvascular damage and degeneration of acinar cells were observed. These findings demonstrate that stress injures the pancreas and worsens the pancreatitis by causing microcirculatory disturbances, such as vascular damage, thrombosis, increased vascular permeability, and bleeding. These results suggest that chemical mediators, such as free radicals and platelet-activating factor (PAF), which are produced by vascular damage and thrombosis, may accelerate the activation of zymogen proteases in acinar cells in caerulein-induced pancreatitis, leading to hemorrhagic pancreatitis.

Topics: Acute Disease; Animals; Ceruletide; Hemorrhage; Immersion; Male; Microscopy, Electron; Pancreas; Pancreatitis; Rats; Rats, Sprague-Dawley; Stress, Physiological

Topics: Acute Disease; Animals; Ceruletide; Edema; Hemorrhage; Pancreas; Pancreatitis; Rats; Stress, Physiological

The purpose of this study was to investigate whether stress plays a role, morphologically and enzymatically, in the development of severe pancreatitis in rats. Acute hemorrhagic pancreatitis was induced by two intraperitoneal injections of cerulein (40 micrograms/kg body wt) at intervals of 1 h under water-immersion stress for 5 h, whereas water-immersion stress alone did not induce any morphologic and enzymatic changes in the pancreas. In this model, hemorrhagic pancreatitis developed continuously, and the serum amylase level and activation of zymogen proteases in pancreatic tissue were significantly higher than in cerulein-induced pancreatic tissue 5 h after the first cerulein injection. Furthermore, the effects of cerulein on the serum amylase level and activation of zymogen proteases were dose related. Even 5 micrograms/kg body wt of cerulein, which did not induce any evident edematous change in the pancreas, could activate the zymogen proteases of pancreatic tissue fairly well under water-immersion stress compared with pancreatitis induced by 40 micrograms/kg body wt of cerulein alone. These results indicate that stress accelerates the activation of zymogen proteases induced by cerulein and suggest the possibility that stress may play some role in the development of severe pancreatitis.

Topics: Amylases; Animals; Ceruletide; Dose-Response Relationship, Drug; Edema; Enzyme Precursors; Hemorrhage; Immersion; Male; Necrosis; Pancreatic Elastase; Pancreatitis; Rats; Rats, Inbred Strains; Single-Blind Method; Stress, Physiological; Trypsin; Water

Topics: Acute Disease; Animals; Ceruletide; Edema; Hemorrhage; Magnetic Resonance Spectroscopy; Pancreatitis; Rats; Rats, Inbred Strains; Taurocholic Acid

The appearance of vacuoles inside acinar cells characterizes an early stage of development in different models of acute pancreatitis and, possibly, also in human disease. The vacuoles have been shown to contain both digestive and lysosomal enzymes. This abnormal admixture may have important implications for the pathogenesis of pancreatitis because the lysosomal enzyme cathepsin B can activate trypsinogen and may, by this way, trigger pancreatic autodigestion. For the activation process of trypsinogen by cathepsin B, however, an acidic pH is required. This study, therefore, looked for evidence of vacuole acidification in two different models of acute pancreatitis. Edematous pancreatitis was induced in rats by hyperstimulation with cerulein and hemorrhagic pancreatitis was induced in mice by feeding a choline-deficient, ethionine-supplemented diet. Pancreatic acinar cells were isolated at different times after induction of pancreatitis and incubated with 50 microM of acridine orange to identify acidic intracellular compartments. As shown in previous work, zymogen granules are the main acidic compartment of normal acinar cells; they remained acidic throughout the course of pancreatitis in both models. Vacuoles became increasingly more frequent in both models as pancreatitis progressed. Throughout development of pancreatitis, vacuoles accumulated acridine orange indicating an acidic interior. Addition of a protonophore (10 microM monensin or 5 microM carbonyl cyanide m-chlorophenylhydrazone [CCCP] or a weak base (5 mM NH4Cl) completely and rapidly abolished acridine orange fluorescence inside both zymogen granules and vacuoles providing further evidence for an acidic interior. The acidification of vacuoles seen in two different models of pancreatitis may be an important requirement for activation of trypsinogen by cathepsin B and thus for the development of acute pancreatitis.

Topics: Acute Disease; Animals; Cell Compartmentation; Ceruletide; Diet; Female; Hemorrhage; Hydrogen-Ion Concentration; Male; Mice; Organoids; Pancreas; Pancreatitis; Rats; Rats, Inbred Strains; Regression Analysis; Vacuoles

This long-term follow-up of 27 patients treated with conservative surgery for necrohemorrhagic pancreatitis (NHP) showed that an almost complete recovery of the exocrine function is achieved within 4 years after discharge, while about half of the patients presented still abnormal endocrine function. The morphological sequelae, pointed out by endoscopic retrograde pancreatography in almost 50% of the cases, remained unchanged during the follow-up period. Therefore, these data seem to exclude an evolution of NHP towards chronic pancreatitis.

Topics: Acute Disease; Alcoholism; Ceruletide; Female; Follow-Up Studies; Gallstones; Glucose Tolerance Test; Hemorrhage; Humans; Male; Necrosis; Pancreas; Pancreatic Function Tests; Pancreatitis; Postoperative Period; Radiography; Secretin