ceruletide and Pancreatic-Diseases

Although techniques for high-resolution imaging of the pancreas are constantly being improved, the evaluation of pancreatic function remains crucial for the workup of pancreatic diseases. More than 20 direct and indirect tests are available for the assessment of pancreatic function. Measurement of fecal elastase-1 is recommended as the most suitable test for the initial assessment of pancreatic function. Among other techniques, the pancreolauryl test, and alternatively the BT-PABA (N-benzoyl-L-tyrosyl-p-aminobenzoic acid) or the (13)C-mixed-triglyceride test, yield the best sensitivity and specificity. Nevertheless, all indirect tests are of limited value in patients with mild to moderate impairment of pancreatic function. In these patients, the secretin-caerulein test remains the gold standard.

Topics: Ceruletide; Cholangiopancreatography, Endoscopic Retrograde; Humans; Pancreatic Diseases; Pancreatic Function Tests; Secretin

Topics: 4-Aminobenzoic Acid; Amino Acids; Amylases; Ceruletide; Cholecystokinin; Creatine; Feces; Humans; Isoenzymes; Lipase; Lipids; Malabsorption Syndromes; Nitrogen; Pancreas; Pancreatic Diseases; Pancreatic Function Tests; Pancreatic Juice; Radionuclide Imaging; Secretin

Topics: Biliary Tract Diseases; Ceruletide; Cholangiography; Cholelithiasis; Duodenal Ulcer; Endoscopy; Gastrins; Gastrointestinal Diseases; Gastrointestinal Hemorrhage; Gastrointestinal Hormones; Humans; Intestinal Absorption; Lactose Intolerance; Pancreatic Diseases; Peptic Ulcer; Radionuclide Imaging; Secretin; Stomach Neoplasms

Chronic pancreatitis (CP) is a pancreatic inflammatory disease associated with histological changes, including fibrosis, acinar cell loss and immune cell infiltration, and leads to damage of the pancreas, which results in pain, weight loss and loss of pancreas function. Catechin or catechin hydrate (CH) has antioxidant, anticancer and immune‑regulatory effects. However, unlike other catechins, the antifibrotic effects of (+)‑CH have not been widely studied in many diseases, including CP. Therefore, the anti‑fibrotic effects of (+)‑CH against CP were evaluated in the present study. To assess the prophylactic effects of CH, (+)‑CH (1, 5 or 10 mg/kg) or ethanol was administered 1 h before first cerulein (50 µg/kg) injection. To assess the therapeutic effects, (+)‑CH (5 mg/kg) or ethanol was administered after cerulein injection for one or two weeks. In both methods, cerulein was injected intraperitoneally into mice once every hour, six times a day, four times a week, for a total of three weeks, to induce CP. The data showed that (+)‑CH markedly inhibited glandular destruction and inflammation during CP. Moreover, (+)‑CH prevented pancreatic stellate cell (PSC) activation and the production of extracellular matrix components, such as fibronectin 1 and collagens, which suggested that it may act as a novel therapeutic agent. Furthermore, the mechanism and effectiveness of (+)‑CH on pancreatic fibrosis were investigated in isolated PSCs. (+)‑CH suppressed the activation of Smad2 and fibrosis factors that act through transforming growth factor‑β (TGF‑β) or platelet‑derived growth factor. These findings suggest that (+)‑CH exhibits antifibrotic effects in cerulein‑induced CP by inactivating TGF‑β/Smad2 signaling.

Topics: Animals; Catechin; Ceruletide; Ethanol; Mice; Pancreas; Pancreatic Diseases; Pancreatitis, Chronic

The aim of this study was to investigate the role and mechanism of miR-155 in regulating autophagy in a caerulein-induced acute pancreatitis (AP) cellular model. GFP-LC3 immunofluorescence assay was performed to detect autophagy vesicle formation in pancreatic acinar cell line AR42J. AR42J cells were transfected with miR-155 mimic, inhibitor, and corresponding controls to explore the effect of miR-155 on autophagy. The protein levels of LC3-I, LC3-II, Beclin-1, and p62 were analyzed by western blot analysis. Dual-luciferase reporter assay was performed to verify the interaction between miR-155 and Rictor (RPTOR independent companion of MTOR complex 2). The results showed that caerulein treatment induced impaired autophagy as evidenced by an increase in the accumulation of p62 together with LC3-II in AR42J cells, accompanied by miR-155 upregulation. Furthermore, miR-155 overexpression aggravated, whereas miR-155 silencing reduced the caerulein-induced impairment of autophagy. Mechanistically, Rictor was confirmed to be a direct target of miR-155, which could rescue the miR-155 overexpression-mediated aggravation of impaired autophagy. Collectively, these findings indicate that miR-155 aggravates impaired autophagy in caerulein-treated pancreatic acinar cells by targeting Rictor.

Topics: Acinar Cells; Autophagy; Cell Line; Ceruletide; Humans; MicroRNAs; Pancreatic Diseases; Rapamycin-Insensitive Companion of mTOR Protein; Transfection

In a continuation of previous work, Reg3γ protein was further evaluated as a biomarker of pancreatic injury using immunohistochemistry in an additional species.. Mice and rats were treated with intraperitoneal cerulein injections, creating acute pancreatic injury. Mice received 2, 4, or 6 doses, and rats received 1, 2, or 3 doses of cerulein creating low, medium, and high treatment groups. Control animals were dosed with phosphate-buffered saline at corresponding volumes and intervals. Groups of 6 animals were killed 1, 3, 6, 24, and 48 hours after final treatments. Reg3γ immunohistochemical staining and image analysis were performed on pancreatic tissue obtained 6, 24, or 48 hours after control or cerulein treatment. Staining was quantified using image analysis software to calculate area of positivity as a percentage of total tissue area.. Percent positivity of Reg3γ in both species rose by 6 hours, peaked by 24 hours across all 3 cerulein doses, and dropped significantly by 48 hours. In high-dose rats with accompanying gene expression data, Reg3γ gene expression corresponded temporally with quantitative staining data.. Reg3γ staining quantified through image analysis showed a time- and dose-response in cerulein-treated mice and rats.

Topics: Abdominal Injuries; Acute Disease; Animals; Ceruletide; Disease Models, Animal; Gene Expression Regulation; Immunohistochemistry; Injections, Intraperitoneal; Male; Mice, Inbred C57BL; Pancreatic Diseases; Pancreatitis-Associated Proteins; Rats, Sprague-Dawley; Time Factors

Bone morphogenetic proteins (BMPs) have an anti-fibrogenic function in the kidney, lung, and liver. However, their role in chronic pancreatitis (CP) is unknown. The aim of this study was to define the anti-fibrogenic role of BMP signaling in the pancreas in vivo under CP induction. Mice with a deletion of BMP type II receptor (BMPR2(+/-)) were used in this study in comparison with wild-type mice. CP was induced by repetitive cerulein injection intraperitoneally for 4 weeks, and the severity of CP was evaluated. Pancreatic stellate cells (PSCs) were isolated from the mice and treated with BMP2 and TGF-β in vitro, and extracellular matrix protein (ECM) production was measured. Smad and mitogen-activated protein kinase (MAPK) signaling was also evaluated. BMPR2(+/-) mice revealed a greater pancreatic fibrosis, PSC activation and leukocyte infiltration after CP induction compared to wild-type mice (P<0.05). Under CP induction, phospho (p)Smad1/5/8 was elevated in wild-type mice and this effect was abolished in BMPR2(+/-) mice; pSmad2 and pp38(MAPK) were further enhanced in BMPR2(+/-) mice compared to wild-type mice (P<0.05). In vitro, BMP2 inhibited TGF-β-induced ECM protein fibronectin production in wild-type PSCs; this effect was abolished in BMPR2(+/-) PSCs (P<0.05). In BMPR2(+/-) PSCs, pSmad1/5/8 level was barely detectable upon BMP2 stimulation, while pSmad2 level was further enhanced by TGF-β stimulation, compared to wild-type PSCs (P<0.05). BMPR2/Smad1/5/8 signaling plays a protective role against cerulein-induced pancreatic fibrosis by inhibiting Smad2 and p38(MAPK) signaling pathways.

Topics: Animals; Bone Morphogenetic Protein Receptors, Type II; Bone Morphogenetic Proteins; Ceruletide; Fibrosis; Mice; Mice, Knockout; Pancreas; Pancreatic Diseases; Severity of Illness Index; Signal Transduction

Bone marrow (BM) cells may transdifferentiate into circulating fibrocytes and myofibroblasts in organ fibrosis. In this study, we investigated the contribution and functional roles of BM-derived cells in murine cerulein-induced pancreatic fibrosis. C57/BL6 female mice wild-type (WT) or Col 1α1(r/r) male BM transplant, received supraphysiological doses of cerulein to induce pancreatic fibrosis. The CD45(+)Col 1(+) fibrocytes isolated from peripheral blood (PB) and pancreatic tissue were examined by in situ hybridization for Y chromosome detection. The number of BM-derived myofibroblasts, the degree of Sirius red staining and the levels of Col 1α1 mRNA were quantified. The Y chromosome was detected in the nuclei of PB CD45(+)Col 1(+) fibrocytes, confirming that circulating fibrocytes can be derived from BM. Co-expression of α-smooth muscle actin illustrated that fibrocytes can differentiate into myofibroblasts. The number of BM-derived myofibroblasts, degree of collagen deposition and pro-collagen I mRNA expression were higher in the mice that received Col 1α1(r/r) BM, (cells that produce mutated, collagenase-resistant collagen) compared to WT BM, indicating that the genotype of BM cells can alter the degree of pancreatic fibrosis. Our data indicate that CD45(+)Col 1(+) fibrocytes in the PB can be BM-derived, functionally contributing to cerulein-induced pancreatic fibrosis in mice by differentiating into myofibroblasts.

Topics: Animals; Bone Marrow Cells; Bone Marrow Transplantation; Cell Movement; Cell Transdifferentiation; Ceruletide; Collagen; Disease Models, Animal; Female; Fibroblasts; Fibrosis; Leukocyte Common Antigens; Male; Mice; Mice, Inbred C57BL; Pancreatic Diseases

The Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway is widely involved in cell migration, apoptosis and inflammation. However, its exact mechanisms in severe acute pancreatitis (SAP) remain unclear. The aim of this study was to explore the activity of the JAK/STAT signaling pathway in pancreatic injury, investigate the functional mechanisms of SAP in vitro, and thus elucidate the underlying therapeutic effects for SAP in vivo. The activation of the JAK-1/STAT-1 signaling pathway and the expessions of TNF-α, IL-1β and IL-6 proteins were investigated in AR42J cells induced with cerulein and treated with either PBS, RPM, or AG490. One group of cells was left untreated as a control group. Subsequently the activity of NF-κB was evaluated. Rats were given RPM or AG490 just before the induction of SAP, the severity of which was assessed at 24 h. The findings revealed that the up-regulated expressions of JAK-1/STAT-1, STAT-3 protein were closely correlated with the transcription of TNF-α, IL-1β, and IL-6 in cerulein-stimulated cells. Administration of RPM or AG490 decreased the activity of NF-κB and inhibited the release of TNF-α, IL-1β, and IL-6. The reflective markers of severity of SAP were also decreased by RPM or AG490 treatment compared to SAP rats. This study indicates that the JAK-1/STAT-1 signaling pathway activity is an early event in pancreatic inflammatory injury. Therefore, early treatment with its inhibitors might be beneficial for attenuation of pancreatic injury in SAP.

Topics: Animals; Ceruletide; Chemokine CXCL9; Gene Expression Profiling; I-kappa B Kinase; Interferon Regulatory Factor-1; Interleukin-1beta; Interleukin-6; Janus Kinase 1; Janus Kinase 2; Male; NF-kappa B; Pancreatic Diseases; Rats; Rats, Sprague-Dawley; Signal Transduction; Sirolimus; STAT Transcription Factors; STAT1 Transcription Factor; Tumor Necrosis Factor-alpha; Tyrphostins; Up-Regulation

Transforming growth factors betas (TGF-betas) are implicated in pancreatic tissue repair but their role in acute pancreatitis is not known. To determine whether endogenous TGF-betas modulate the course of caerulein induced acute pancreatitis, caerulein was administered to wild-type (FVB-/-) and transgenic mice that are heterozygous (FVB+/-) for expression of a dominant negative type II TGF-beta receptor.. After 7 hourly supramaximal injections of caerulein, the pancreas was evaluated histologically and serum was assayed for amylase and lipase levels. Next, the effects of caerulein on amylase secretion were determined in mouse pancreatic acini, and cholecystokinin (CCK) receptor expression was assessed.. The normal mouse pancreas was devoid of inflammatory cells whereas the pancreas from transgenic mice contained lymphocytic infiltrates. Caerulein injection in wild-type mice resulted in 6- and 36-fold increases in serum amylase and lipase levels, respectively, increased serum trypsinogen activation peptide (TAP) levels, gross oedema and a marked inflammatory response in the pancreas that consisted mainly of neutrophils and macrophages. By contrast, FVB+/- mice exhibited minimal alterations in response to caerulein with attenuated neutrophil-macrophage infiltrates. Moreover, acini from FVB+/- mice did not exhibit restricted stimulation at high caerulein concentrations, even though CCK receptor mRNA levels were not decreased.. Our findings indicate that a functional TGF-beta signalling pathway may be required for caerulein to induce acute pancreatitis and for the CCK receptor to induce acinar cell damage at high ligand concentrations. Our results also support the concept that restricted stimulation at high caerulein concentrations contributes to the ability of caerulein to induce acute pancreatitis.

Topics: Acute Disease; Amylases; Animals; Blotting, Northern; Ceruletide; Edema; Ligands; Lipase; Mice; Mice, Transgenic; Oligopeptides; Pancreatic Diseases; Pancreatitis; Receptors, Cholecystokinin; Severity of Illness Index; Signal Transduction; Transforming Growth Factor beta

The renin-angiotensin system (RAS) plays important roles in various pathophysiological processes. However, the role of the RAS in pancreatic fibrosis has not been established. We investigated the role of angiotensin II (ANG II)-ANG II type 1 (AT(1)) receptor pathway in the development of pancreatic fibrosis with AT(1a) receptor-deficient [AT(1a)(-/-)] mice. To induce pancreatic fibrosis, AT(1a)(-/-) and wild-type (WT) mice were submitted to three episodes of acute pancreatitis induced by six intraperitoneal injections of 50 microg/kg body wt cerulein at hourly intervals, per week, for four consecutive weeks. Pancreatic fibrosis was assessed by histology and hydroxyproline content. Pancreatic stellate cell (PSC) activation and the localization of AT(1) receptors were assessed by Western blot analysis for alpha-smooth muscle actin and immunostaining. Transforming growth factor-beta(1) (TGF-beta(1)) mRNA expression in the pancreas was assessed by RT-PCR. Six intraperitoneal injections of cerulein induced acute pancreatitis in both AT(1a)(-/-) and WT mice. There were no significant differences between two groups with regard to serum amylase and histological changes. Pancreatic fibrosis induced by repeated episodes of acute pancreatitis was significantly attenuated in AT(1a)(-/-) mice compared with that in WT mice. This finding was accompanied by a reduction of activated PSCs. Dual-immunofluorescence staining in WT mice revealed that activated PSCs express AT(1) receptors. The level of TGF-beta(1) mRNA was lower in AT(1a)(-/-) mice than in WT mice. Our results demonstrate that the ANG II-AT(1) receptor pathway is not essential for the local pancreatic injury in acute pancreatitis but plays an important role in the development of pancreatic fibrosis through PSC activation and proliferation.

Topics: Animals; Ceruletide; Fibrosis; Hydroxyproline; Mice; Mice, Inbred C57BL; Mice, Knockout; Pancreas; Pancreatic Diseases; Protein Isoforms; Receptor, Angiotensin, Type 1; Recurrence; RNA, Messenger; Transforming Growth Factor beta; Transforming Growth Factor beta1

Among the three major cytofilament proteins, keratin (K8/K18/K19) expression increases nearly threefold upon pancreas or liver injury, while actin and tubulin expressions are considered relatively stable. K8/K18 serves essential hepatocyte cytoprotective functions yet appears dispensable in K8-null mouse pancreata, which led us to hypothesize that actin or tubulin expressions may increase after pancreatic injury. Balb/c and FVB/n mice manifested different susceptibility to injury in two pancreatitis models, with significant induction of actin protein (threefold) and RNA after moderate or severe but not mild injury. Alterations in tubulin expression were less prominent. Basally, K8-null and wild-type pancreata expressed similar actin and tubulin levels, while the injury-induced actin protein but not RNA was more pronounced in K8-null mice. K7/K18/K19/K20 were also induced in K8-null mice after injury. Ex vivo, caerulein-triggered pancreatitis caused protein degradation (actin approximately or = tubulin > keratins) and mRNA up-regulation that was blocked by actinomycin-D (act-D) (actin approximately or = tubulin approximately or = keratin) or by NF-kappaB inhibition (keratins > actin approximately or = tubulin). Hence, actin is not as static as previously held and is overexpressed after moderate to severe pancreatic injury while keratins are induced after minimal injury. Keratin and actin induction may serve protective roles in pancreatic injury.

Topics: Actins; Animals; Ceruletide; Cytoskeleton; Dactinomycin; Disease Susceptibility; Female; Immunoblotting; Keratin-8; Keratins; Male; Mice; Mice, Inbred BALB C; Mice, Knockout; NF-kappa B; Pancreas; Pancreatic Diseases; Pancreatitis; Protein Synthesis Inhibitors; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Tubulin; Up-Regulation

Transforming growth factor-beta (TGF-beta) is an important cytokine in the fibrogenesis in many organs, including the pancreas. Using an adenoviral vector expressing the entire extracellular domain of type II human TGF-beta receptor (AdTbeta-ExR), we investigated whether inhibition of TGF-beta action is effective against persistent pancreatic fibrosis, and whether it exerts a beneficial effect on the pancreas in the process of chronic injury. To induce chronic pancreatic injury and pancreatic fibrosis, mice were subjected to three episodes of acute pancreatitis induced by six intraperitoneal injections of 50 microg/kg body weight cerulein at hourly intervals, per week for 3 consecutive weeks. Mice were infected once with AdTbeta-ExR, or with a control adenoviral vector expressing bacterial beta-galactosidase (AdLacZ). Pancreatic fibrosis was evaluated by histology and hydroxyproline content. Activation of pancreatic stellate cells (PSCs) was assessed by immunostaining for alpha-smooth muscle actin. Apoptosis and proliferation of acinar cells were assessed by immunostaining of ssDNA and Ki-67, respectively. Three-week cerulein injection induced pancreatic fibrosis and pancreatic atrophy with proliferation of activated PSCs. In AdTbeta-ExR-injected mice, but not AdLacZ-injected mice, pancreatic fibrosis was significantly attenuated. This finding was accompanied by a reduction of activated PSCs. AdTbeta-ExR, but not AdLacZ, significantly increased pancreas weight after chronic pancreatic injury. AdTbeta-ExR did not change the proportion of proliferating acinar cells, whereas it reduced the number of apoptotic acinar cells. Our results demonstrate that inhibition of TGF-beta action not only decreases pancreatic fibrosis but also protects the pancreas against chronic injury by preventing acinar cell apoptosis.

Topics: Animals; Ceruletide; Chronic Disease; Disease Models, Animal; Fibrosis; Male; Mice; Mice, Inbred BALB C; Mice, Transgenic; Pancreatic Diseases; Receptors, Transforming Growth Factor beta; Transforming Growth Factor beta

Pancreatic caerulein-induced activation of c-Jun NH(2)-terminal kinase (JNK) has been reported, and JNK has been proposed as a mediator during induction of hyperstimulated pancreatitis. CEP-1347 has recently been described as a specific JNK inhibitor. We tested whether CEP-1347 inhibits caerulein-induced pancreatic JNK activation in isolated acini and in vivo. CEP-1347 dose dependently inhibited acinar caerulein-induced JNK activation with nearly complete inhibition at 2 microM but had no effect on digestive enzyme release. For in vivo studies, rats were pretreated with CEP-1347 before caerulein hyperstimulation. For assessment of JNK activation and histological alterations, animals were killed 30 min or 2 and 4 h after caerulein hyperstimulation, respectively. Pancreatic wet weight, serum enzyme levels, and pancreatic activity of p38 and extracellular signal-regulated kinase (ERK) were also determined. Caerulein hyperstimulation strongly activated JNK, p38, and ERK. CEP-1347 pretreatment dose dependently reduced caerulein-induced pancreatic JNK activation without p38 or ERK inhibition. JNK inhibition also reduced pancreatic edema formation and reduced histological severity of pancreatitis. Thus we show that CEP-1347 inhibits JNK activation in vivo and ameliorates caerulein-induced pancreatitis.

Topics: Amylases; Animals; Carbazoles; Ceruletide; Dose-Response Relationship, Drug; Edema; Enzyme Activation; Enzyme Inhibitors; In Vitro Techniques; Indoles; JNK Mitogen-Activated Protein Kinases; Male; Mitogen-Activated Protein Kinases; Pancreas; Pancreatic Diseases; Pancreatitis; Rats; Rats, Sprague-Dawley

To determine changes in serum feline trypsin-like immunoreactivity (fTLI) in response to administration of ceruletide to healthy cats.. 11 healthy cats.. Serum fTLI was determined, using a radioimmunoassay, before and 10, 20, 30, 40, and 50 minutes after IM administration of ceruletide (0.3 mg/kg [0.14 mg/lb]).. Mean +/- SD baseline serum fTLI was 23.1 +/- 4.1 mg/L. There was a statistically significant, but clinically unimportant, increase in serum fTLI 10 and 30 minutes after ceruletide administration.. In healthy cats, administration of ceruletide induced a statistically significant, but clinically unimportant, increase in serum fTLI. Whether responses in cats with exocrine pancreatic disorders would be different is unknown, but results suggest that a ceruletide stimulation test would likely not be useful for differentiating between healthy cats and cats with subclinical chronic exocrine pancreatic disorders.

Topics: Animals; Cat Diseases; Cats; Ceruletide; Pancreatic Diseases; Radioimmunoassay; Trypsin

Adenosine has been shown to modulate various pathophysiologic conditions through receptor-mediated mechanisms. However, the role of adenosine in the pathogenesis of acute pancreatitis has not been described. We examined the effect of adenosine-receptor stimulation or inhibition on the pathologic changes of the pancreas.. Rats received intraperitoneal injections of selective agonists of A1, A2a, and A3 adenosine receptors: 2-chloro-N(6)-cyclopentyladenosine (CCPA), CGS-21680 (CGS), or 1-deoxy-1-[6-[[(3-iodophenyl)methyl]amino]-9H-purin-9-yl]-N-methyl-be ta-D-ribofuranuronamide (IB-MECA), respectively. Serum amylase activity and pathologic changes of the pancreas were evaluated. The effects of a specific A1-receptor antagonist (FK-838) on the pathologic findings of cerulein- and taurocholate-induced pancreatitis were also examined.. Administration of a selective A1 agonist induced hyperamylasemia and morphologic changes in the pancreas characterized by interstitial edema and leukocyte infiltration; neither A2a nor A3 agonist produced such changes. Treatment with an A1-receptor antagonist significantly attenuated the outcome induced by A1 agonist stimulation. In addition, the A1-receptor antagonist significantly ameliorated pancreatic edema in both pancreatitis models, although it did not improve the acinar cell damage of the pancreas or the increase of serum amylase.. Activation of the adenosine A1-receptor pathway may have an important role in the pathogenesis of acute pancreatitis.

Topics: Acute Disease; Adenosine; Amylases; Animals; Ceruletide; Edema; Leukocytes; Male; Pancreas; Pancreatic Diseases; Pancreatitis; Phenethylamines; Purinergic P1 Receptor Agonists; Purinergic P1 Receptor Antagonists; Pyrazoles; Pyridines; Rats; Rats, Wistar; Receptor, Adenosine A3; Receptors, Purinergic P1; Taurocholic Acid

The aim of this study was to evaluate the controversial specificity of the plasma amino acid (AA)-consumption test in detecting pancreatic diseases by using two different quantitative methods. A total of 55 subjects: 13 healthy subjects, 13 patients with chronic pancreatitis (three mild/moderate, eight severe), 13 patients with pancreatectomy and complete suppression of the exocrine pancreatic secretion, eight patients with chronic liver disease (five with impaired synthetic function), and eight patients with chronic renal failure. Total plasma AAs were quantified by a colorimetric method (p-benzoquinone) in all subjects, at 0, 30, 45, and 60 min during and 30 min after minute 60 of i.v. cerulein infusion (50 ng/kg/h). Either total and individual AAs were quantified by chromatography (high-performance liquid chromatography; HPLC) in 10 healthy subjects, 10 patients with pancreatectomy, and 10 with chronic pancreatitis at 0 and 60 min after the start of the cerulein infusion. For the colorimetric method, healthy subjects had maximal percentage decreases of total AA concentrations not significantly different from those of patients with pancreatectomy and significantly higher than those of patients with chronic pancreatitis (p < 0.0001) or chronic liver disease (p < 0.001). Pancreatic function, as assessed by fecal elastase-1 test, was not significantly correlated to the maximal percentage decrease in total plasma AAs. For the chromatographic method, total AA concentrations were not significantly correlated to those determined by colorimetry. The concentration of each of the individual plasma AAs varied considerably in each group. Fecal elastase-1 values were normal (> or = 200 microg/g) in all patients without pancreatic disease and in only one of 11 patients with chronic pancreatitis and exocrine insufficiency. The type of method used can explain the different results of the AA-consumption test. This test is not very specific for the pancreas.

Topics: Adult; Aged; Amino Acids; Ceruletide; Chromatography, Ion Exchange; Colorimetry; Evaluation Studies as Topic; Feces; Female; Humans; Kidney Failure, Chronic; Liver Diseases; Male; Middle Aged; Pancreatectomy; Pancreatic Diseases; Pancreatic Elastase; Pancreatic Function Tests; Pancreatitis; Time Factors

Previously we reported that prior administration of lipopolysaccharide (LPS) mitigates subsequently produced cerulein (Cn) pancreatitis. To clarify the mechanism further, the pathological features of Cn pancreatitis were examined in detail after treating rats with very low doses of LPS. LPS pretreatment reduced the formation of pancreatic edema during Cn pancreatitis in a dose- and time-dependent manner. In contrast, the elevation of serum amylase and the histological findings, including acinar cell vacuolization and infiltration of inflammatory cells, were not affected. The lowest dose of LPS, 500 ng/kg, was sufficient to inhibit pancreatic edema formation completely. LPS at a dose of 5 microg/kg was fully effective when it was given from 30 min to 12 h before the induction of pancreatitis. Pretreatment with tumor necrosis factor-alpha (TNF-alpha) inhibited the pancreatic edema in a manner similar to that of LPS. Moreover, the inhibitory effect of LPS was partially attenuated by the administration of anti-TNF-alpha antibody before the injection of LPS. Actinomycin D (0.5 mg/kg) abolished the effect of LPS, whereas cycloheximide (0.5 mg/kg) given alone reduced pancreatic edema formation during pancreatitis. From these results, it was concluded that very low doses of LPS can induce, partially via TNF-alpha, a state refractory to pancreatic edema formation during Cn pancreatitis, and this phenomenon seems to be regulated at the transcriptional level.

Topics: Animals; Ceruletide; Cycloheximide; Dactinomycin; Edema; Lipopolysaccharides; Male; Pancreatic Diseases; Pancreatitis; Rats; Rats, Wistar; Tumor Necrosis Factor-alpha

This study was designed to evaluate the protective effect of a peptide leukotriene receptor antagonist, pranlukast hydrate, against pancreatic injuries during acute pancreatitis.. Acute pancreatitis was induced in rats by intravenous infusion of a supramaximal dose of cerulein (5 micrograms/kg h for 4 h). In this model marked hyperamylasemia, a significant increase in pancreatic water content, and a significant increase in pancreatic microvascular leakage of Evans blue dye were observed. Pancreatic subcellular redistribution of the lysosomal enzyme cathepsin B from the lysosomal fraction to the zymogen fraction was also observed.. Pretreatment with pranlukast hydrate at a dose of 10 micrograms/kg (twice, 8 and 4 h before cerulein infusion) significantly inhibited these pancreatic injuries, including hyperamylasemia, increased pancreatic microvascular permeability, and redistribution of cathepsin B in pancreatic acinar cells.. These results suggest that peptide leukotrienes may be involved in the pathogenesis of acute pancreatitis in the early stage of the disease and that peptide leukotriene receptor antagonist might be of therapeutic value for treatment of acute pancreatitis.

Topics: Acute Disease; Amylases; Animals; Body Water; Cathepsin B; Ceruletide; Chromones; Edema; Leukotriene Antagonists; Male; Pancreas; Pancreatic Diseases; Pancreatitis; Rats; Rats, Wistar

Acute pancreatitis is characterized by hyperamylasemia, pancreatic edema, and the presence of activated digestive enzymes within the pancreas. The secretagogue-induced model of acute pancreatitis is also characterized by pancreatic acinar cell vacuolation, subcellular redistribution of lysosomal hydrolases, and a fall in pancreatic glutathione levels. We have performed time-dependence studies to determine the sequence with which these phenomena appear and to establish their cause-and-effect relationship. Evidence of lysosomal enzyme redistribution and trypsinogen activation within the pancreas could be detected within 10-15 min of the onset of supramaximal secretagogue stimulation, while hyperamylasemia (30 min), pancreatic edema (60 min), and acinar cell vacuolation (60 min) occurred at later times. Pancreatic glutathione levels were either unchanged (15 and 30 min) or elevated (60 min) during the early times of supramaximal stimulation and were only noted to be decreased at a later time. These results support the conclusion that intrapancreatic digestive enzyme activation, possibly occurring by a mechanism involving lysosomal hydrolase redistribution, is an early and likely a critical event in the evolution of secretagogue-induced pancreatitis but that glutathione depletion is neither early nor critical to the evolution of this model of pancreatitis.

Topics: Amylases; Animals; Cathepsin B; Ceruletide; Edema; Enzyme Activation; Glutathione; Male; Pancreas; Pancreatic Diseases; Pancreatitis; Rats; Rats, Wistar; Subcellular Fractions; Trypsinogen

During a secretin-cerulein test for exocrine pancreatic function, venous acid-base analysis was performed before and 30 and 45 min after the intravenous injection of secretin (1 clinical unit/kg body weight). A decrease in plasma bicarbonate was observed in persons with normal exocrine pancreatic function as defined by normal bicarbonate and enzyme secretion in the secretin-cerulein test (n = 39). Bicarbonate values were 94.8 +/- 3.6% (mean +/- SD) of the initial concentration after 30 min and 92.9 +/- 3.9% after 45 min. In contrast, there was no statistically significant decrease in plasma bicarbonate in patients with global pancreatic exocrine deficiency (n = 15). In normal exocrine pancreatic function, the maximal plasma bicarbonate decrease was 2.17 +/- 1.12 mM (mean +/- SD; n = 39). In patients with global exocrine deficiency, however, a maximal plasma bicarbonate decrease of 0.54 +/- 0.68 mM (n = 15) was obtained. The difference was highly significant (p < 0.001). In patients with partial exocrine deficiency (isolated decrease in bicarbonate secretion or reduced excretion of a single enzyme), values were also significantly lower (1.44 +/- 1.40 mM; n = 14; p < 0.05). The maximal plasma bicarbonate decrease correlated with the bicarbonate secretion into the duodenal juice (r = 0.62, p < 0.001). If a maximal plasma bicarbonate decrease > or = 0.9 mM was considered normal, determination of plasma bicarbonate following secretin administration allowed us to detect the presence of global exocrine deficiency with a sensitivity of 87% and a specificity of 87%. The secretin-induced plasma bicarbonate decrease may therefore be used as a new simple tubeless test to evaluate exocrine pancreatic function.

Topics: Acid-Base Equilibrium; Amylases; Bicarbonates; Ceruletide; Chymotrypsin; Female; Humans; Kinetics; Lipase; Male; Pancreas; Pancreatic Diseases; Secretin; Trypsin

In several species, bicarbonate and calcium concentrations of pancreatic juice are known to vary during the different phases of pancreatic secretion. The effects of these variations on the saturation of juice with calcium carbonate, a critical factor for the formation of pancreatic stones, are not known. In this work, we studied the saturation degree of pancreatic juice with calcium carbonate in six unanesthetized beagle dogs equipped with Thomas cannulae during basal secretion and after bolus injections of cerulein (30 ng/kg) or secretin (0.25 units/kg). In the different samples of pure pancreatic juice, pH, PCO2, bicarbonate, and proteins were measured by standard methods. Total calcium (CaT) and ionized calcium (Ca2+) were determined using calcium-specific electrodes. Saturation with calcium carbonate was calculated by reference to the solubility product of calcite at 37 degrees C. Almost all the samples were found to be supersaturated with calcium carbonate but large variations of the saturation index were observed. In basal samples, obtained during periods of low secretion rate, the mean saturation index (3.35 +/- 3.01) was significantly lower than under secretion (12.10 +/- 5.14) or cerulein (18.01 +/- 8.42). This low basal saturation index, in spite of a high Ca2+ content, was explained by a low bicarbonate concentration (37.6 +/- 18.9 mmol/liter) and a high PCO2 (13.4 +/- 7.5 kPa). In contrast, in juice obtained after hormonal stimulation, PCO2 (4.8 +/- 1.6 kPa) was similar to plasma PCO2 (5.5 +/- 1.2 kPa).(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Calcium Carbonate; Calculi; Ceruletide; Dogs; Pancreatic Diseases; Pancreatic Ducts; Pancreatic Juice; Secretin; Time Factors

To find out if and when lysosomal enzymes are excreted into pancreatic juice in physiological and pathological conditions, the changes in the secretion of cathepsin B into pancreatic juice were investigated in 66 Wistar rats with cannulation of common pancreatic-biliary duct and common bile duct, and infusions of caerulein and secretin. In a separate experiment ducts were cannulated and secretin infused as before, but in one group the ducts were "obstructed" and in another they were allowed to remain patent. Obstruction of the pancreatic duct for three hours caused a moderate significant rise in serum amylase activity. Cathepsin B activity in the pancreatic subcellular fractions was redistributed, and the amount of cathepsin B increased. In rats with obstructed ducts the secretion of cathepsin B and other lysosomal enzymes that were stimulated by caerulein was significantly greater than in the animals in which the ducts remained patent. Lysosomal enzymes associated with zymogen granules are secreted into pancreatic juice together with digestive enzymes after stimulation by gut hormones, and they may have pathophysiological roles in pancreatic juice.

Topics: Amylases; Animals; Cathepsin B; Ceruletide; Dose-Response Relationship, Drug; Lysosomes; Pancreatic Diseases; Pancreatic Ducts; Pancreatic Juice; Rats; Rats, Inbred Strains; Secretin; Time Factors

The short-term effects of rat pancreatic duct obstruction were evaluated and compared with those recently reported to follow obstruction of the rabbit pancreatic duct. In both species pancreatic edema and hyperamylasemia are noted, and the lysosomal hydrolase cathepsin B is redistributed from the lysosome-enriched to the zymogen granule-enriched subcellular fraction. Theoretically, this redistribution phenomenon might lead to digestive enzyme activation because cathepsin B is known to be capable of activating trypsinogen. The hyperamylasemia and pancreatic edema (but not the cathepsin B redistribution) that follow rat pancreatic duct obstruction were increased by infusion of a submaximally stimulating dose of the cholecystokinin analogue cerulein. Administration of the cholecystokinin-receptor antagonist L-364,718 reduced the hyperamylasemia but did not alter the pancreatic edema or cathepsin B redistribution. These observations indicate that cholecystokinin may modulate some but not all of the effects of duct obstruction. Secretin administration increased the degree of pancreatic edema and had no demonstrable protective effect. The rat duct-obstruction model described in this report may prove particularly useful in future studies designed to clarify the early events underlying the development of acute pancreatitis.

Topics: Amylases; Animals; Benzodiazepinones; Cathepsin B; Ceruletide; Cholecystokinin; Devazepide; Ligation; Male; Pancreatic Diseases; Pancreatic Ducts; Rats; Rats, Inbred Strains; Secretin; Subcellular Fractions; Time Factors

Lysosomal hydrolases such as cathepsin B are apically secreted from rabbit pancreatic acinar cells via a regulated as opposed to a constitutive pathway. Intravenous infusion of the cholecystokinin analogue caerulein results in highly correlated apical secretion of digestive and lysosomal enzymes, suggesting that they are discharged from the same presecretory compartment (zymogen granules). Lysosomal enzymes appear to enter that compartment as a result of missorting. After 7 h of duct obstruction is relieved, caerulein-stimulated apical secretion of cathepsin B and amylase is increased, but the ratio of cathepsin B to amylase secretion is not different than that following caerulein stimulation of animals never obstructed. These findings indicate that duct obstruction causes an increased amount of both lysosomal and digestive enzymes to accumulate within the secretagogue releasable compartment but that duct obstruction does not increase the degree of lysosomal enzyme missorting into that compartment. Pancreatic duct obstruction causes lysosomal hydrolases to become colocalized with digestive enzymes in organelles that, in size and distribution, resemble zymogen granules but that are not subject to secretion in response to secretagogue stimulation. These organelles may be of importance in the development of pancreatitis.

Topics: Amylases; Animals; Cathepsin B; Ceruletide; Dose-Response Relationship, Drug; Female; Lysosomes; Male; Pancreas; Pancreatic Diseases; Pancreatic Ducts; Rabbits; Secretory Rate

The pathogenesis of acute pancreatitis is incompletely defined, but the outcome is determined in part by an acute inflammatory process. Pancreatitis-associated inflammation appears to play a role in the local retroperitoneal injury as well as in the associated dysfunction of remote organs such as the lung. Tumor necrosis factor (TNF) appears to be a proximal mediator of the inflammatory response. In this study, anti-TNF antibody was administered to rats with caerulein-induced pancreatitis to determine if the observed increases in pancreatic and pulmonary microvascular permeability were related to plasma TNF activity. In contrast to the expected findings, blockade of TNF activity was found to increase the amount of edema formation in both the pulmonary and pancreatic microvascular beds. The mechanism is not known; however, blockade of TNF-induced down regulation of phagocytic cell activity, ablation of TNF-dependent feedback inhibition of other cytokines, failure of induction of endogenous antioxidant systems, or inactivation of the TNF control of microvascular tone are all possible explanations. This is potentially an important observation as clinical strategies are now being developed to modify the inflammatory response in ways presumed advantageous to an injured host.

Topics: Acute Disease; Animals; Antibodies; Ceruletide; Edema; Male; Pancreatic Diseases; Pancreatitis; Pulmonary Edema; Rats; Rats, Inbred Strains; Tumor Necrosis Factor-alpha

This study was performed to assess the effects of misoprostol, a synthetic prostaglandin E1 analog, on cerulein-induced pancreatitis. Per group of 10 each, male Wistar rats received either cerulein (2.5 micrograms/kg/h subcutaneously), cerulein and misoprostol (500 micrograms/kg intraperitoneally at 0 and 4 h), or saline. Rats were killed 6 h after the first injection. Misoprostol treatment significantly reduced interstitial edema and acinar cell lesions induced by hyperstimulation. Pancreatic amylase and chymotrypsin contents were increased by cerulein and returned towards control levels in the misoprostol-treated group. The lysosomal volume density and the pancreatic beta-D-glucuronidase activity were significantly increased after hyperstimulation. The two parameters were significantly reduced by misoprostol. A protective effect of misoprostol against lesions induced by cerulein hyperstimulation would be a consequence of a lysosomal stabilizating effect.

Topics: Acid Phosphatase; Acute Disease; Alprostadil; Amylases; Animals; Ceruletide; Chymotrypsin; Edema; Glucuronidase; Male; Microscopy, Electron; Misoprostol; Organ Size; Pancreatic Diseases; Pancreatitis; Prostaglandins E, Synthetic; Rats; Rats, Inbred Strains

Previous studies have demonstrated that intravenous catalase infusion protects against the formation of pancreatic edema in cerulein-induced acute pancreatitis; however, polyethylene glycol (PEG)-conjugated catalase given as a bolus was not protective. Using radiolabeled catalase and PEG-catalase in subtherapeutic tracer doses, the pancreas tissue distributions of each were determined in rats with and without pancreatitis. Rats with cerulein-induced pancreatitis developed tissue concentrations of catalase within the pancreas that were three times those of PEG-catalase. The relatively low levels of PEG-catalase in the pancreas outside of the vascular compartment suggest that the failure to prevent edema formation may result from inability of PEG-catalase to reach extravascular sites of injury because of the large molecular size.

Topics: Acute Disease; Animals; Catalase; Ceruletide; Drug Combinations; Edema; Male; Pancreas; Pancreatic Diseases; Pancreatitis; Polyethylene Glycols; Rats; Rats, Inbred Strains

We have compared responsiveness of serum lipase and amylase activity to the pancreatic exocrine stimulation with cerulein and secretin (CS test) in normal subjects and patients with pancreas-related and other diseases. The lipase and amylase activities were measured by a sensitive colorimetric method, the BALB-DTNB method and the Caraway method, respectively. The percentage of positive lipase and amylase response cases was as follows: confirmed chronic pancreatitis (N = 22), 27 and 14%; suspected chronic pancreatitis (N = 37), 46 and 32%; pancreatic cancer (N = 16), 44 and 25%; biliary tract diseases (N = 11), 14 and 14%; miscellaneous (N = 11), 0 and 18%; normal subjects (N = 13), and partial pancreatectomy (N = 5), 0 and 0%, respectively. The serum lipase response cannot be regarded as specific for pancreatic diseases because the lipase response cases were found in biliary tract diseases as well. However, in view of frequent, fast, and intense responsiveness to the CS test, the serum lipase activity measured by the BALB-DTNB method may be more useful than the serum amylase as an auxiliary diagnostic aid for suspected pancreatitis which might develop into confirmed chronic pancreatitis or cancer of the head or body of the pancreas.

Topics: Adolescent; Adult; Aged; alpha-Amylases; Biliary Tract Diseases; Ceruletide; Clinical Enzyme Tests; Colorimetry; Female; Humans; Lipase; Male; Middle Aged; Pancreas; Pancreatic Diseases; Pancreatic Function Tests; Pancreatic Neoplasms; Secretin

In a previous study, we found that pancreatic stones could be dissolved in vitro by a citrate solution. We then decided to look for the possible secretion of citrate into the pancreatic juice, and to find a means of increasing it. Pancreatic juice secretion was collected in Thomas fistula dogs submitted to four protocols: basal conditions, secretin stimulation (0.5 and 2.0 U . kg-1 . h-1), caerulein boluses (25 and 100 ng . kg-1) superimposed on 0.5 U . kg-1 . h-1 secretion and intraduodenal citrate infusion (1.05 mmol . kg-1 . h-1). Under basal conditions mean citrate output per 15 min was 0.11 +/- 0.02 mumol. During secretin and caerulein stimulation, citrate output paralleled that of protein, and a linear regression was calculated: Ycitrate = -0.013 + 0.019 Xprotein (mumol/mg). Intraduodenal citrate infusion induced a sharp increase in citrate output which reached 11.5 mumol/15 min, but only 0.12% of the infused citrate amount was secreted into pancreatic juice during 1 h.

Topics: Animals; Calculi; Ceruletide; Citrates; Dogs; Female; Male; Pancreatic Diseases; Pancreatic Juice; Secretin

Plasma cyclic AMP levels were determined during a 40 minute secretin infusion (1 Cl.U kg-1h-1) followed by a 40 minute combined secretin (1 Cl.U kg-1h-1) caerulein (75 ng kg-1h-1) infusion. In nine healthy subjects, both secretin alone and secretin in combination with caerulein did not affect plasma cyclic AMP levels. The same was observed in six patients with chronic pancreatitis. By contrast, in patients suffering from liver disease (nine cases) or extrahepatic cholestasis (six cases), secretin elicited large increases in plasma cyclic AMP concentration; the mean values attained being, respectively, seven and four times higher than before the infusion. On the other hand, increases in plasma cyclic AMP 10 minutes after a bolus injection of glucagon (1 mg) were four times lower in the liver disease group as compared to the controls. The results reported here suggest that the liver plays a major role in the degradation of plasma cyclic AMP produced by target tissues responding to secretin, and in the release of cyclic AMP under glucagon. Liver disease reduce the capacity of the liver to clear cyclic AMP from the blood. The pancreas does not contribute significantly to the cyclic AMP in the blood.

Topics: Ceruletide; Cholestasis; Cyclic AMP; Female; Glucagon; Humans; Liver Diseases; Male; Pancreatic Diseases; Secretin

Topics: Adult; Ceruletide; Female; Humans; Male; Middle Aged; Pancreas; Pancreatic Diseases; Pancreatic Juice; Secretin

Comparative studies between activity of serumlipase, lipaseevocationstest and endoscopic retrograde cholangiopancreaticography (ERCP) in 45 patients with chronic pancreatitis, pancreatic cysts or pancreatic carcinoma confirm, that elevated enzyme values after caerulein stimulation indicate an alteration of the pancreatic parenchyma. However, this findings does not give information on the etiology of the damage. On the basis of the presented results it seems that the lipase-evocationstest has only limited value as screening method within the total of clinical methods for the assessment of light and medium severe stages of chronic pancreatitis and accompanying forms of pancreatitis. There is no evidence that this method may be used for screening for carcinoma of the pancreas. A positive result gives a valuable information, but the lack of an increase of lipase activity does not exclude the existence of pancreatic affection. In each case with clinical suspicion for pancreatic disease further diagnostic investigation by the ERCP and additional radiographic methods should be performed.

Topics: Adult; Aged; Amylases; Ceruletide; Cholangiography; Chronic Disease; Clinical Enzyme Tests; Endoscopy; Humans; Lipase; Male; Middle Aged; Pancreatic Cyst; Pancreatic Diseases; Pancreatic Neoplasms; Pancreatitis

Topics: Ceruletide; Cholecystokinin; Chronic Disease; Endoplasmic Reticulum; Golgi Apparatus; Humans; Insulin; Insulin Secretion; Microscopy, Electron; Pancreas; Pancreatic Diseases; Pancreatic Juice

Topics: Adult; Amylases; Ceruletide; Cholecystokinin; Female; Humans; Male; Middle Aged; Pancreas; Pancreatic Diseases; Secretin