ceruletide and Alcoholism

Alcohol abuse, a main cause of pancreatitis, has been known to augment NF-κB activation and cell necrosis in pancreatitis. However, the underlying mechanisms are unclear. We recently reported that inhibition of protein kinase D (PKD) alleviated NF-κB activation and severity of experimental pancreatitis. Here we investigated whether PKD signaling mediated the modulatory effects of alcohol abuse on pathological responses in alcoholic pancreatitis.. Alcoholic pancreatitis was provoked in two rodent models with pair-feeding control and ethanol-containing Lieber-DeCarli diets for up to 8 weeks followed by up to 7 hourly intraperitoneal injections of cerulein at 1 μg/kg (rats) or 3 μg/kg (mice). Effects of PKD inhibition by PKD inhibitors or genetic deletion of pancreatic PKD isoform (PKD3Δpanc mice) on alcoholic pancreatitis parameters were determined.. Ethanol administration amplified PKD signaling by promoting expression and activation of pancreatic PKD, resulted in augmented/promoted pancreatitis responses. Pharmacological inhibition of PKD or with PKD3Δpanc mice prevented the augmenting/sensitizing effect of ethanol on NF-κB activation and inflammatory responses, cell necrotic death and the severity of disease in alcoholic pancreatitis. PKD inhibition prevented alcohol-enhanced trypsinogen activation, mRNA expression of multiple inflammatory molecules, the receptor-interacting protein kinase activation, ATP depletion, and downregulation of pro-survival Bcl-2 protein in alcoholic pancreatitis. Furthermore, PKD inhibitor CID755673 or CRT0066101, administrated after the induction of pancreatitis in mouse and rat alcoholic pancreatitis models, significantly mitigated the severity of pancreatitis.. PKD mediates effect of alcohol abuse on pathological process of pancreatitis and constitutes a novel therapeutic target to treat this disease.

Topics: Adenosine Triphosphate; Alcoholism; Animals; Ceruletide; Ethanol; Mice; Necrosis; NF-kappa B; Pancreatitis, Alcoholic; Protein Kinase C; Proto-Oncogene Proteins c-bcl-2; Rats; RNA, Messenger; Trypsinogen

Alcoholic chronic pancreatitis (ACP) is characterized by pancreatic necrosis, inflammation, and scarring, the latter of which is due to excessive collagen deposition by activated pancreatic stellate cells (PSC). The aim of this study was to establish a model of ACP in mice, a species that is usually resistant to the toxic effects of alcohol, and to identify the cell type(s) responsible for production of connective tissue growth factor (CTGF), a pro-fibrotic molecule. C57Bl/6 male mice received intraperitoneal ethanol injections for 3 weeks against a background of cerulein-induced acute pancreatitis. Peak blood alcohol levels remained consistently high in ethanol-treated mice as compared with control mice. In mice receiving ethanol plus cerulein, there was increased collagen deposition as compared with other treatment groups as well as increased frequency of alpha-smooth muscle actin and desmin-positive PSC, which also showed significantly enhanced CTGF protein production. Expression of mRNA for collagen alpha1(I), alpha-smooth muscle actin or CTGF were all increased and co-localized exclusively to activated PSC in ACP. Pancreatic expression of mRNA for key profibrotic markers were all increased in ACP. In conclusion, a mouse model of ACP has been developed that mimics key pathophysiological features of the disease in humans and which shows that activated PSC are the principal producers of collagen and CTGF. PSC-derived CTGF is thus a candidate therapeutic target in anti-fibrotic strategies for ACP.

Topics: Alcoholics; Alcoholism; Animals; Ceruletide; Collagen; Connective Tissue Growth Factor; Ethanol; Extracellular Matrix; Fibrosis; Male; Mice; Mice, Inbred C57BL; Pancreas; Pancreatitis; Pancreatitis, Alcoholic; Pancreatitis, Chronic; RNA, Messenger

Alcohol consumption is a risk factor for chronic pancreatitis (CP), but the mechanism in humans remains obscure because prolonged alcohol consumption in most humans and animal models fails to produce alcoholic chronic pancreatitis (ACP). We hypothesize that the process leading to ACP is triggered by a sentinel acute pancreatitis (AP) event; this event causes recruitment of inflammatory cells, which initiates fibrosis driven by the anti-inflammatory response to recurrent AP and/or chronic oxidative stress. The aim was to determine whether chronic alcohol consumption accelerates fibrosis in response to cerulein-induced pancreatitis in the rat. Wistar male rats were pair-fed control (C) or 5% ethanol (E) Lieber-DeCarli liquid diets. Animals were studied without pancreatitis (P0), with cerulein pancreatitis induced once (P1), or with cerulein-induced pancreatitis weekly for 3 weeks (P3). AP markers, inflammation, and fibrosis were measured histologically, by gene expression profiling and protein expression. Macrophage infiltration was reduced in EP0 versus CP0 rats, but the pattern was reversed after AP. Microabscess, severe necrosis, and early calcification were only induced in the EP3 rats. Fibrosis was significantly induced in the EP3 rats versus EP1, CP1, and CP3 by histology, hydroxyproline content, and mRNA expression for collagen alpha1(1) and procollagen alpha2(1). Proinflammatory cytokine mRNAs were up-regulated shortly after induction of AP, while the anti-inflammatory cytokines (interleukin-10 and transforming growth factor-beta) were strongly up-regulated later and in parallel with fibrogenesis, especially in the EP3 rats. Pancreatic fibrosis develops after repeated episodes of AP and is potentiated by alcohol. Expression of fibrosis-associated genes was associated with expression of anti-inflammatory cytokines in alcohol-fed rats.

Topics: Alcohol Drinking; Alcoholism; Animals; Ceruletide; Chronic Disease; Disease Models, Animal; DNA Primers; Fibrosis; Gene Expression Regulation; Male; Pancreatitis; Polymerase Chain Reaction; Rats; Rats, Wistar; RNA, Messenger

In chronic alcohol abusers with no pancreatic disease, secretin was found to induce a paradoxical spasmodic response in the sphincter of Oddi (SO) instead of the relaxation observed in controls. Cerulein, on the contrary, had a normal relaxing effect on the SO.. We previously reported SO dyskinesia in cases of chronic pancreatitis. Here we investigated whether chronic alcohol consumption may have contributed to the genesis of this dyskinesia.. SO and main pancreatic duct pressures were recorded endoscopically with a dual electronic pressure sensor in 27 chronic alcohol abusers and compared with the values obtained in 15 normal controls. These pressures were recorded both in the basal state and after applying hormonal stimulation by injecting either secretin (1 CU/kg) or cerulein (75 ng/kg).. Cerulein relaxed the SO in both the controls and the chronic alcohol abusers, whereas it transiently enhanced the main pancreatic duct (MPD) pressure. Secretin induced a wave of MPD hyperpressure (+15.4 +/- 3.0 mm Hg) in both groups of subjects, but in the alcoholic group, instead of relaxing SO, it significantly enhanced the amplitude of phasic contractions (+32.6 +/- 8.4 mm Hg). The SO basal pressure was also paradoxically enhanced by secretin in the alcoholic patients (28.8 +/- 8.2 vs 10.1 +/- 2.4 mm Hg).

Topics: Alcoholism; Ceruletide; Cholecystokinin; Dyskinesia, Drug-Induced; Female; Humans; Male; Manometry; Middle Aged; Pancreatic Ducts; Reference Values; Secretin; Sphincter of Oddi

Despite the fact that alcoholism is one of the major causes of pancreatitis, the pathogenesis of this disorder remains obscure. Factors such as the pattern of ethanol consumption, diet, and genetic predisposition may be contributing factors. The failure to produce alcoholic pancreatitis in experimental animals suggests that experimental provision of ethanol may only increase the predisposition to pancreatitis. To test this possibility, we developed an assay system using the in vitro model of cerulein-induced pancreatitis. In this system, pancreatic lobules were first exposed to a supraphysiologic concentration (10(-6) M) of the cholecystokinin analogue, cerulein, after which homogenates were incubated for up to 6 h. Activation of trypsinogen and chymotrypsinogen was observed only in cerulein-treated preparations. We then investigated the effects of the duration of ethanol feeding on cerulein-induced changes in rat pancreas. The pancreata from rats fed ethanol for 9-12 months were more susceptible to cerulein-induced activation of chymotrypsinogen compared to the pancreata from pair-fed control animals. This susceptibility also paralleled morphologic changes, such as dilatation of endoplasmic reticulum, only in the ethanol-fed group. In contrast, during the early stages (up to 3 months) of ethanol consumption, there was resistance (p < 0.01) to cerulein-induced changes. These results suggest that long-term ethanol consumption increases susceptibility to pancreatitis and raises the possibility that a similar mechanism may operate in human alcoholics.

Topics: Alcoholism; Animals; Ceruletide; Chymotrypsinogen; Endoplasmic Reticulum; Enzyme Activation; Ethanol; Male; Microscopy, Electron; Pancreas; Pancreatitis; Rats; Rats, Sprague-Dawley; Trypsin; Trypsinogen

In order to reproduce what might occur during the initial phase in some cases of acute alcohol-induced pancreatitis, rabbits were infused with diluted ethanol and low-dose cerulein. The duct permeability was assessed by recovery of fluoresceinated dextran (molecular weight 19,500) in central venous blood following orthograde duct perfusion with this substance in the anesthetized animal. Serum ethanol, lipase, and amylase were measured; pancreatic duct morphology was examined by light microscopy and electron microscopy. ATP and glutathione were measured, as were amylase, trypsinogen/trypsin, cathepsin B, and DNA levels in differential centrifugates. As expected, acinar amylase and trypsinogen showed a significant decrease in the experimental group; cathepsin B activity was similarly diminished. Compared with the control group, the activity of serum amylase and lipase in the experimental group demonstrated a significant increase. However, no differences between saline-infused control animals and the treated group regarding pancreatic duct permeability, continuity of lumen-lining epithelium, ATP and glutathione levels, and the relative subcellular distribution of pancreatic digestive and lysosomal enzymes were observed. Thus, our findings do not support the relevance of some of the most common hypotheses on the pathophysiology of acute pancreatitis in its early stage for at least a certain subgroup of patients with acute alcohol-induced pancreatitis.

Topics: Adenosine Triphosphate; Alcoholism; Animals; Ceruletide; Ethanol; Glutathione; Male; Pancreas; Pancreatic Ducts; Pancreatitis; Rabbits

Seventeen Indian patients from Kerala State and 13 Indian controls were submitted to a dietary inquiry. Indian patients and controls had a low fat intake (40.8 g +/- 12.1 and 34.5 g +/- 11.0 per day, respectively) and a moderately low protein intake (52.8 +/- 9.5 and 47.8 +/- 11.3 g per day); 11 patients and 6 controls did not consume cassava. Pure nonactivated pancreatic juice was collected at endoscopy in 10 Indian patients who presented with tropical calcific diabetes, 12 apparently normal controls from the same area, and 23 apparently normal French controls. The only significant differences between Indian and French controls was a decreased pancreatic protein response to cerulein and an increased calcium concentration in the Indian subjects. The pancreatic juice of Indian patients was characterized by decreased volume, normal bicarbonate concentration, increased protein concentration when the acinar cells were not stimulated, with no response to cerulein, increased calcium concentration, and normal citrate concentration. These changes are very similar to the changes observed in French patients with chronic alcoholic pancreatitis. The lesions of 14 surgical resection pancreatic specimens from South Indian patients presenting with tropical pancreatitis were compared to pancreata from French patients presenting with chronic alcoholic pancreatitis. The only difference was that intraductal plugs, lesions of the duct epithelium, and retention cysts or pseudocysts were less frequent in Indians. These results show that the two nutritional forms of pancreatic lithiasis, alcoholic and tropical, have similar histological lesions and biochemical modifications of pancreatic juice.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adult; Alcoholism; Calcinosis; Calcium; Ceruletide; Chronic Disease; Citrates; Diet; Female; Humans; India; Lipase; Male; Pancreatic Juice; Pancreatitis; Proteins

Gallbladder dynamics, cholecystokinin (CCK), and pancreatic polypeptide (PP) release were studied in 14 patients with chronic pancreatitis (CP) (2 females, 12 males; age range 24-56 years) and 12 control subjects (4 females, 8 males, 21-50 years). On day 1, gallbladder contractility was investigated after ceruletide intravenous infusion (2.5 ng/kg/min for 10 min). On day 2, a mixed standard test meal (1450 kJ) was administered orally. Gallbladder volume was assessed at three time intervals before (-30, -15, 0 min) and at 5, 10, 20, 30, 40, 50, 60, 80, 100 and 120 min after stimulation by means of ultrasonography. CCK and PP plasma levels were determined at each time interval. Exocrine pancreatic function was assessed using the pancreolauryl serum test (PLT). Six patients with CP had severe exocrine pancreatic insufficiency (EPI) (PLT < 1.8 micrograms/ml) with steatorrhea, eight patients had mild-moderate EPI. Fasting gallbladder volume was increased in CP (32.3 +/- 3.1 cm3) as compared to controls (20.5 +/- 1.2 cm3) (P < 0.01). Peak gallbladder contraction (percent of initial volume) in CP ranged from 5 to 55% (controls: 8-46%) following ceruletide and from 17 to 86% (controls: 27-80%) following the test meal (NS). There was no correlation between the degree of EPI according to PLT and peak gallbladder contraction. Gallbladder emptying in CP patients was not different from controls, although the postprandial CCK response was significantly impaired (P < 0.01). Postprandial PP response in CP was correlated with the PLT result (r = 0.78; P < 0.01) but not with gallbladder emptying or refilling time.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adult; Alcoholism; Ceruletide; Cholecystokinin; Chronic Disease; Female; Food; Gallbladder; Gallbladder Emptying; Humans; Male; Middle Aged; Pancreas; Pancreatic Function Tests; Pancreatic Polypeptide; Pancreatitis; Radioimmunoassay

Pure pancreatic juice composition was studied, after secretin and cerulein stimulation in 29 people from Burundi (Central Africa): 17 controls and 12 alcoholic chronic pancreatitis (CP) patients. Results were compared to similar data in France. African controls had a similar pancreatic response to hormones apart from a much lower lipase secretion than French controls. In the early non-calcified stage of African CP water and bicarbonate secretion were markedly diminished while protein and lipase concentrations were enhanced. In the late stage, secretion was exhausted except that of calcium. Nutritional data were obtained under the same conditions in 40 African controls and in 34 CP patients (including all patients tested for secretion). African controls had a very low fat intake (35.2 +/- 2.6 g/day), and patients had a higher protein and fat intake (144.7 +/- 5.9 and 66.2 +/- 4.8 g/day, respectively) than local controls: as in other countries, CP was associated with a diet enriched in alcohol, fat and protein.

Topics: Adult; Alcoholism; Burundi; Ceruletide; Dietary Fats; Dietary Proteins; Female; France; Humans; Male; Middle Aged; Pancreatic Juice; Pancreatitis; Secretin

187 patients were checked up over 4 years by the secretin-ceruletide test. Independently of the test results they were assigned to various disease groups on the basis of clinical assessment. 131 subjects were divided in a pilot investigation into: subjects with a healthy pancreas (n = 55); subjects with chronic pancreatitis (n = 50); subjects whose pancreatic condition could not be classified clearly (n = 26). 8 parameters were compared by univariate and multivariate statistical procedures in order to confirm or rule out the presence of chronic pancreatitis. The discriminatory power of the following parameters in duodenal fluid proved to be sufficiently high, with less than 15% frequency of misclassification: chymotrypsin (activity) and/or; lipase (activity) and/or; amylase (activity); viscosity. Under routine conditions measurement of the activity of two of these enzymes is sufficient. Their contribution to discrimination proved to be approximately equal. The diagnostic sensitivity and specificity of the parameters bicarbonate, lipase (concentration), trypsin (activity) and volume of duodenal fluid are lower. The classification rules derived from the above pilot group were confirmed by a diagnostic study under routine condition in a test group of 38 patients. Limitation to examining only volume and a maximum of 3 parameters which proved best in distinguishing between patients with chronic pancreatitis and healthy subjects, together with the omission of the first-hour samples after a secretin bolus, considerably reduced laboratory workload without altering the discriminatory power of the secretin-ceruletide test.

Topics: Adult; Alcoholism; Amylases; Ceruletide; Chronic Disease; Chymotrypsin; Duodenum; Female; Humans; Intestinal Secretions; Lipase; Male; Middle Aged; Pancreatitis; Reference Values; Secretin; Viscosity

Rats were chronically fed either an ethanol-containing diet (36% of total calories derived from alcohol) or a pair-fed, control diet (no alcohol) for 8 wk, and acute pancreatitis (AP) was subsequently induced by a 3-h i.v. infusion of caerulein (CR) at a dose of 5 micrograms/kg/hr. CR-induced AP in control rats (no alcohol) was characterized by a significant elevation in serum lipase content, pancreatic interstitial edema, infrequent occurrences of karyorrhexis, and the appearance of vacuoles in acinar cells. Chronic feeding of the ethanol diet followed by treatment with CR resulted in increases in serum lipase content, interstitial edema, karyorrhexis, and acinar vacuolization that were significantly greater than that seen in rats fed the control diet and treated with CR. It is concluded that chronic ethanol intake in the rat intensifies AP that is subsequently induced by CR.

Topics: Acute Disease; Alcoholism; Animals; Ceruletide; Disease Models, Animal; Edema; Lipase; Lysosomes; Male; Pancreas; Pancreatitis; Rats; Rats, Inbred Strains; Vacuoles

Thirty patients suffering from chronic alcoholic pancreatitis (18 calcified) were entered into a study of exocrine and endocrine pancreatic function based on two maximal stimulation tests, namely the secretin-cerulein test and the glucagon test with serum assays of C peptide. The glucagon test was also performed in 19 control subjects. In addition, 10 chronic pancreatitis patients and nine controls were subjected to an oral glucose tolerance test (OGTT) with serum insulin determinations. C peptide basal values were decreased only in patients with severe pancreatic exocrine insufficiency (P less than 0.001), while delta C peptide values were also reduced in patients with moderate exocrine insufficiency (P less than 0.001). Lipase output correlated very well with delta C peptide values (P less than 0.001). While serum insulin levels during OGTT and C peptide basal values showed no significant differences between the chronic pancreatitis and control groups, delta C peptide values were significantly reduced in chronic pancreatitis patients (P less than 0.02). Both endocrine and exocrine function are impaired in chronic pancreatitis, as demonstrated by maximal tests, even in early stages of the disease.

Topics: Adult; Alcoholism; C-Peptide; Ceruletide; Chronic Disease; Female; Glucagon; Glucose Tolerance Test; Humans; Insulin; Islets of Langerhans; Male; Middle Aged; Pancreas; Pancreatic Function Tests; Pancreatitis; Secretin

Seventy-two chronic alcoholics, 40 (all males) with chronic pancreatitis and 32 (23 males and nine females) with liver cirrhosis, were submitted to liver biopsy, endoscopic retrograde cholangiopancreatography and secretin-caerulein test in order to assess a possible liver involvement in chronic pancreatitis and viceversa, and to evaluate the existence of any relationship between the diseases of these two organs. Chronic pancreatitis patients were younger than cirrhotic patients and drank more than the cirrhotic females. Twenty-nine of the 40 patients had abnormal liver histology, five had micronodular cirrhosis and were older than the others. No relationship was found between the degree of pancreatic impairment and the type of liver injury. Five liver cirrhosis patients had an endoscopic retrograde cholangiopancreatography picture consistent with chronic pancreatitis; two were females with an alcohol intake lower than the one of the other females. In conclusion the association of chronic pancreatitis and liver cirrhosis was observed in a minority of cases, with the same percentage in the two groups, even if the cirrhotic subjects were older than the pancreatitics. Therefore we can postulate that different factors have roles in the pathogenesis of alcoholic cirrhosis and of chronic alcoholic pancreatitis. The association of the two diseases in two women with a relatively low alcohol intake supports this hypothesis.

Topics: Adult; Alcoholism; Ceruletide; Cholangiopancreatography, Endoscopic Retrograde; Chronic Disease; Female; Humans; Liver Cirrhosis, Alcoholic; Male; Pancreatic Ducts; Pancreatic Function Tests; Pancreatitis; Prospective Studies; Secretin

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

Previous studies have shown dissolution of human pancreatic stones in vitro by citrate solution which bind considerable amounts of ionized calcium. Pancreatic citrate secretion has been demonstrated in canine and human pancreatic juices. This study compares pancreatic citrate secretion of chronic alcohol-fed dogs with controls in response to graded doses of caerulein, in order to evaluate possible differences in factors favouring pancreatic lithogenicity. The dose-response-relation of citrate outputs after graded doses of caerulein revealed significantly reduced maximal secretory capacity of citrate in alcoholic dogs. Protein concentrations in juices from alcoholic dogs were reduced for all doses of caerulein but protein outputs were not different. Bicarbonate concentrations and outputs, as well as volumes, were significantly greater in alcoholic dogs. Linear relation were found between citrate and protein secretion. Chronic alcohol consumption in the dog leads to reduced citrate secretion, which is consistent with recent results in humans suffering from chronic calcifying pancreatitis, who secrete significantly less citrate than healthy subjects. The decrease of calcium-chelating citrate could be an additional factor causing increased calcium levels in the pancreatic juice of chronic alcoholics, a circumstance that might favour protein-plug formation and subsequently pancreatic stone formation.

Topics: Alcoholism; Animals; Ceruletide; Citrates; Dogs; Dose-Response Relationship, Drug; Female; Humans; Male; Pancreas; Proteins; Secretin

In non-alcoholic dogs the exocrine pancreatic response to caerulein, but not to urecholine or secretin alone, was increased by atropine. This indicated a pancreatic inhibition triggered by the caerulein stimulation and blocked by atropine. The present aim was to examine whether atropine had a similar action on the caerulein-stimulated pancreatic secretion in dogs submitted to long-term alcohol feeding. Alcohol-fed dogs showed an increased volume and bicarbonate response to submaximal caerulein but a non-modified protein response as compared with the responses before alcohol adaptation. The elevated water and bicarbonate responses were not further enhanced by atropine, which, in contrast, enhanced the responses of normal dogs to such an extent as to equalize their responses with those of the treated dogs. Atropine, 5 micrograms X kg-1 X h-1, enhanced similar protein responses to submaximal caerulein more in normal than in alcohol-fed dogs, but with an eightfold higher atropine dose no enhance was produced in dogs alcohol-fed for 36 months. This dose of atropine still evoked a small and similar enhancement in the normal and 9-month-treated dogs. The enhancing action of atropine on caerulein-stimulated secretion, present in normal dogs, hence diminishes and finally disappears as a result of chronic alcohol feeding.

Topics: Alcoholism; Animals; Atropine; Bethanechol Compounds; Ceruletide; Dogs; Duodenum; Humans; Pancreas; Secretin; Stomach

Topics: Adult; Alcoholism; Ceruletide; Chronic Disease; Contrast Media; Female; Hepatic Artery; Humans; Male; Mesenteric Arteries; Middle Aged; Pancreatitis; Radiography