trypsinogen and Alcoholism

Pancreatitis is usually inflammation of the pancreas without infection. Our understanding of pancreatitis has been built on autopsy studies, surgical biopsies and surrogate markers of inflammation and fibroses, including abdominal imaging techniques and pancreatic functional studies. However, the discovery that a number of different environmental factors and various genetic abnormalities are seen in patients with similar appearing pancreatitis phenotypes teaches us that end-stage pathology is not the disorder. Understanding complex associations and interactions requires that the components and their interactions be organized, stratified and functionally defined. Systems biology, in the broad sense, provides the approach and tools to define the complex mechanisms driving pathology. As the mathematics behind these pathways and mechanisms are defined and calibrated, the potential pathology of patients with early signs of disease can be predicted, and a number of patient-specific targets for intervention can be defined.

Topics: Alcoholism; Biomarkers; Cystic Fibrosis Transmembrane Conductance Regulator; Genetic Testing; Humans; Meta-Analysis as Topic; Models, Biological; Pancreas; Pancreatitis; Systems Biology; Trypsinogen

Topics: Acute Disease; Alcoholism; Apoptosis; Cathepsin B; Chemokines; Cholelithiasis; Cytokines; Humans; Necrosis; Oxidative Stress; Pancreatitis; Regeneration; Risk Factors; Trypsin; Trypsinogen

Despite the recent development of medical imaging technology, chronic pancreatitis can only be diagnosed when the disease is fully established. This is due to the lack of specific and sensitive markers for this disease. The discovery of mutations in the cationic trypsinogen gene in patients with hereditary pancreatitis and a high incidence of mutations in the cystic fibrosis transmembrane conductance regulator gene in patients with chronic pancreatitis might be important clues to understanding the molecular mechanisms of this disease. The interaction between ethanol and ion channels might be the missing link between alcohol ingestion and chronic pancreatitis.

Topics: Adult; Aged; Alcoholism; Amino Acid Substitution; Animals; Biomarkers; Chloride Channels; Chronic Disease; Cystic Fibrosis Transmembrane Conductance Regulator; Dogs; Ethanol; Female; Genetic Predisposition to Disease; Humans; Male; Middle Aged; Models, Biological; Pancreatitis; Point Mutation; Protein Conformation; Trypsin; Trypsinogen

Topics: Acute Disease; Alcoholism; Calcinosis; Cholecystitis; Chronic Disease; Enzyme Activation; Humans; Kinins; Lactoferrin; Pancreatitis; Protein Biosynthesis; Protein-Energy Malnutrition; Shock; Trypsin; Trypsinogen

Topics: Acute Disease; Alcoholism; alpha 1-Antitrypsin; alpha-Macroglobulins; Cholelithiasis; Critical Care; Enzyme Activation; Humans; Metabolic Diseases; Pancreatitis; Parasympatholytics; Peritoneal Dialysis; Trypsinogen

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

Topics: Alcoholism; Claudins; Haplotypes; Humans; Nuclear Proteins; Pancreatitis, Chronic; Polymorphism, Genetic; Trypsin; Trypsinogen

Topics: Alcoholism; Animals; Haplotypes; Humans; Mice; Pancreatitis, Chronic; Trypsin; Trypsinogen

Alcohol abuse is a major global health problem, but there is still much uncertainty about the mechanisms of action. So far, the effects of ethanol on ion channels in the plasma membrane have received the most attention. We have now investigated actions on intracellular calcium channels in pancreatic acinar cells. Our aim was to discover the mechanism by which alcohol influences calcium homeostasis and thereby understand how alcohol can trigger premature intracellular trypsinogen activation, which is the initiating step for alcohol-induced pancreatitis. We used intact or two-photon permeabilized acinar cells isolated from wild-type mice or mice in which inositol trisphosphate receptors of type 2 or types 2 and 3 were knocked out. In permeabilized pancreatic acinar cells even a relatively low ethanol concentration elicited calcium release from intracellular stores and intracellular trypsinogen activation. The calcium sensor calmodulin (at a normal intracellular concentration) markedly reduced ethanol-induced calcium release and trypsinogen activation in permeabilized cells, effects prevented by the calmodulin inhibitor peptide. A calmodulin activator virtually abolished the modest ethanol effects in intact cells. Both ethanol-elicited calcium liberation and trypsinogen activation were significantly reduced in cells from type 2 inositol trisphosphate receptor knockout mice. More profound reductions were seen in cells from double inositol trisphosphate receptor (types 2 and 3) knockout mice. The inositol trisphosphate receptors, required for normal pancreatic stimulus-secretion coupling, are also responsible for the toxic ethanol action. Calmodulin protects by reducing calcium release sensitivity.

Topics: Alcoholism; Animals; Calcium; Calcium Channels; Calmodulin; Cells, Cultured; Enzyme Activation; Gene Knockout Techniques; Inositol 1,4,5-Trisphosphate Receptors; Mice; Mice, Transgenic; Pancreas; Trypsinogen

Chronic pancreatitis is an inflammatory disease followed by structural alterations--inflammation, fibrosis and acinar atrophy--pain emergence, exocrine and endocrine pancreatic insufficiency, severe alteration of quality of life. The pathogenetic mechanisms characteristic to this disease are not thoroughly known, but the identification of some genetic and autoimmune factors in certain entities has elucidated several pathogenetic links. The etiologic risk factors for chronic pancreatitis may associate each other and may cause different evolutions to the disease. By tracing out the risk factors and their typical working mechanisms, further pathogenetic treatments may occur, taking place precociously and preventing the evolution of the disease towards exocrine and endocrine pancreatic insufficiency.

Topics: Alcoholism; Autoimmune Diseases; Carrier Proteins; Cystic Fibrosis Transmembrane Conductance Regulator; Humans; Mutation; Pancreatitis, Chronic; Risk Factors; Severity of Illness Index; Smoking; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Several genetic factors have been well known to predispose one to chronic pancreatitis (CP). However, little is known about the genetic factors that may provide a protective effect against the disease. Having found a nonsense mutation (c.111C>A; Y37X) and a splicing mutation (IVS2+1G>A) in the cationic trypsinogen gene (protease, serine, 1; PRSS1) in alcoholics without the development of CP, but not in alcoholics with CP and patients with hereditary or idiopathic CP, we propose that while "gain of function" mutations in the PRSS1 gene predispose one to pancreatitis, "loss of function" mutations in the gene may protect one against the disease.

Topics: Adult; Aged; Alcoholism; Chronic Disease; Exons; Female; Genotype; Humans; Male; Middle Aged; Mutation; Pancreatitis; Trypsin; Trypsinogen

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 rats fed control and ethanol-containing Lieber-DeCarli diets for a period of 12 months, the bile did not contain any enterokinase, the pancreatic juice did not contain any plasmin or thrombin, but in animals fed high fat diet with ethanol, trypsinogen and chymotrypsinogen were significantly increased and trypsin inhibitor decreased. In the tissue, free trypsin and cathepsin B were increased. Composite profile of trypsinogen in gel segments obtained from the pancreatic juice and the tissue showed higher peaks of cationic and anionic variants of trypsinogen in animals fed ethanol. There was no evidence of mesotrypsinogen or of enzyme Y in the juice or the tissue. These studies show that serine proteases and cathepsin B may play a major role in the pathobiology of alcoholic pancreatitis.

Topics: Alcoholism; Animals; Bile; Chymotrypsinogen; Dietary Fats; Enzyme Activation; Ethanol; Pancreas; Pancreatic Juice; Pancreatitis; Rats; Trypsinogen

Topics: Alcoholism; Amylases; Animals; Atrophy; Cholecystokinin; Dietary Fats; Male; Pancreas; Pancreatitis; Rats; Rats, Inbred Strains; Receptors, Cholecystokinin; Trypsinogen

We evaluated the behavior of serum cationic trypsinogen (SCT), an enzyme of solely pancreatic origin, in 30 patients with chronic pancreatitis and 25 healthy subjects as a control, after secretin and bombesin stimulation. After both the stimulations, serum cationic trypsinogen is unable to distinguish between the healthy control subjects and the patients with chronic pancreatitis. On the other hand, after secretin, the enzyme is able to separate chronic pancreatitis patients with different levels of exocrine function insufficiency. It does so with a greater statistical significance than that obtained by the rapid injection of bombesin and equal to that of trypsin into the duodenal juice during duodenal intubation. For these reasons, as well as the absence of any side-effects, secretin is preferred to bombesin stimulation in the evaluation of the exocrine pancreatic function in patients with chronic pancreatitis.

Topics: Adult; Alcoholism; Bombesin; Cations; Chronic Disease; Female; Humans; Male; Middle Aged; Pancreatitis; Secretin; Trypsinogen

The present study was done to determine the effect of the interaction of ethanol and an average or high-fat diet on pancreatic acinar cell function. Weight-matched groups of Sprague-Dawley rats were fed regular rat feed or average fat and high-fat liquid diets with or without 5% (w/v) concentration of ethanol for 3 months. Trypsinogen secretion was increased by the high-fat diet but not by ethanol; chymotrypsinogen secretion was decreased by the combination of ethanol and a high-fat diet, while lipase secretion was increased by a high-fat diet. Pancreatic secretory trypsin inhibitor was not significantly altered by ethanol or fat in the diet. Ethanol feeding together with a high-fat content of the diet caused complex and nonparallel changes in the secretion of the pancreatic enzymes. These data are of interest in view of the reported positive correlation between alcoholism and a high-fat content of the diet in the causation of alcoholic pancreatitis in humans.

Topics: Alcoholism; Amylases; Animals; Chymotrypsin; Chymotrypsinogen; Dietary Fats; Female; Lipase; Pancreas; Rats; Rats, Inbred Strains; Trypsin; Trypsin Inhibitors; Trypsinogen

The effects of sustained, high blood alcohol levels (216 +/- 120 mg/100 ml, S.D.) for 30 days on cholecystokinin (CCK)-mediated pancreatic exocrine function were studied in a rat model that achieves both maximally controlled, optimal nutrition and high alcohol intake (approximately 40.5% of total calories). In alcohol-fed rats, basal plasma levels of immunoreactive cationic trypsinogen (ICT) were reduced by 50% (P less than 0.05), but intravenous doses (0-30 IDU/kg/hr; 1 IDU = approximately 62.5 ng CCK-8) of cholecystokinin octapeptide (CCK-8) resulted in a 3-fold greater maximal concentration of ICT and an 80% steeper slope of the dose-response curve compared to those of pair-fed control animals. Basal plasma levels of amylase were not different in the two groups at basal conditions and did not change significantly following CCK-8 administration. In vitro studies with isolated pancreatic acini have shown that basal secretion of ICT into the media was similar in the two groups. However, ICT secretion in response to CCK-8 (30-3000 pM) was 2-fold greater in alcohol-fed rats than in pair-fed controls, resulting in a CCK-8 EC50 which was about half that of controls. On the contrary, the basal and maximal amylase secretion from acini isolated from alcohol-fed rats was reduced by 67 and 43%, respectively, causing a reduction in the magnitude of the response curve with almost identical EC50 and slopes. Despite the marked alterations in CCK-stimulated enzyme secretion, radioiodinated CCK-33 binding to receptors on acini isolated from both control and alcohol-fed rats was similar. Cellular concentrations of ICT and amylase, however, revealed similar patterns of alterations: 2 to 3-fold increase in ICT and 70% reduction in amylase in alcoholic acini compared to controls. These results indicate that the inverse changes in amylase and ICT secretions following continuous ethanol administration are probably due to differential effects on enzyme synthesis.

Topics: Alcoholism; Animals; Cholecystokinin; Ethanol; Male; Pancreas; Rats; Rats, Inbred Strains; Secretin; Sincalide; Time Factors; Trypsinogen

The effect of chronic alcohol abuse on the secretion of pancreatic exocrine proteins was studied. Pure pancreatic juice (PPJ) was obtained by endoscopic cannulation of the pancreatic duct from 21 healthy, nonalcoholic volunteers and 25 chronic alcoholics. Peak concentration and output of total proteins after sequential stimulation with secretin and cholecystokinin was elevated significantly in chronic alcoholics when compared to nonalcoholic subjects. The most striking change in the secretory proteins investigated was exhibited by the trypsinogens. Although the concentrations of all three trypsinogen variants were elevated significantly in PPJ of chronic alcoholics, most of the increase resulted from an approximately fivefold increase of the anionic variant, suggesting nonparallel alterations in the synthesis of pancreatic exocrine proteins. Whereas the ratio of cationic-anionic trypsinogen in the control group was consistently greater than one, it was, without exception, below one in the chronic alcoholics group. As there was no significant increase in trypsin inhibitor in PPJ of alcoholics, the ratio of trypsinogen-trypsin inhibitor showed a highly significant increase in this group. This distortion of the normal ratio in favor of trypsinogen may facilitate premature activation of pancreatic zymogens as postulated in acute pancreatitis. The concentrations of other zymogens and lysosomal hydrolases in PPJ of chronic alcoholics showed small, but not significant, increases, with the exception of leucine naphthylamidase which was significantly elevated. Nonparallel secretion of some exocrine proteins previously described in healthy nonalcoholic subjects was affected selectively by chronic ethanol ingestion. Thus, in chronic alcoholics the secretory kinetics of trypsinogen and chymotrypsinogen were altered, but trypsin inhibitor secretion remained apparently unaffected.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adult; Aged; Alcoholism; Cholecystokinin; Female; Humans; Hydrolases; Kinetics; Leucyl Aminopeptidase; Male; Middle Aged; Pancreatic Juice; Proteins; Secretin; Trypsin Inhibitors; Trypsinogen

The aim of the present study was to determine the effect of prolonged ethanol intake on the morphology and protein metabolism in the rat pancreatic acinar cells. Weight-matched triplets of Sprague-Dawley rats were fed Lieber-DeCarli diet containing 5% (wt/vol) concentration of ethanol, isocaloric amounts of Lieber-DeCarli diet, or rat chow ad libitum for 6, 12, and 18 mo. In the ethanol-fed group, histologic studies by light microscopy showed absence of protein plugs in the pancreatic ducts and/or pancreatitis, but electron-microscopic evaluation revealed progressive accumulation of lipid droplets in acinar and ductal cells. No definite changes in the mitochondria and endoplasmic reticulum were noticed. Biochemical studies revealed increased specific activity of trypsinogen, chymotrypsinogen, and lipase, and decreased specific activity of amylase. Trypsin-inhibiting capacity was decreased in the tissue and in the medium in a progressive fashion. There was no increase in the secretion of total protein. These data show a complex and a nonparallel alteration of specific digestive enzymes and trypsin inhibitor in this model of chronic ethanol intoxication that may be of relevance to occurrence of pancreatitis.

Topics: Alcoholism; Amylases; Animals; Chymotrypsinogen; Disease Models, Animal; Ethanol; Humans; Lipase; Male; Pancreas; Pancreatitis; Proteins; Rats; Rats, Inbred Strains; Trypsin Inhibitors; Trypsinogen

Polyacrylamide gel electrophoresis of pure pancreatic juice from 14 healthy normal subjects, 11 chronic alcoholics without detectable pancreatic disease, 15 patients with pancreatitis, and two with cancer of the pancreas consistently demonstrated the presence of two variants of trypsinogen with different electrophoretic mobilities. In healthy normal subjects the proportion of cationic to anionic trypsinogen was invariably greater than 1 and averaged about 2. In chronic alcoholics, patients with pancreatitis or cancer of the pancreas, this ratio, with a single exception, was below one and averaged about 0.45. The extraordinary consistency of these findings suggests that the quantitative relationship between cationic and anionic trypsinogen in human pancreatic juice may be a very sensitive indicator of incipient or existing pancreatic pathology. The most acceptable explanation for the reversal of the normal zymogen ratio in pancreatic disease is a selective increase in the synthesis of the anionic variant relative to that of the cationic species. Total trypsinogen concentrations differed widely from one another in the three patient groups, but the ratio of cationic to anionic trypsinogen exhibited little change and remained below 1. Our results also demonstrate for the first time a specific effect of chronic alcohol abuse on the secretory profile of a pancreatic enzyme in human subjects. A newly discovered minor, trypsinogen-like component of human pancreatic juice was found to be significantly increased in pancreatic juice of chronic alcoholics, decreased in pancreatic secretions of patients with pancreatitis, and barely detectable in those of two patients with cancer of the pancreas.

Topics: Adult; Alcoholism; Electrophoresis, Polyacrylamide Gel; Female; Humans; Male; Middle Aged; Pancreatic Juice; Pancreatic Neoplasms; Pancreatitis; Trypsinogen

Studies have been performed on pure pancreatic juice obtained by direct cannulation of the pancreatic duct in 2 patients with acute pancreatitis. The striking abnormalities observed, which were in marked contrast to our observations in 15 normal subjects, were high concentrations of protein throughout the period of secretin stimulation and the sporadic appearance of free proteolytic activity in many 1-min specimens throughout the collection period. In 1 subject repeat studies were performed after resolution of the pancreatitis when the profile observed was normal. Our findings are consistent with the hypothesis that obstruction of ductules and intraductal activation of zymogens may be important in the pathogenesis of acute pancreatitis.

Topics: Acute Disease; Adult; Alcoholism; Cholecystokinin; Chymotrypsinogen; Female; Humans; Male; Pancreatic Juice; Pancreatitis; Proteins; Secretin; Trypsin Inhibitors; Trypsinogen

Topics: Alanine Transaminase; Alcoholism; Amylases; Animals; Aspartate Aminotransferases; Chymotrypsin; Chymotrypsinogen; Humans; Male; Pancreas; Rats; Time Factors; Trypsin; Trypsin Inhibitors; Trypsinogen

Topics: Alcoholism; Humans; Pancreatic Juice; Pancreatitis; Trypsin; Trypsin Inhibitors; Trypsinogen

Topics: Alcoholism; Amylases; Animals; Dietary Carbohydrates; Dietary Fats; Dietary Proteins; Ethanol; Humans; Lipase; Male; Pancreas; Pancreatic Juice; Pancreatitis; Rats; Trypsinogen