Alcohols enhance caerulein-induced zymogen activation in pancreatic acinar cells. (41/463)

Activation of zymogens within the pancreatic acinar cell is an early feature of acute pancreatitis. Supraphysiological concentrations of cholecystokinin (CCK) cause zymogen activation and pancreatitis. The effects of the CCK analog, caerulein, and alcohol on trypsin and chymotrypsin activation in isolated pancreatic acini were examined. Caerulein increased markers of zymogen activation in a time- and concentration-dependent manner. Notably, trypsin activity reached a peak value within 30 min, then diminished with time, whereas chymotrypsin activity increased with time. Ethanol (35 mM) sensitized the acinar cells to the effects of caerulein (10(-10) to 10(-7) M) on zymogen activation but had no effect alone. The effects of ethanol were concentration dependent. Alcohols with a chain length of >or=2 also sensitized the acinar cell to caerulein; the most potent was butanol. Branched alcohols (2-propanol and 2-butanol) were less potent than aliphatic alcohols (1-propanol and 1-butanol). The structure of an alcohol is related to its ability to sensitize acinar cells to the effects of caerulein on zymogen activation.  (+info)

Pharmacological studies of caerulein. II. The possibility of mediation through the central nervous system. (42/463)

With low doses of caerulein given intravenously or subcutaneously, vomiting could be induced. Although vomiting was not inhibited by chlorpromazine or atropine, a strong inhibition was evident when metoclopramide was administered subcutaneously at such doses as inhibit vomiting induced by oral administration of CuSO4. Even high doses of caerulein failed to induced vomiting in vagotomized and splanchnicotomized dogs. With intraventricular injection, no effects were observed on blood pressure, respiration or gastrointestinal motility, and vomiting was not induced. Therefore, a reflex mechanism appears to be involved in vomiting induced by caerulein. It is suggested that the actions of caerulein may not be mediated through the central nervous system.  (+info)

Biosynthesis of caerulein in the skin of Xenopus laevis: partial sequences of precursors as deduced from cDNA clones. (43/463)

The decapeptide caerulein represents one of the main constituents of the skin secretion of Xenopus laevis. Total mRNA was isolated from skin, transcribed into cDNA and inserted via GC-tailing into the plasmid pUC8. Among the transformants, 300 clones were selected at random and screened with a cDNA primed with the synthetic deoxynucleotide d(AGTCCATCCA), which is complementary to the mRNA region coding for the fragment Trp-Met-Asp-Phe of cerulein. Of nine strongly hybridizing clones, three were sequenced and these were found to contain inserts with very similar nucleotide sequences. The cloned cDNAs code for parts of two different caerulein precursors. These contain one or two copies of caerulein and five additional amino acids located between pairs of arginine residues. The extra glycine at the carboxy terminus is considered to serve as the signal for amidation, while the tetrapeptide Phe-Ala-Asp-Gly, linked to the amino end of caerulein in these precursors, must be cleaved by an unusual processing mechanism.  (+info)

Sensory nerves in central and peripheral control of pancreatic integrity by leptin and melatonin. (44/463)

Central nervous system affects pancreatic secretion of enzymes however, the neural modulation of acute pancreatitis has not been investigated. Leptin and melatonin have been recently reported to affect the inflammatory response of various tissues. The identification of specific receptors for both peptides in the pancreas suggests that leptin and melatonin could contribute to the pancreatic protection against inflammation. The aim of this study was: 1/ to compare the effect of intracerebroventricular (i.c.v.) or intraperitoneal (i.p.) administration of leptin or melatonin on the course of caerulein-induced pancreatitis (CIP) in the rat, 2/ to examine the involvement of sensory nerves (SN) and calcitonin gene-related peptide (CGRP) in pancreatic protection afforded by leptin or melatonin, 3/ to assess the effect of tested peptides on lipid peroxidation products (MDA + 4-HNE) in the pancreas of CIP rats, 4/ to investigate the influence of leptin or melatonin on nitric oxide (NO) release from isolated pancreatic acini and 5/ to determine the effects of caerulein and leptin on leptin receptor gene expression in these acini by RT-PCR. CIP was induced by subcutaneous (s.c.) infusion of caerulein (25 microg/kg) to the conscious rats, confirmed by the significant increases of pancreatic weight and plasma amylase and by histological examination. This was accompanied in marked reduction of pancreatic blood flow and significant rise of MDA + 4-HNE in the pancreas. Leptin or melatonin were administered i.p. or i.c.v. 30 min prior to the start of CIP. Deactivation of SN was produced by s.c. capsaicin (100 mg/kg). An antagonist of CGRP, CGRP 8-37 (100 microg/kg i.p.), was given together with leptin or melatonin to the CIP rats. MDA + 4-HNE was measured using LPO commercial kit. NO was determined using the Griess reaction. Pretreatment of CIP rats with i.p. leptin (2 or 10 microg/kg) or melatonin (10 or 50 mg/kg) significantly attenuated the severity of CIP. Similar protective effects were observed following i.c.v. application of leptin (0.4 or 2 microg/rat) but not melatonin (10 or 40 microg/rat) to the CIP rats. Capsaicin deactivation of SN oradministration of CGRP 8-37 abolished above beneficial effects of leptin on CIP, whereas melatonin-induced protection of pancreas was unaffected. Pretreatment with i.p. melatonin (10 or 50 mg/kg), but not leptin, significantly reduced MDA + 4-HNE in the pancreas of CIP rats. Leptin (10(-10) - 10(-6) M) but not melatonin (10(-8) - 10(-5) M) significantly stimulated NO release from isolated pancreatic acini. Leptin receptor gene expression in these acini was significantly increased by caerulein and leptin. We conclude that 1/ central or peripheral pretreatment with leptin protects the pancreas against its damage induced by CIP, whereas melatonin exerts its protective effect only when given i.p., but not following its i.c.v. adminstration, 2/ activation of leptin receptor in the pancreatic acini appears to be involved in the beneficial effects of leptin on acute pancreatitis, 3/ the protective effects of leptin involve sensory nerves, CGRP and increased generation of NO whereas melatonin-induced protection of the pancreas depends mainly on the antioxidant local effect of this indole, and scavenging of the radical oxygen species in the pancreatic tissue.  (+info)

Calcium and pancreatic secretion-dynamics of subcellur calcium pools in resting and stimulated acinar cells. (45/463)

1 Pulse-chase experiments were carried out on pancreatic tissue lobules incubated in vitro, with 45Ca as the tracer, in order to shed some light on the functional significance of the calcium pools associated with the various cell organelles of the acinar cell, especially in relation to stimulus-secretion coupling. 2 The kinetics of tracer uptake and release which were observed in the intact lobules suggest the existence of a number of intracellular pools, whose rate of exchange is slower than that across teh plasmalemma. 3 The various subcellular fractions accumulate the tracer in different amounts: some (rough microsomes and postmicrosomal supernatant) showed little radioactivity and some (smooth microsomes and zymogen granule membranes) were heavily labelled; mitochondria and zymogen granules showed intermediate values. 4 The fractions are heterogeneous also in relation to the time course of uptake and release of the tracer: in rough and smooth microsomes and, especially, in the postmicrosomal supernatant both rates were fast; zymogen granules and zymogen granule membranes showed slow rates of uptake and little release during chase; intermediate rates were found in mitochondria. 5 In agreement with previous findings we observed that in 45Ca preloaded lobules, stimulation of secretion (brought about by the secretagogue polypeptide caerulein) results in an increase of the tracer release which seems to be due primarily to the rise of the intracellular concentration of free Ca2+ and to the consequent increase of the transmembrane Ca2+ efflux. Among the cell fractions isolated from stimulated lobules only the mitochondria exhibited a significantly lower 45Ca level relative to the unstimulated controls. 6 It is concluded that, of the organelle-bound calcium pools, that associated with the mitochondria might be involved in the regulation of the calcium-dependent functions, including stimulus-secretion coupling; the calcium associated with the zymogen granule content probably has a role in the architecture of the organelle and in the functionality of the pancreatic juice, while the calcium bound to the membrane of the granules might be concerned with the regulation of its permeability properties.  (+info)

Secretion of electrolytes by the pancreas of the anaestetized rat. (46/463)

1. HCO-3, Na+ and K+ concentrations were measured in bile-free pancreatic juice collected from fasted and fed anaesthetized rats. 2. Resting flow rates averaged 0.62 mul. g-1 .min-1 (fasted) and 2.8 mul. g-1. min-1 (fed) and the mean HCO-3 concentrations, respectively, were 25.8 and 33.3 mM. 3. In fasted rats, instillation of HCl into the duodenum caused flow rate to increase threefold and HCO-3 concentrations to double (66 mM). Intravenous infusion of pure natural (GIH) secretin caused a fivefold increase in flow rate; HCO-3 concentrations, again, doubled (67.5 mM). Infusion of synthetic secretin produced effects essentially the same as those produced by GIH secretin. 4. Infusion of Boots secretin caused a thirteenfold increase in flow rate (8.32 mul.g-1. min-1) but HCO-3 concentrations rose only slightly (43.3 mM). However, following cessation of infusion, when flow rate approximated the maximum obtained with pure secretin, the HCO-3 concentration was much higher (57.2 mM at 3.19 uml.g-1.min-1). In fed animals the responses were similar but maximum flow rates were greater (12 mul. g-1. min-1). 5. Infusion of caerulein produced a secretory rate slightly less than with Boots secretin (5.06 mul. g-1.min-1) and HCO-3 concentrations were plasmalike (30.2 mM); infusion of the synthetic octapeptide of cholecystokinin (OP-CCK) gave similar flow rates and HCO-3 concentrations. 6. Infusion of a mixture of caerulein and GIH secretin mimicked closely the effect of Boots secretin. At maximum flow rates (7.6 mul. g-1. min-1) the HCO-3 concentration was 43.7 mM and at lower flow rates (3.90 mul.g-1. min-1) it rose to 54.2mM. 7. It is concluded that the response of the rat pancreas to secretin is qualitatively similar to that of all other vertebrates so far studied, but, relative to other animals, the response is sluggish. In contrast, the rat pancreas responds well to cholecystokinin (CCK) stimulation, yielding a juice with plasma-like HCO-3 concentration. Boots secretin, which is heavily contaminated with CCK, causes a mixed response resembling that of CCK at high secretory rates and that of pure secretin at lower rates. 8. An unexplained feature of rat pancreatic juice was that K+ concentrations, although plasma-like in unstimulated samples, rose to about 8mM when flow rate increases as a result of secretin, but not CCK, stimulation. In all other animals so far studied, the K+ concentration has been found to be independent of flow rate.  (+info)

The influence of cholecystokinin on gastric myoelectrical activity in duodenal ulcer following Helicobacter pylori eradication--an electrogastrographic study. (47/463)

Cholecystokinin (CCK) plays an important role in the regulation of postprandial gastric motor activity which was found to be abnormal in duodenal ulcer patients. This study was designed to compare the influence of CCK on gastric myoelectrical function in duodenal ulcer patients and healthy controls. Fifteen patients with active duodenal ulcer and Helicobacterpylori (H. pylori) infection and 15 healthy controls were included into this study. Electrogastrography (EGG) was performed before and 4 weeks after the eradication of H. pylori in ulcer patients and in healthy controls. We compared EGG parameters in the fasting and postprandial period and during intravenous infusion of caerulein, an analog of CCK with or without addition of loxiglumide, a specific CCK-1 receptor antagonist. The amplitude of fasting EGG in duodenal ulcer patients was similar to that in control subjects and was not affected by H. pylori eradication. In contrast, the amplitude of postprandial EGG was markedly increased in duodenal ulcer patients when compared to that in healthy controls and it was significantly reduced following the eradication of H. pylori. The blockade of CCK-1 receptors with loxiglumide in healthy controls or H. pylori eradicated ulcer patients significantly enhanced postprandial EGG amplitude almost to the level observed in the infected duodenal ulcer patients, but failed to affect this amplitude in ulcer patients. Exogenous caerulein, an analog of CCK, failed to affect EGG amplitude in duodenal ulcer patients with H. pylori infection, but it reduced significantly EGG amplitude in these patients after H. pylori eradication and in control subjects. This inhibitory effect of caerulein in H. pylori negative ulcer patients and healthy controls was abolished by the addition of loxiglumide. Ulcer patients showed significant dysrhythmia with tachygastria up to 20% of the recording time both under basal conditions and postprandially and H. pylori eradication was followed by a significant decrease in tachygastria to about 5%, the value being similar to that in healthy controls. We conclude that the amplitude and frequency of gastric myoelectrical activity are enhanced in duodenal ulcer patients and impaired in response to CCK but these changes can be normalized by successful H. pylori eradication.  (+info)

Different modes of NF-kappaB/Rel activation in pancreatic lobules. (48/463)

The eukaryotic transcription factor nuclear factor-kappaB (NF-kappaB)/Rel is activated by a large variety of stimuli. It has been demonstrated that NF-kappaB/Rel is induced during the course of cerulein pancreatitis. Here, we show that NF-kappaB/Rel is differentially activated in pancreatic lobules. Cerulein induces NF-kappaB/Rel via activation of IkappaB kinase (IKK), which causes degradation of IkappaBalpha but not IkappaBbeta. Tumor necrosis factor-alpha-mediated IKK activation leads to IkappaBalpha and IkappaBbeta degradation. In contrast, oxidative stress induced by H(2)O(2) activates NF-kappaB/Rel independent of IKK activation and IkappaBalpha degradation; instead IkappaBalpha is phosphorylated on tyrosine. H(2)O(2) but not cerulein-mediated NF-kappaB/Rel activation can be blocked by stabilizing microtubules with Taxol. Inhibition of tubulin polymerization with nocodazole causes NF-kappaB/Rel activation in pancreatic lobules. These results propose three different pathways of NF-kappaB/Rel activation in pancreatic acinar cells. Furthermore, these data demonstrate that microtubules play a key role in IKK-independent NF-kappaB/Rel activation following oxidative stress.  (+info)