RESUMO
Peripheral administration of cholecystokinin (CCK)-8 or secretin activates oxytocin (OXT)-secreting neurons in the hypothalamus. Although OXT is involved in the regulation of feeding behavior, detailed mechanism remains unclear. In the present study, we examined the central OXTergic pathways after intraperitoneally (i.p.) administration of CCK-8 and secretin using male OXT-monomeric red fluorescent protein 1 (mRFP1) transgenic rats and male Wistar rats. I.p. administration of CCK-8 (50µg/kg) and secretin (100µg/kg) decreased food intake in these rats. While i.p. administration of CCK-8 decreased water intake, i.p. administration of secretin increased water intake. Immunohistochemical study revealed that Fos-Like-Immunoreactive cells were observed abundantly in the brainstem and in the OXT neurons in the dorsal division of the parvocellular paraventricular nucleus (dpPVN). We could observe marked increase of mRFP1 fluorescence, as an indicator for OXT, in the dpPVN and mRFP1-positive granules in axon terminals of the dpPVN OXT neurons in the nucleus tractus solitarius (NTS) after i.p. administration of CCK-8 and secretin. These results provide us the evidence that, at least in part, i.p. administration of CCK-8 or secretin might be involved in the regulation of feeding/drinking via a OXTergic pathway from the dpPVN to the NTS.
Assuntos
Comportamento de Ingestão de Líquido/efeitos dos fármacos , Comportamento Alimentar/efeitos dos fármacos , Proteínas Luminescentes/metabolismo , Ocitocina/metabolismo , Secretina/farmacologia , Sincalida/farmacologia , Animais , Tronco Encefálico/metabolismo , Colecistocinina , Fluorescência , Hipotálamo/metabolismo , Injeções Intraperitoneais , Masculino , Neurônios/metabolismo , Terminações Pré-Sinápticas/metabolismo , Ratos Transgênicos , Ratos Wistar , Proteína Vermelha FluorescenteRESUMO
Secretin and its receptors show wide distribution in the central nervous system. It was demonstrated previously that intravenous (i.v.) and intracerebroventricular (i.c.v.) application of secretin influenced the behavior of rat, mouse, and human. In our previous experiment, we used a special animal model, Japanese waltzing mice (JWM). These animals run around without stopping (the ambulation distance is very limited) and they do not bother with their environment. The i.c.v. secretin attenuated this hyperactive repetitive movement. In the present work, the effect of i.c.v. and intranasal (i.n.) application of secretin was compared. We have also looked for the presence of secretin receptors in the brain structures related to motor functions. Two micrograms of i.c.v. secretin improved the horizontal movement of JWM, enhancing the ambulation distance. It was nearly threefold higher in treated than in control animals. The i.n. application of secretin to the left nostril once or twice a day or once for 3 days more effectively enhanced the ambulation distance than i.c.v. administration. When secretin was given twice a day for 3 days it had no effect. Secretin did not improve the explorative behavior (the rearing), of JWM. With the use of in situ hybridization, we have found very dense secretin receptor labeling in the cerebellum. In the primary motor cortex and in the striatum, only a few labeled cells were seen. It was supposed that secretin exerted its effect through specific receptors, mainly present in the cerebellum.
Assuntos
Comportamento Exploratório/efeitos dos fármacos , Atividade Motora/efeitos dos fármacos , Secretina/farmacologia , Administração Intranasal , Animais , Cerebelo/química , Cerebelo/efeitos dos fármacos , Corpo Estriado/química , Avaliação Pré-Clínica de Medicamentos , Feminino , Hipercinese/tratamento farmacológico , Hipercinese/genética , Hibridização In Situ , Injeções Intraventriculares , Masculino , Camundongos , Camundongos Mutantes Neurológicos , Atividade Motora/fisiologia , Córtex Motor/química , Proteínas do Tecido Nervoso/agonistas , Proteínas do Tecido Nervoso/análise , Proteínas do Tecido Nervoso/fisiologia , Ratos , Ratos Sprague-Dawley , Receptores Acoplados a Proteínas G/agonistas , Receptores Acoplados a Proteínas G/análise , Receptores Acoplados a Proteínas G/fisiologia , Receptores dos Hormônios Gastrointestinais/agonistas , Receptores dos Hormônios Gastrointestinais/análise , Receptores dos Hormônios Gastrointestinais/fisiologia , Secretina/administração & dosagem , Secretina/uso terapêuticoRESUMO
Fluid balance is critical to life and hence is tightly controlled in the body. Angiotensin II (ANGII), one of the most important components of this regulatory system, is recognized as a dipsogenic hormone that stimulates vasopressin (VP) expression and release. However, detailed mechanisms regarding how ANGII brings about these changes are not fully understood. In the present study, we show initially that the osmoregulatory functions of secretin (SCT) in the brain are similar to those of ANGII in mice and, more important, we discovered the role of SCT as the link between ANGII and its downstream effects. This was substantiated by the use of two knockout mice, SCTR(-/-) and SCT(-/-), in which we show the absence of an intact SCT/secretin receptor (SCTR) axis resulted in an abolishment or much reduced ANGII osmoregulatory functions. By immunohistochemical staining and in situ hybridization, the proteins and transcripts of SCT and its receptor are found in the paraventricular nucleus (PVN) and lamina terminalis. We propose that SCT produced in the circumventricular organs is transported and released in the PVN to stimulate vasopressin expression and release. In summary, our findings identify SCT and SCTR as novel elements of the ANGII osmoregulatory pathway in maintaining fluid balance in the body.
Assuntos
Angiotensina II/farmacologia , Secretina/metabolismo , Secretina/farmacologia , Animais , Ingestão de Líquidos/efeitos dos fármacos , Feminino , Hipotálamo/efeitos dos fármacos , Hipotálamo/metabolismo , Imuno-Histoquímica , Hibridização In Situ , Camundongos , Camundongos Knockout , Núcleo Hipotalâmico Paraventricular/efeitos dos fármacos , Núcleo Hipotalâmico Paraventricular/metabolismo , Hipófise/efeitos dos fármacos , Hipófise/metabolismo , Reação em Cadeia da Polimerase , Receptores Acoplados a Proteínas G/genética , Receptores Acoplados a Proteínas G/metabolismo , Receptores dos Hormônios Gastrointestinais/genética , Receptores dos Hormônios Gastrointestinais/metabolismo , Secretina/genética , Vasopressinas/metabolismo , Equilíbrio Hidroeletrolítico/efeitos dos fármacosRESUMO
BACKGROUND: Secretin is a "gut-brain" peptide whose neural function is as yet poorly understood. Several clinical studies have reported modestly increased social interaction in autistic children following intravenous secretin administration. Very recently secretin also was administered to schizophrenic patients and found to increase social interaction in some individuals. METHODS: In light of this finding, we assessed the ability of secretin to reverse phencyclidine- (PCP) induced impairment in prepulse inhibition (PPI), a leading animal model of sensorimotor gating deficits in schizophrenia. RESULTS: Similar to atypical antipsychotics, secretin (1, 3, 10, 30, and 100 microg/kg) partially and dose-dependently reversed the PCP-induced deficit in PPI without significantly affecting baseline startle when administered intraperitoneally (IP) 10 minutes following IP administration of PCP (3 mg/kg). CONCLUSIONS: This finding may be relevant to observations of antipsychotic efficacy of secretin in schizophrenic patients as well as our previous report that systemically administered secretin is capable of modulating conditioned fear, even at quite low doses.
Assuntos
Comportamento Animal/efeitos dos fármacos , Antagonistas de Aminoácidos Excitatórios/farmacologia , Fenciclidina/farmacologia , Reflexo de Sobressalto/efeitos dos fármacos , Secretina/farmacologia , Estimulação Acústica , Animais , Percepção Auditiva/efeitos dos fármacos , Condicionamento Clássico/efeitos dos fármacos , Condicionamento Clássico/fisiologia , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Interações Medicamentosas , Medo/efeitos dos fármacos , Medo/fisiologia , Humanos , Masculino , Ratos , Ratos Sprague-Dawley , Esquizofrenia/tratamento farmacológico , Psicologia do Esquizofrênico , Secretina/administração & dosagem , Secretina/uso terapêuticoRESUMO
Recent research points to the connection between behavioral and gut disorders. Early adverse events are associated with inflammatory bowel disease (IBD). In animal models, maternal deprivation and social isolation predispose to gastric erosion and brain pathology. This study examined (1) brain effects of chronic gastrointestinal inflammation in a rat model of acquired IBD and (2) whether such changes are resolved by individual secretin (S) or oxytocin (OT) peptide treatment. Neurological manifestations of IBD were mapped by c-fos gene expression in male Sprague-Dawley rats (n = 10) with trinitrobenzene sulfonic acid (TNBS)-induced IBD vs controls (n = 11). IBD was characterized by moderate/severe infiltration of inflammatory cells 10 d after TNBS infusion. Age-matched pairs were processed for immunocytochemical detection of Fos, expressed when neurons are stimulated. S or OT (100 mg/250 mL saline) or equivolume saline was administered iv by Alzet pump for 20 d after disease onset. Degree of resolution of colitis-induced brain activation was assessed by c-fos expression, and mean numbers of Fos-immunoreactive nuclei for each group were compared using Independent Samples T-test. Chronic IBD activated periventricular gray, hypothalamic/visceral thalamic stress axes and cortical domains, and septal/preoptic/amygdala, brain areas abnormal in autism. Single peptide treatment with S or OT did not alter the effects of inflammation on the brain. Brain areas concomitantly activated by visceral inflammation are those often abnormal in autism, suggesting that IBD could be a model for testing treatments of autism. Other single and combined peptide treatments of IBD should be tested. The clinical implications for treating autism, IBD, and concomitant sickness behaviors with peptide therapy, with or without maternal nurturing as a natural equivalent, are presented.
Assuntos
Transtorno Autístico/tratamento farmacológico , Transtorno Autístico/fisiopatologia , Doenças Inflamatórias Intestinais/tratamento farmacológico , Doenças Inflamatórias Intestinais/fisiopatologia , Ocitocina/farmacologia , Secretina/farmacologia , Tonsila do Cerebelo/efeitos dos fármacos , Tonsila do Cerebelo/fisiopatologia , Animais , Transtorno Autístico/complicações , Doença Crônica , Modelos Animais de Doenças , Hipotálamo/efeitos dos fármacos , Hipotálamo/fisiopatologia , Doenças Inflamatórias Intestinais/complicações , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
Acetylcholine potentiates secretin-stimulated ductal secretion by Ca(2+)-calcineurin-mediated modulation of adenylyl cyclase. D2 dopaminergic receptor agonists inhibit secretin-stimulated ductal secretion via activation of protein kinase C (PKC)-gamma. No information exists regarding the effect of adrenergic receptor agonists on ductal secretion in a model of cholestasis induced by bile duct ligation (BDL). We evaluated the expression of alpha-1A/1C, -1beta and beta-1 adrenergic receptors in liver sections and cholangiocytes from normal and BDL rats. We evaluated the effects of the alpha-1 and beta-1 adrenergic receptor agonists (phenylephrine and dobutamine, respectively) on bile and bicarbonate secretion and cholangiocyte IP(3) and Ca(2+) levels in normal and BDL rats. We measured the effect of phenylephrine on lumen expansion in intrahepatic bile duct units (IBDUs) and cyclic adenosine monophosphate (cAMP) levels in cholangiocytes from BDL rats in the absence or presence of BAPTA/AM and Gö6976 (a PKC-alpha inhibitor). We evaluated if the effects of phenylephrine on ductal secretion were associated with translocation of PKC isoforms leading to increased protein kinase A activity. Alpha-1 and beta-1 adrenergic receptors were present mostly in the basolateral domain of cholangiocytes and, following BDL, their expression increased. Phenylephrine, but not dobutamine, increased secretin-stimulated choleresis in BDL rats. Phenylephrine did not alter basal but increased secretin-stimulated IBDU lumen expansion and cAMP levels, which were blocked by BAPTA/AM and Go6976. Phenylephrine increased IP(3) and Ca(2+) levels and activated PKC-alpha and PKC-beta-II. In conclusion, coordinated regulation of ductal secretion by secretin (through cAMP) and adrenergic receptor agonist activation (through Ca(2+)/PKC) induces maximal ductal bicarbonate secretion in liver diseases. (Supplementary material for this article can be found on the HEPATOLOGY website (http://interscience.wiley.com/jpages/0270-9139/suppmat/index.html).
Assuntos
Agonistas de Receptores Adrenérgicos alfa 1 , Ductos Biliares/metabolismo , Cálcio/fisiologia , AMP Cíclico/metabolismo , Proteína Quinase C/fisiologia , Agonistas alfa-Adrenérgicos/farmacologia , Animais , Bicarbonatos/metabolismo , Bile/metabolismo , Ductos Biliares/citologia , Ductos Biliares Intra-Hepáticos/metabolismo , Transporte Biológico , Membrana Celular , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Sinergismo Farmacológico , Isoenzimas/metabolismo , Ligadura , Masculino , Fenilefrina/farmacologia , Isoformas de Proteínas/metabolismo , Proteína Quinase C/metabolismo , Ratos , Ratos Endogâmicos F344 , Receptores Adrenérgicos alfa 1/metabolismo , Secretina/farmacologia , Regulação para CimaRESUMO
The aim of the study reported here was to investigate changes in the digestive enzyme content in the pancreas after food and secretagogue stimulation. Rats from which food had been withheld overnight were either fed (between 6 and 8 a.m.) or not before euthanasia and pancreatic excision (at 8 a.m.: 21 not fed and 21 fed) and at 4 (12 p.m.: six not fed and six fed) and 8 h later (4 p.m.: six not fed and six fed). Another 16 rats were anesthetized, fitted with jugular vein and pancreatic duct catheters, and infused with the secretagogues, CCK-33 and secretin, during 1.5 h of pancreatic juice collection before euthanasia and pancreatic excision. The pancreata were homogenized, and total soluble protein and individual enzyme (trypsin and amylase) tissue contents were analyzed. Results indicated lower amounts of protein and enzymes remaining in the pancreata of the fed, compared with non-fed rats. Enzyme values indicated recovery within four hours in fed rats, but non-fed rats also had increased values during daytime. High enzyme secretion during the high dose of hormonal stimulation was reflected in lower enzyme values remaining in the pancreas, compared with that in response to low-dose stimulation. Results indicated that stimulation of the pancreas, either by food ingestion or exogenous secretagogues, lowers the amounts of digestive enzymes remaining in the pancreas, and imply that stimulation and circadian rhythms influence the pancreatic enzyme content at euthanasia. This finding should be borne in mind in interpretation of data from pancreatic studies.
Assuntos
Amilases/metabolismo , Ingestão de Alimentos/fisiologia , Pâncreas/enzimologia , Tripsina/metabolismo , Animais , Colecistocinina/administração & dosagem , Colecistocinina/farmacologia , Relação Dose-Resposta a Droga , Privação de Alimentos , Fármacos Gastrointestinais/farmacologia , Infusões Intravenosas , Masculino , Pâncreas/efeitos dos fármacos , Pâncreas/metabolismo , Suco Pancreático/efeitos dos fármacos , Suco Pancreático/enzimologia , Suco Pancreático/metabolismo , Proteínas/metabolismo , Ratos , Ratos Sprague-Dawley , Secretina/administração & dosagem , Secretina/farmacologiaRESUMO
Three distinct complementary DNAs for vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase-activating peptide (PACAP) receptors have been cloned and designated VIP-1 receptor (VIP-1R), VIP-2 receptor (VIP-2R), and PACAP receptor (PACAP-R). In the present study, we have characterized the binding sites on primary mouse calvarial osteoblasts for VIP and related peptides. By analyzing the cAMP response, the rank order of response observed was PACAP 38 > PACAP 27 > helodermin > VIP > helospectin > glucagon > PHI >>> secretin. The VIP-2R/PACAP-R antagonist, PACAP 6-38, inhibited both VIP- and PACAP-stimulated cAMP formation. Binding studies using an atomic force microscopy (AFM) technique showed high affinity binding for VIP and PACAP 38, but not for secretin. Radioligand binding studies using (125)I-VIP and (125)I-PACAP 38 demonstrated a more specific and higher affinity binding for PACAP 38 than for VIP. Secretin failed to inhibit both (125)I-VIP and (125)I-PACAP 38 binding. RT-PCR demonstrated that undifferentiated mouse calvarial osteoblasts express messenger RNA for VIP-2R, but not for VIP-1R or PACAP-R. When the osteoblasts were cultured for 20 days to induce bone noduli formation, VIP-1R, in addition to VIP-2R, were expressed when the nodules started to mineralize at 12 days. Taken together, these data demonstrate that mouse calvarial osteoblasts express functional VIP-2R with higher affinity binding for PACAP than for VIP and that the VIP-1R expression is induced during osteoblastic differentiation.
Assuntos
Neuropeptídeos/farmacologia , Osteoblastos/fisiologia , Receptores de Peptídeo Intestinal Vasoativo/fisiologia , Peptídeo Intestinal Vasoativo/farmacologia , Animais , Células Cultivadas , AMP Cíclico/metabolismo , Glucagon/farmacologia , Peptídeos e Proteínas de Sinalização Intercelular , Cinética , Camundongos , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Peptídeos/farmacologia , Polipeptídeo Hipofisário Ativador de Adenilato Ciclase , Reação em Cadeia da Polimerase , Ensaio Radioligante , Receptores de Peptídeo Intestinal Vasoativo/efeitos dos fármacos , Receptores de Peptídeo Intestinal Vasoativo/genética , Receptores Tipo II de Peptídeo Intestinal Vasoativo , Secretina/farmacologia , CrânioRESUMO
Six neuropeptides: short and long form of the pituitary adenylate cyclase activating polypeptide (PACAP), i.e. PACAP27 and PACAP38, vasoactive intestinal peptide (VIP), peptide histidine-isoleucine (PHI), secretin and glucagon, members of the secretin/VIP/PACAP superfamily ofpolypeptides, were tested for their ability to stimulate cyclic AMP formation in [3H]adenine-prelabeled slices of the chick hypothalamus and cerebral cortex. Of the tested peptides, only PACAP evoked pronounced and significant responses in the two analyzed brain structures. Although magnitude of the responses varied in different experiments, the effects of both forms of PACAP were usually larger in the cerebral cortex than in the hypothalamus. Glucagon, PHI (both used at concentrations 0.01-1 microM) and VIP (0.1-3 microM) induced concentration-dependent yet comparatively small effects that did not reach statistical significance, while secretin (0.1-3 microM) had no effect.
Assuntos
Córtex Cerebral/metabolismo , Galinhas/metabolismo , AMP Cíclico/biossíntese , Hipotálamo/metabolismo , Neuropeptídeos/farmacologia , Secretina/farmacologia , Peptídeo Intestinal Vasoativo/farmacologia , Sequência de Aminoácidos , Animais , Córtex Cerebral/efeitos dos fármacos , Glucagon/farmacologia , Hipotálamo/efeitos dos fármacos , Masculino , Dados de Sequência Molecular , Polipeptídeo Hipofisário Ativador de Adenilato CiclaseRESUMO
In order to establish the measurement of gastric mucin secreted from cultured mucous cells, rat gastric mucin was purified from secreted mucus with Sepharose CL-4B column chromatography. Gastric mucin was measured by dot blot analysis using an enzyme-linked lectin (soybean agglutinin) assay in a good concentration-dependent manner. Surface epithelial cells were dispersed by limited digestion of a rat everted stomach and collected by density gradient centrifugation with Percoll. These cells were inoculated onto gelled collagen dishes, then cultured in a medium supplemented with 10% fetal calf serum under a 5% CO2 atmosphere in air. Changing the medium after a 2-d culture, the cells were cultured for another 3 d. During the culture, the numbers of cells each day were almost equal, but mucin contents in the cells increased, and then dropped at day 5 after inoculation. At that time, the edge of the cell layer peeled off and the cells adhered to each other. Using 2-d cultured cells, the effects of some secretagogues on mucin secretion were investigated. Carbachol, secretin, CCK-8 and prostaglandin E2 (PGE2) strongly stimulated mucin secretion, and gastrin I weakly did. However, histamine offered no stimulation.
Assuntos
Mucinas Gástricas/metabolismo , Mucosa Gástrica/metabolismo , Animais , Carbacol/farmacologia , Células Cultivadas , Dinoprostona/farmacologia , Células Epiteliais , Epitélio/efeitos dos fármacos , Epitélio/metabolismo , Mucinas Gástricas/isolamento & purificação , Mucosa Gástrica/efeitos dos fármacos , Gastrinas/farmacologia , Histamina/farmacologia , Ratos , Ratos Wistar , Secretina/farmacologia , Sincalida/farmacologiaAssuntos
Glândulas Suprarrenais/efeitos dos fármacos , Hipotálamo/efeitos dos fármacos , Hipófise/efeitos dos fármacos , Secretina/farmacologia , Glândulas Suprarrenais/metabolismo , Hormônio Adrenocorticotrópico/sangue , Animais , Corticosterona/sangue , Corticosterona/metabolismo , Feminino , Hipotálamo/metabolismo , Hipófise/metabolismo , Ratos , Ratos WistarRESUMO
This study characterizes regional regulation of adenylate cyclase by galanin, neuropeptide Y (NPY), secretin and vasoactive intestinal peptide (VIP) in rat brain frontal cortex, hypothalamus and hippocampus. In our experimental system, galanin caused small detectable activation (10-20%) of basal adenylate cyclase activity in frontal cortex and hippocampus but had no effect on basal adenylate cyclase activity in hypothalamus. Galanin inhibited forskolin-stimulated adenylate cyclase in all three brain regions-hypothalamus, hippocampus and frontal cortex by 54.5%, 44.3% and 25.7%, respectively. NPY reduced basal and forskolin-stimulated enzyme activities by 35% only in frontal cortex, but not in the other two brain areas. Secretin had no effect in frontal cortex but caused similar adenylate cyclase activation in hypothalamus and hippocampus. VIP had a stimulatory effect of 32.8% and 32.4% in frontal cortex and hippocampus, respectively. The results indicate regional differences in adenylate cyclase modulation by the four peptides and reveal interesting relations in comparison with peptide and receptor densities in the three investigated brain regions.
Assuntos
Adenilil Ciclases/metabolismo , Lobo Frontal/fisiologia , Neuropeptídeo Y/farmacologia , Peptídeos/farmacologia , Adenilil Ciclases/efeitos dos fármacos , Animais , Galanina , Hipocampo/efeitos dos fármacos , Hipotálamo/efeitos dos fármacos , Masculino , Ratos , Secretina/farmacologia , Peptídeo Intestinal Vasoativo/farmacologiaRESUMO
Acute, subacute, and chronic experiments were performed to examine the toxicity, specific action, analgetic, and physicochemical properties of suicrepan derived from the porcine duodenal mucosa and the hormone secretin contained in it. Suicrepan was found to be an effective pancreatic extrasecretory function stimulant, by showing therapeutical benefits in experimental pancreatitis and analgesic effects. The drug was demonstrated to display a low toxicity.
Assuntos
Secretina/farmacologia , Doença Aguda , Analgésicos/farmacologia , Animais , Fenômenos Químicos , Físico-Química , Cães , Relação Dose-Resposta a Droga , Avaliação Pré-Clínica de Medicamentos , Masculino , Camundongos , Suco Pancreático/efeitos dos fármacos , Suco Pancreático/metabolismo , Pancreatite/tratamento farmacológico , Coelhos , Ratos , Secretina/química , Secretina/uso terapêutico , Secretina/toxicidade , Fatores de TempoRESUMO
The roles of Ca2+ in agonist-induced pepsinogen secretion from guinea pig chief cells remain unclear. We used cholecystokinin octapeptide (CCK-8) or secretin alone or with thapsigargin (TG) to clarify these roles. TG releases Ca2+ from intracellular stores by inhibiting microsomal Ca(2+)-adenosinetriphosphatase (ATPase), thereby depleting intracellular Ca2+ (Cai2+) stores. In most cells TG also causes Ca2+ influx. In the present study, with an extracellular Ca2+ concentration ([Ca2+]o) of 1.5 mM, CCK-8 (0.1 microM) caused a rapid increase in pepsinogen secretion; however, the rate decreased with time. With [Ca2+]o = 0, the initial increase was similar but later secretion was abolished, suggesting that Ca2+ influx was important for sustained secretion. With [Ca2+]o = 1.5 mM, TG (0.1 microM) caused a 2.7-fold sustained increase in in Cai2+ concentration ([Ca2+]i) and a ninefold sustained increase in pepsinogen secretion. With [Ca2+]o = 0, TG caused a transient 66% increase in [Ca2+]i and a 50% increase in pepsinogen secretion. The time course of TG-induced pepsinogen secretion correlated with the time course of TG-induced increases in [Ca2+]i. These data demonstrated that Ca2+ influx itself was a potent stimulant of pepsinogen secretion. We further focused on the roles of increasing [Ca2+]i from Cai2+ stores. With or without extracellular Ca2+ (Cao2+) present, addition of CCK-8 (0.1 microM) 10 min after TG caused no further increase in [Ca2+]i, demonstrating depletion of the inositol 1,4,5-trisphosphate-sensitive pool. The Ca(2+)-mobilizing agent CCK-8 caused no pepsinogen secretion 10 min after TG preincubation, demonstrating that mobilization of Ca2+ from intracellular stores was important in the rapid initial phase stimulation of pepsinogen secretion caused by CCK-8. In contrast, preincubation with TG had no effect on pepsinogen secretion by secretin, an agent that increases adenosine 3',5'-cyclic monophosphate. A 6-min preincubation with TG potentiated the subsequent stimulation of pepsinogen secretion caused by secretin in the presence of Cao2+ where [Ca2+]i remained elevated. However, TG-induced potentiations of secretin-stimulated pepsinogen secretion was abolished once [Ca2+]i had returned to the basal level in the absence of Cao2+.(ABSTRACT TRUNCATED AT 400 WORDS)
Assuntos
Cálcio/fisiologia , Mucosa Gástrica/metabolismo , Pepsinogênios/metabolismo , Terpenos/farmacologia , Animais , Relação Dose-Resposta a Droga , Sinergismo Farmacológico , Mucosa Gástrica/citologia , Cobaias , Cinética , Masculino , Extratos Vegetais/farmacologia , Secretina/farmacologia , Sincalida/farmacologia , TapsigarginaRESUMO
The possibility that vasoactive intestinal polypeptide (VIP) may facilitate the nicotine-mediated induction of adrenal medullary tyrosine hydroxylase (TH) was investigated with primary cultures (5-7 days in vitro) of bovine adrenal chromaffin (BAC) cells. Exposure of BAC cells to 100 microM nicotine led to only a marginal increase in the amount of TH mRNA, TH protein, and TH activity. VIP, alone or in the presence of a phosphodiesterase inhibitor, produced a marked increase in TH mRNA, TH protein, and TH activity. Moreover, VIP together with nicotine, at concentrations that alone were devoid of effect, increased the amount of TH mRNA and TH activity. A synergistic effect of VIP and nicotine on cAMP accumulation in BAC cells was also apparent. The marginal effects of large doses of nicotine on both cAMP accumulation and TH induction were blocked completely by hexamethonium but were also partially inhibited by the VIP antagonist [p-chloro-D-Phe6,Leu17]-VIP. Nicotine may, therefore, stimulate the release of VIP from cultured BAC cells and VIP, in turn, by increasing cAMP, may synergize with nicotine to enhance TH gene expression.
Assuntos
Medula Suprarrenal/enzimologia , Grânulos Cromafim/enzimologia , Receptores Colinérgicos/efeitos dos fármacos , Tirosina 3-Mono-Oxigenase/biossíntese , Peptídeo Intestinal Vasoativo/farmacologia , Medula Suprarrenal/efeitos dos fármacos , Animais , Northern Blotting , Bovinos , AMP Cíclico/metabolismo , Eletroforese em Gel de Ágar , Indução Enzimática , Bloqueadores Ganglionares/farmacologia , Hexametônio , Compostos de Hexametônio/farmacologia , Nicotina/farmacologia , RNA Mensageiro/metabolismo , Sistemas do Segundo Mensageiro , Secretina/farmacologia , Tirosina 3-Mono-Oxigenase/genéticaRESUMO
MK-329 (formerly L-364,718) is a new nonpeptide antagonist for the peripheral (type-A) cholecystokinin (CCK) receptor, which has proved effective in blocking the actions of both exogenous and endogenous CCK in several species. To evaluate the effect of MK-329 on CCK-stimulated pancreaticobiliary output in man, six normal subjects received 10 mg MK-329 or placebo orally in a randomized, crossover fashion, before a background intravenous infusion of secretin (5 pmol/kg/h) and two doses of CCK-8 (approximately 15 and 40 pmol/kg/h, each for 1 h). Gastric and duodenal juice were aspirated separately via two double-lumen tubes, with 51Cr-ethylene-diaminetetraacetic acid as a duodenal marker. After placebo treatment the background infusion of secretin produced maximum plasma concentrations of secretin similar to postprandial values, averaging about 5 pM. After placebo treatment the low dose CCK-8 infusion (15 pmol/kg/h) increased circulating CCK concentrations from basal levels of 1.8 +/- 0.2 pM to levels similar to those observed postprandially, averaging 9.2 +/- 1.3 pM, and the high dose of CCK-8 (40 pmol/kg/h) induced supraphysiologic levels of CCK, averaging 23.4 +/- 3.2 pM. Plasma concentrations of secretin and CCK were not significantly different during MK-329 treatment. As expected, infusion of CCK-8 at both doses stimulated pancreatic exocrine secretion and gallbladder contraction in placebo controls, as indicated by increases in the output of trypsin, amylase, bicarbonate, and bilirubin. Whereas MK-329 did not significantly reduce basal pancreatic secretion, the integrated incremental output of trypsin, amylase, and bicarbonate in response to stimulation with the low (physiologic) CCK dose was inhibited by 74% (p less than 0.01), 89% (NS), and 75% (p less than 0.05), respectively. Basal bilirubin output was virtually abolished after treatment with MK-329, and the response to the low dose of CCK was reduced by 98% (p less than 0.01), indicating almost complete inhibition of gallbladder contraction at physiologic circulating concentrations of CCK. It is concluded that MK-329 is an orally active antagonist of CCK-stimulated pancreaticobiliary output in man and could thus be utilized to explore the physiologic regulation of the exocrine pancreas and gallbladder by CCK.
Assuntos
Benzodiazepinonas/farmacologia , Colecistocinina/antagonistas & inibidores , Vesícula Biliar/efeitos dos fármacos , Pâncreas/efeitos dos fármacos , Adulto , Amilases/análise , Análise de Variância , Bicarbonatos/análise , Bilirrubina/análise , Colecistocinina/sangue , Colecistocinina/farmacologia , Devazepida , Método Duplo-Cego , Duodeno/química , Suco Gástrico/química , Humanos , Secreções Intestinais/química , Masculino , Pâncreas/enzimologia , Pâncreas/metabolismo , Radioimunoensaio , Secretina/sangue , Secretina/farmacologia , Estimulação Química , Tripsina/análiseRESUMO
We investigated the effects of endogenous and exogenous cholecystokinin (CCK) on pancreatic exocrine secretion, in particular that of bicarbonate. In six dogs prepared with gastric cannulas and Thomas duodenal cannulas, intraduodenal administration of corn oil (Lipomul) incubated with hog pancreatic enzymes significantly increased pancreatic secretion of both bicarbonate and protein. Increase in pancreatic secretion of both bicarbonate and protein was accompanied by the increase in plasma CCK concentration. However, the increase in bicarbonate as well as protein secretion was blocked by proglumide, a CCK antagonist, given intravenously. In contrast, intraduodenal infusion of undigested Lipomul failed to stimulate the pancreatic exocrine secretion or release of endogenous CCK. These observations indicate that endogenous CCK plays an important role in secretion of both bicarbonate and protein stimulated by digested corn oil. In a group of four dogs with pancreatic fistulas, intravenous infusion of CCK potentiated the stimulatory effect of secretin on pancreatic bicarbonate secretion. The stimulatory effect as well as potentiating effect of CCK on pancreatic bicarbonate secretion was blocked by infusion of proglumide. We conclude that endogenous CCK plays a significant role in fat-stimulated pancreatic secretion, and it is apparent that both endogenous CCK and secretin are equally important for stimulation of pancreatic bicarbonate secretion, which results from potentiation of the action of the two hormones.
Assuntos
Bicarbonatos/metabolismo , Água Corporal/metabolismo , Colecistocinina/fisiologia , Pâncreas/metabolismo , Animais , Óleo de Milho/farmacologia , Cães , Feminino , Infusões Parenterais , Masculino , Proglumida/farmacologia , Secretina/farmacologia , Sincalida/farmacologiaRESUMO
Effects of a number of possible trophic factors on growth of guinea pig pancreatic duct epithelial monolayers were investigated. Isolated fragments of main and interlobular ducts were prepared and explanted onto both tissue culture plastic and thick gels of type I collagen. Monolayers growing out from explants were first cultured in a basal medium for 3 or 4 days. Next, the medium was supplemented individually with bombesin, carbachol, caerulein, epidermal growth factor (EGF), secretin, 12-O-tetradecanoylphorbol 13-acetate (TPA), or vasoactive intestinal peptide (VIP). Cells were cultured in the absence or presence of these possible trophic factors, and monolayer areas were determined morphometrically at 0, 2, and 4 days. Rate of growth was determined from increase in area over each 2-day period. Monolayers grown in basal medium alone on plastic increased to 479% of initial area over the 4-day test period; those grown on collagen increased to 523%. Explants cultured in presence of bombesin, carbachol, caerulein, secretin, TPA, and VIP on either substrate grew at rates not significantly different from those cultured in basal medium. By contrast, duct monolayers grown on plastic or collagen in presence of 10 nM EGF expanded in area to 722 and 1,070%, respectively, of their initial areas. The EC50 for this trophic effect was approximately 1 nM. These results show that EGF exerts a potent trophic effect on guinea pig pancreatic duct cells in vitro but also indicate that cell division in the pancreatic main and interlobular ducts is not regulated by caerulein and related peptide hormones that have been reported to have growth-promoting effects on exocrine pancreas in vivo.
Assuntos
Ductos Pancreáticos/citologia , Animais , Bombesina/farmacologia , Carbacol/farmacologia , Divisão Celular/efeitos dos fármacos , Células Cultivadas , Ceruletídeo/farmacologia , Fator de Crescimento Epidérmico/farmacologia , Células Epiteliais , Epitélio/efeitos dos fármacos , Cobaias , Cinética , Microscopia de Contraste de Fase , Técnicas de Cultura de Órgãos , Secretina/farmacologia , Acetato de Tetradecanoilforbol/farmacologia , Peptídeo Intestinal Vasoativo/farmacologiaRESUMO
The effects of a copper-deficient diet supplemented with D-penicillamine on both the structure and exocrine function of the pancreas were studied in rabbits. The pancreases of the animals kept on this diet for 13 weeks showed lesions in the acinar tissue but not in ductal or islet tissue. The weights of the glands were markedly reduced. Copper deficiency caused a slight (non-significant) reduction in the flow of pancreatic juice and in sodium output and significantly decreased chloride output (p less than 0.01). No changes were observed in bicarbonate and potassium output. Total protein and amylase secretions were maintained at clearly detectable levels in pancreatic juice, although they decreased significantly (p less than 0.01 and p less than 0.001, respectively), in the copper-deficient animals compared with the controls. The pancreases of the animals kept on a copper-deficient diet were seen to maintain their capacity for response to secretin, cholecystokinin (CCK), and bethanechol. Our results show that rabbit pancreatic acinar tissue is less sensitive to copper deficiency than that of other species studied. Additionally, the data on secretion of water and electrolytes suggest that in the rabbit the acinar cells may contribute to the secretory process with a chloride-rich fluid.
Assuntos
Cobre/deficiência , Dieta/efeitos adversos , Pâncreas/patologia , Amilases/metabolismo , Animais , Compostos de Betanecol/farmacologia , Cloretos/metabolismo , Colecistocinina/farmacologia , Cobre/farmacologia , Masculino , Pâncreas/metabolismo , Pâncreas/fisiopatologia , Suco Pancreático/metabolismo , Penicilamina/farmacologia , Proteínas/metabolismo , Coelhos , Secretina/farmacologia , Sódio/metabolismo , Equilíbrio Hidroeletrolítico/efeitos dos fármacosRESUMO
If gastric emptying plays a significant role in the satiety produced by exogenous cholecystokinin octapeptide (CCK-8) then 1) the effects on emptying and feeding should share similar kinetics and 2) peptides that inhibit emptying should also inhibit feeding. In the first experiment, CCK-8 (5.6 micrograms/kg) injected immediately before the introduction of an intragastric load (10 ml saline) or presentation of a test meal (15% sucrose) produced a rapid inhibition of both emptying and feeding. In contrast, the same dose administered 15 min before testing caused no inhibition of emptying, even though it retained the ability to reduce meal size. In experiment 2, the abilities of the peptides pentagastrin (100 micrograms/kg), bombesin (8 and 16 micrograms/kg), and secretin (2.86, 14.3, and 28.6 micrograms/kg) to reduce food intake and inhibit emptying were tested. Pentagastrin influenced neither food intake nor gastric emptying. Bombesin caused a small transient delay in emptying but a large and sustained eating suppression. However, a high dose of secretin caused no significant reduction of food intake, in spite of the fact that it inhibited emptying to the same degree as 1.4 micrograms/kg CCK-8, which does reduce intake. These results suggest that the inhibition of emptying by CCK is not sufficient to explain the satiety effect of CCK-8.