Glucagon-37 (oxyntomodulin) and glucagon-29 (pancreatic glucagon) in human bowel: analysis by HPLC and radioreceptorassay

A method for assaying specifically the biologically active peptides Glucagon-37 (G-37/Oxyntomodulin/bioactive Enteroglucagon) and Glucagon-29 (G-29/pancreatic Glucagon) has been developed by use of high performance liquid chromatography (HPLC) of crude tissue extracts followed by radioreceptorassay in liver membranes. The peaks observed with this method in samples from human bowel have also been analysed in two other assays: stimulation of cyclic AMP accumulation in gastric glands and radioimmunoassay. Owing to the different patterns of activity of porcine G-37 and G-29 in these assays, the comparison of the data obtained allows to discriminate between the two peptides. The same behaviour in both HPLC and the three assays of the human peaks on one hand and the porcine peptides on the other strongly suggests that human intestine contains a very similar or the same molecules as that isolated from the porcine tissues. Whatever the portion of small intestine, G-37 represented ca 90% of G-37 + G-29. A decreasing concentration gradient of both G-37 and G-29 was also observed from ileum to descending colon.     

Effect of glucagon on the secretory process in the rat exocrine pancreas

Summary Glucagon was infused into conscious rats in doses of 10 to 80 g/h for periods up to 24 h. The effect on the secretory process of the exocrine pancreas was studied in vitro using isolated pancreatic lobules. A pronounced inhibition of the rate of protein synthesis and discharge of stored and newly synthesized proteins combined with increased enzyme content in the pancreas were observed after 30 min infusion. This effect was absent after longer infusion periods of up to six hours. After 12 to 24 h infusions a marked degranulation and decrease in enzyme content was observed. While the rate of protein synthesis was not significantly enhanced, both the basal and stimulated discharge of enzymes from the pancreas were increased. The results suggest a biphasic response of the pancreas to prolonged glucagon infusion.Dedicated to Professor Helmut Ferner, Vienna, Austria, on the occasion of his 65th birthday     

Activity of oxyntomodulin on gastric acid secretion induced by histamine or a meal in the rat

Conscious rats with chronic gastric fistula were trained for drinking a 14-ml milk meal. The activity of an intestinal hormone, oxyntomodulin (OXM), was studied in this model and compared to that observed when histamine was the stimulus. Under histamine (0.25 mg·kg⁻¹·h⁻¹) stimulation, OXM at doses (60–120 pmol·kg⁻¹·h⁻¹) that induced physiological circulating levels inhibited gastric acid secretion up to 50%. Under meal stimulation, OXM reduced up to 29% acid secretion at doses (1–1.5 nmol·kg⁻¹·h⁻¹) inducing supraphysiological levels. We conclude that at physiological concentrations OXM cannot counteract the complex processes triggered by a meal. OXM would be a component of enterogastrone, a combination of several intestinal hormones acting in synergy. The OXM action is related to pathways recognizing the C-terminal 19–37 moiety of the molecule.     

Inhibitory effect of pancreastatin on pancreatic exocrine secretion in the conscious rat

The effect of newly discovered pancreastatin on pancreatic secretion stimulated by a diversion of bile-pancreatic juice (BPJ) from the intestine was examined in the conscious rat. Exogenous pancreastatin infusion (20, 100 and 200 pmol/kg.h) inhibited pancreatic protein and fluid outputs during BPJ diversion in a dose-dependent manner. Pancreastatin did not affect plasma cholecystokinin (CCK) concentrations. Pancreastatin (100 pmol/kg.h) inhibited CCK-stimulated pancreatic secretion, but did not inhibit secretin-stimulated pancreatic secretion. Pancreastatin alone, however, did not affect basal pancreatic secretion. In contrast, pancreastatin (10(-10)-10(-7)M) did not suppress CCK-stimulated amylase release from isolated rat pancreatic acini. These results indicate that pancreastatin has an inhibitory action on exocrine function of the pancreas. This action may not be mediated by direct mechanisms and nor via an inhibition of CCK release. It is suggested that pancreastatin may play a role in the regulation of the intestinal phase of exocrine pancreatic secretion.     

Effect of colonic distension on exocrine pancreatic secretion of the conscious rat

In the conscious rat the increasing distension of the colon (from 2 to 6 ml) induced a slight decrease of exocrine pancreatic secretion which was more important for protein output than for volume. Nevertheless this inhibitory effect was never significant even with the important distension of 6 ml which induced a visible uneasiness in the animal.     

Three-day continuous drainage of pancreatic juice in the cortisone-treated conscious rat: Analysis of enzyme activities and ultrastructural changes

Summary We have investigated the short-term effects of hydrocortisone (60 mg/kg per day) and placebo on basal and stimulated pancreatic secretion in the conscious rat. Volume and enzyme secretion were determined; fine structural changes were examined simultaneously.The pancreatic and bile ducts were cannulated separately; pancreatic juice was drained via an isolated fistula, and bile was recirculated into the duodenum. The application of hydrocortisone led to an almost complete inhibition of the secretory response of the exocrine pancreas when stimulated with 0.25 U secretin in combination with 5 × 10^-8 g caerulein per h. It strongly affected the secretion rates of volume, protein, lipase, chymotrypsin, trypsin and carboxypeptidase, whereas the secretion rate of alpha-amylase continued to show a slight increase after stimulation.After stimulation with secretin and caerulein, the hydrocortisone-treated animals showed a higher density of zymogen granules in the acinar cell and an increase in the number of autophagic vacuoles in comparison to the equally stimulated placebo-treated rats.It is concluded that the short-term inhibition of pancreatic secretion by hydrocortisone occurs largely as a result of an inhibition of cellular enzyme discharge.Supported by the Deutsche Forschungsgemeinschaft, Ga 279     

The effect of oleate on pancreatic and bile secretion in the conscious rat

The effects of sodium oleate infused into either the duodenum or the terminal ileum on bile and pancreatic secretion were examined in the conscious rat. Rats were prepared with cannulae draining pure bile and pancreatic juice separately, and with an ileal and two duodenal cannulae. A 40 mM taurocholate solution containing 7 mg/ml bovine trypsin was infused into the duodenum throughout the experiment to replace diverted bile-pancreatic juice to maintain the normal regulation of pancreatic secretion. The intraduodenal infusion of sodium oleate significantly increased pancreatic juice flow, protein, and bicarbonate outputs, whereas it did not affect bile secretion. Intravenous infusion of proglumide (300 mg/kg/hr) did not inhibit pancreatic secretion stimulated by intraduodenal infusion of sodium oleate. An intravenous infusion of atropine (100 micrograms/kg/hr) attenuated protein and fluid secretions but not that of bicarbonate in response to intraduodenal oleate. In contrast, the intraileal infusion of oleate had no effect on pancreatic secretion, whereas it decreased bile flow, bicarbonate, and bile salt outputs. In conclusion, sodium oleate introduced in the duodenum stimulates pancreatic secretion but oleate in the terminal ileum inhibits bile secretion.     

Effect of pancreatic polypeptide-antiserum on bombesin-stimulated pancreatic exocrine secretion in dogs

Bombesin is a potent stimulus of both pancreatic protein secretion and plasma pancreatic polypeptide (PP) release in dogs. Physiological plasma levels of PP have been shown to inhibit pancreatic exocrine secretion in dogs. We examined the question whether the concomitant release of PP exerts a suppressive action on the pancreatic exocrine response to bombesin in dogs by measuring pancreatic exocrine secretion with and without in vivo immunoneutralization of PP with a high affinity PP-antiserum. Bombesin was infused in a dose of 150 ng/kg·hr, resulting in a rise of plasma PP from 24±5 to 224±25 pM (p<0.01). Before this bombesin infusion, 7 ml of normal rabbit serum had been administered to the dogs (n=8). At a later stage, the study was repeated after administration of 7 ml of PP-antiserum to the same animals. The bombesin induced increase in pancreatic exocrine secretion during administration of PP-antiserum (flow rate 24±10 ml/hr, protein output 1.35±0.43 g/hr, and bicarbonate output 3.25±1.42 mmol/hr) was not significantly different from that during control rabbit serum (flow rate 21±7 ml/hr, protein output 1.26±0.38 g/hr, and bicarbonate output 3.18±1.10 mmol/hr). It is therefore concluded that the pancreatic exocrine response to bombesin is not affected by the concomitant secretion of PP.     

Thyrotropin-releasing hormone injected intracerebroventricularly in the rat stimulates exocrine pancreatic secretion via the vagus nerve

The effects of intracerebroventricular (i.c.v.) injection of synthetic thyrotropin-releasing hormone (TRH) and its analogue (gamma-butyrolactone-gamma-carbonyl-His-Pro-NH2) were tested in anesthetized rats fitted with pancreatic cannula. TRH injection induced dose-related increases in flow of pancreatic juice, protein output, and amylase output, each reaching a maximum within 10 min. Higher doses of TRH induced longer responses. Injection of the TRH analogue also caused dose-related secretory responses of the exocrine pancreas. The dose-related secretory responses to TRH and the TRH analogue were similar except that the responses to the highest dose of TRH analogue (1600 pmol/100 g b.w.) were significantly higher. Intravenous injection of TRH and the TRH analogue induced little, if any, secretory response of the exocrine pancreas. The effects of i.c.v. injection of TRH and the TRH analogue were completely abolished after bilateral subdiaphragmatic vagotomy. In addition to the secretory effects on the exocrine pancreas, i.c.v. injection of TRH and the analogue caused hyperglycemia, tachycardia, and tear secretion, but the intravenous injection of these peptides had no effect.     

Nitric oxide and the pancreas: Morphological base and role in the control of the exocrine pancreatic secretion

The distribution of nitric oxide synthase in both neuronal and non-neuronal pancreatic tissues and the role of nitric oxide in the control of exocrine pancreatic secretion are reviewed in this article. Earlier reports based on in vivo studies suggested that nitric oxide can affect the secretory activity of the exocrine pancreas through changes in pancreatic blood flow. More recently, the employment of either nitric oxide synthase inhibitors or nitric oxide donors in in vitro preparations has provided evidence that nitric oxide can exert a direct action on this gland independently on its vascular effects. Most research in this area seems to indicate that modulation of exocrine pancreatic function by nitric oxide is exerted via activation of guanylate cyclase and generation of cGMP, although other pathways cannot be excluded. Experiments performed over the last year in our laboratory reveal a novel and interesting mechanism based on the ability of nitric oxide to control the release of endogenous neurotransmitter in the pancreas and, subsequently, the nerve-mediated enzyme secretion.     

Effect of proglumide, a cholecystokinin receptor antagonist, on caerulein-stimulated pancreatic enzyme secretion and pancreatic polypeptide release in the dog

In five conscious dogs we studied the effect of proglumide, a cholecystokinin (CCK) antagonist, on caerulein-stimulated pancreatic secretion and release of pancreatic polypeptide (PP). Graded doses of caerulein (15-240 ng/kg per h) were infused intravenously. Experiments were repeated with a fixed infusion of proglumide (40 mg/kg per h). Release of PP following increasing doses of caerulein was significantly inhibited by proglumide (P less than 0.01). However, proglumide did not significantly affect caerulein-stimulated pancreatic protein secretion. Proglumide might be useful in defining the physiological role of CCK.     

The role of aquaporin water channels in fluid secretion by the exocrine pancreas

The mammalian exocrine pancreas secretes a near-isosmotic fluid over a wide osmolarity range. The role of aquaporin (AQP) water channels in this process is now becoming clearer. AQP8 water channels, which were initially cloned from rat pancreas, are expressed at the apical membrane of pancreatic acinar cells and contribute to their osmotic permeability. However, the acinar cells secrete relatively little fluid and there is no obvious defect in pancreatic function in AQP8 knockout mice. Most of the fluid secreted by the pancreas is generated by ductal epithelial cells, which comprise only a small fraction of the gland mass. In the human pancreas, secretion occurs mainly in the intercalated ducts, where the epithelial cells express abundant AQP1 and AQP5 at the apical membrane and AQP1 alone at the basolateral membrane. In the rat and mouse, fluid secretion occurs mainly in the interlobular ducts where AQP1 and AQP5 are again co-localized at the apical membrane but appear to be expressed at relatively low levels. Nonetheless, the transepithelial osmotic permeability of rat interlobular ducts is sufficient to support near-isosmotic fluid secretion at observed rates. Furthermore, apical, but not basolateral, application of Hg²⁺ significantly reduces the transepithelial osmotic permeability, suggesting that apical AQP1 and AQP5 may contribute significantly to fluid secretion. The apparently normal fluid output of the pancreas in AQP1 knockout mice may reflect the presence of AQP5 at the apical membrane.     

Effect of insulin on exocrine pancreatic secretion in healthy and diabetic anaesthetised rats

This investigation characterised the effects of exogenous insulin on exocrine pancreatic secretion in anaesthetised healthy and diabetic rats. Animals were rendered diabetic by a single injection of streptozotocin (STZ, 60 mg kg^–1 I.P.). Age-matched controls were injected citrate buffer. Rats were tested for hyperglycaemia 4 days after STZ injection and 7–8 weeks later when they were used for the experiments. Following anaesthesia (1 g kg^–1 urethane I.P.), laparotomy was performed and the pancreatic duct cannulated for collection of pure pancreatic juice. Basal pancreatic juice flow rate in diabetic rats was significantly (p < 0.001) increased whereas protein and amylase outputs were significantly (p < 0.001) decreased compared to control rats. Insulin (1 IU, I.P.) produced in healthy rats significant increases in pancreatic flow rate, amylase secretion and protein output compared to basal (p < 0.05). Insulin action also included a reduction in blood glucose (152.7 ± 16.9 mg dl^–1, n= 6, prior to insulin and 42.0 ± 8.4 mg dl^–1, n= 4, 100 min later). In fact, flow rate and glycaemia showed a strong negative correlation (p < 0.01, Pearson). Pretreatment with atropine (0.2 mg kg^–1, I.V.) abolished the effects of insulin on secretory parameters despite a similar reduction in glycaemia; in this series of experiments the correlation between flow rate and blood glucose was lost. In diabetic rats, insulin (4 IU, I.P.) did not modify exocrine pancreatic secretion. There was a fall in blood glucose (467.6 ± 14.0 mg dl^–1, n= 10, prior to insulin and 386.6 ± 43.6 mg dl^–1, n= 7, 120 min later). Rats, however, did not become hypoglycaemic. Similar results were observed in diabetic atropinized rats. The results of this study indicate that the effects of insulin on exocrine pancreatic secretion in anaesthetised healthy rats are mediated by hypoglycaemia-evoked vagal cholinergic activation. (Mol Cell Biochem 261: 105–110, 2004)     

半胱胺对鹅胰液分泌及胰酶活性的影响

目的 :研究半胱胺对鹅胰液分泌及胰酶活性的影响。方法 :12只装有胰腺～十二指肠长久性瘘管的成年鹅。试验采用自身对照 ,试验期在日粮中一次性添加半胱胺 (10 0mg/kgbw)。连续收集计量胰液并测定胰酶活性。 结果 :①半胱胺使鹅胰液的分泌速率较对照期显著上升 (2 4 0 .16 % ,P <0 .0 1) ,其中白天升高 2 34.4 5 % ,夜间增高了2 5 3.70 %。②试验期单位容积胰蛋白酶的活性较对照期升高了 4 9.0 5 % (P <0 .0 1) ,而胰脂肪酶和胰淀粉酶的活性却较对照期分别降低了 2 5 .4 4 %和 2 1.95 % ,且变化幅度具有昼夜的差别。③试验期胰腺每小时分泌胰蛋白酶、胰脂肪酶和胰淀粉酶的总活性较对照期分别升高 4 0 6 .88% (P <0 .0 1)、15 3.5 8% (P <0 .0 1)和 16 6 .5 9% (P<0 .0 1) ,白天增加的幅度较夜晚大。结论 :半胱胺能促进鹅胰液的分泌 ,增加胰液中蛋白酶、脂肪酶和淀粉酶分泌的总量 ,从而提高鹅对饲料的消化能力 ,适应机体生长对营养的需求     

Pituitary adenylate cyclase-activating polypeptide (PACAP): occurrence in rodent pancreas and effects on insulin and glucagon secretion in the mouse

Summary Pituitary adenylate cyclase-activating polypeptide (PACAP) is a neuropeptide that occurs in several tissues, e.g., in the gut. We have studied PACAP-like immunoreactivity in the pancreas of rat and mouse, and the effects of PACAP-38 on basal and stimulated insulin and glucagon secretion in the mouse. Immunofluorescence staining demonstrated the presence of PACAP-like immunoreactivity in nerve fibers in both the rat and mouse pancreas. The nerve fibers were seen in the exocrine pancreas and surrounding the islets. Occasionally, the nerve fibers occurred within the islets. Most PACAP-positive nerve fibers innervated the intrapancreatic ganglia, although no nerve cell bodies contained PACAP-like immunoreactivity. In-vivo experiments in mice revealed that basal plasma glucagon levels were increased by PACAP-39 injected intravenously at dose levels exceeding 1.8 nmol/kg. Furthermore, PACAP-38 (7 nmol/kg) potentiated the plasma glucagon response to the cholinergic agonist carbachol (0.16 mol/kg). This potentiation was reduced to simple addition by pretreatment with a combined - and -adrenergic blockade by phentolamine (35 mol/kg) and propranolol (8.5 mol/kg). Moreover, PACAP-38 inhibited a carbachol-induced increase in the level of plasma insulin in the absence but not in the presence of adrenergic blockade. PACAP-38 increased basal plasma insulin levels and increased basal plasma glucose levels 6 min and 10 min, respectively, after injection of the peptide. We conclude that PACAP-like immunoreactivity exists in nerve fibers innervating the mouse and rat pancreas, particularly the intrapancreatic ganglia, and that PACAP-38 augments both basal and carbachol-stimulated glucagon secretion in the mouse.     

The effect of hypothermia on exocrine pancreatic secretion in rabbits

The effects of experimentally induced hypothermia on exocrine pancreatic secretion in rabbits were investigated. During hypothermia the flow of pancreatic juice decreased to 50% of basal values and recovered after rewarming. Hypothermia scarcely affected HCO-3, Cl- and Na+ concentrations but did cause significant alterations in K+ concentrations. During hypothermia and later normothermia a parallel secretion in the enzymes amylase, chymotrypsin and trypsin was seen to take place. Enzyme secretion decreased throughout the experimental period in the rabbits undergoing hypothermia and later normothermia, as in the case of the control animals.     

Freeze-fracture study on the whorls of rough endoplasmic reticulum in the exocrine pancreatic cells of the Japanese newt and African clawed toad

Summary The whorls of rough endoplasmic reticulum (rER) in the exocrine pancreatic cells of starved newts and clawed toads were examined by a freeze-fracture technique. The whorl appeared to be roughly ovoidal in shape and composed of tightly packed, narrow cisternae arranged like the layers of an onion. The clusters of interdigitating projections of the cisternal membranes were located at several places on the whorl. Some of these projections extended to the vesicular rER around the whorl. The fenestra-like, raised or hollowed craters were seldom seen on the fractured membrane faces of the whorls in the exocrine pancreatic cells of the starved newts.     

The influence of potato fibre on exocrine pancreatic secretions and on plasma levels of insulin,secretin and cholecystokinin in growing pigs

The effect of a potato fibre preparation on exocrine pancreatic secretions and on gastrointestinal hormone levels in plasma was studied in three 8 weeks old piglets that were surgically fitted with a jugular vein catheter for blood sampling, a pancreatic duct catheter and a T‐shaped duodenal cannula for collection of pancreatic juice. The animals were fed for 2 weeks a control diet (experimental period 1), thereafter for 2 weeks the control diet supplemented with 2% potato fibre (experimental period 2) and for another 2 weeks the control diet again (experimental period 3). Additionally, intraduodenal (i.d.) infusions of the experimental diet, the control diet and potato fibre as well as i.v. infusions of a solution containing cholecystokinin (CCK) and secretin were administered.

Potato fibre in the diet evoked in tendency an increase in the volume of secretion of pancreatic juice and a significant increase both in the mean values of the total protein content and total activities of lipase, trypsin and a‐amylase when compared to the control diet. The i.d. infusion of the control diet, experimental diet and fibre infusate as well as the i.V. administration of the hormone infusate led to a spontaneous secretory response of the exocrine pancreas. Besides gastrointestinal hormones, such as CCK, other factors such as short chain fatty acids may be involved in the regulation of the exocrine pancreas.     

Effects of pancreatic polypeptide on the pancreatic exocrine secretion stimulated by secretin and cholecystokinin in the conscious pig

Fourteen castrated male Large White pigs, weighing 42.5 +/- 1.0 kg, were fitted with pancreatic and duodenal fistulae for pancreatic secretion studies. Moreover, catheters were placed in a carotid artery for blood sampling and in a jugular vein for peptide infusion. Pancreatic juice was automatically restituted to the animals and continuously sampled for analysis on experimental days. Following an 8-day recovery period, perfusion studies were performed after an overnight fast. After a 30-min basal period, sustained pancreatic flow and protein output were obtained and maintained throughout the assay with secretin (36 pmol/kg/h) and CCK-8 (600 pmol/kg/h) infusion. Then, 200, 400, 600, 800 or 1200 pmol/kg/h of porcine pancreatic polypeptide (PP) were infused for 60 min. Secretin + CCK infusion was continued for 1 h after PP infusion was stopped. Each dose of PP was given on a separate day. Neither pancreatic flow nor bicarbonate output were affected whatever the dose of infused PP. On the contrary, protein concentration and output decreased with the lowest dose of PP (200 pmol/kg/h) and the diminution was more pronounced with the other doses. With 600 pmol/kg/h as well as with 800 and 1200 pmol/kg/h of PP, pancreatic protein output fell to about 20% of values obtained with secretin + CCK. Plasma levels of PP were below or similar to postprandial values for 200, 400 and 600 pmol/kg/h and they were significantly larger with 800 and 1200 pmol/kg/h. Protein concentration and output returned to values obtained with secretin + CCK infusion after cessation of PP infusion. In conclusion, porcine PP given in physiological doses to the pig decreases pancreatic protein output whereas pancreatic flow remains unaffected.     

Atropine depresses release of neurotensin and its effect on the exocrine pancreas

In this study the effect of 10 and 20 μg · kg^?1 · h^?1 atropine sulfate on release and pancreatic effects of neurotensin was studied in 4 dogs. Neurotensin plasma levels rose significantly when a liquid fat preparation was infused intraduodenally. This rise was almost completely abolished by simultaneous infusion of atropine. Atropine further suppressed basal and fat-stimulated output of pancreatic volume, protein, and bicarbonate; it also reduced pancreatic secretion stimulated by an intravenous infusion of low doses (2.5 to 20 pmol · kg^?1 · min^?1) neurotensin. The effect of higher doses (80 and 240 pmol · kg^?1 · min^?1) of neurotensin was less affected.As neurotensin plasma levels in contrast to normal oral feeding did not rise after sham feeding, our findings suggest that release and action of neurotensin may at least in part be dependent on a cholinergic, non-cephalic mechanism.