Effect of vasoactive intestinal polypeptide and somatostatin on secretion of epidermal growth factor and bicarbonate from Brunner''s glands

The effect of VIP and somatostatin on secretion of epidermal growth factor and bicarbonate from Brunner's glands was investigated in the rat. Vasoactive intestinal polypeptide infused in doses of 10 and 100 ng/kg/h significantly increased epidermal growth factor and bicarbonate output, but the concentrations did not change. Somatostatin infused at doses of 1, 10, 100 and 1000 ng/kg/h against a background of VIP 100 ng/kg/h inhibited in dose-dependent fashion the stimulated epidermal growth factor and bicarbonate outputs from rat Brunner's gland pouches. Also basal secretion was inhibited by somatostatin. Infusion of antisomatostatin serum stimulated Brunner's gland secretion. By immunohistochemical studies of rat duodena, it was found that epidermal growth factor, is almost exclusively present in the secretory cells of Brunner's glands. It is concluded that VIP stimulates secretion of epidermal growth factor and bicarbonate from Brunner's glands, an effect which is inhibited by somatostatin. A possible role for somatostatin in the control of Brunner's gland secretion is suggested.     

Adrenergic effects on secretion of epidermal growth factor from Brunner's glands.

The influence of the sympathetic nervous system and adrenergic agonists on flow rate and secretion of epidermal growth factor (EGF) from Brunner's glands has been investigated in the rat. Chemical sympathectomy by administration of 6-hydroxydopamine increased volume secretion and output of EGF from Brunner's glands but depleted the glands of EGF. Infusion of noradrenaline, an alpha-adrenergic agonist, inhibited basal and vasoactive intestinal polypeptide (VIP) stimulated flow rate and output of EGF from Brunner's glands and increased the amount of EGF in the tissue. Vasoactive intestinal polypeptide also increased the amount of EGF in Brunner's gland tissue and this was unchanged after simultaneous infusion of VIP and noradrenaline as well as VIP and isoproterenol, a beta-adrenergic agonist. Isoproterenol had no effect on basal and VIP stimulated secretion of EGF from Brunner's glands. The presence of PAS-positive mucus in Brunner's glands was unchanged during infusion of noradrenaline whereas VIP induced a depletion of Brunner's gland mucus which in turn was prevented by simultaneous infusion of noradrenaline. This study indicates that the sympathetic nervous system influence the volume secretion, output of EGF and mucus content in Brunner's glands probably by activation of alpha-adrenergic pathways.     

Effect of secretin and glucagon on Brunner''s gland secretion in the rat

Brunner's gland secretion in response to infusion of secretin and glucagon was studied in the rat. Secretin was infused in doses of 15, 150 and 1500 ng/kg/h. All dose significantly increased bicarbonate and protein output and depleted Brunner's glands of PAS-positive mucin. Bicarbonate secretion was related to plasma secretin concentration, and a marked stimulatory effect of secretin was found in very low, probably physiological, plasma concentrations. Maximal bicarbonate output was obtained at a plasma concentration of secretin about 20 pmol/l. Glucagon was infused at a rate of 1.0 micrograms/kg/h and did not influence secretion rate or cell morphology. Also large doses of 5.0 and 50.0 micrograms/kg/h had no effect on Brunner's gland secretion. It is concluded that secretin in very low plasma concentrations stimulates secretion of bicarbonate, protein and mucus from Brunner's glands in the rat, while glucagon has no effect, and it is suggested that secretin may be involved in the physiological regulation of Brunner's gland secretion.     

Effects of somatostatin on acute canine experimental pancreatitits

Summary Somatostatin is an inhibitory hormone that decreases the secretion and end organ response of cholecystokinin (CCK). Inhibition of hormonal stimulation of pancreatic exocrine secretion by somatostatin may improve the course of acute pancreatitis. Anesthetized dogs underwent cholecystectomy and cannulation of the pancreatic duct, thoracic, duct, and portal vein. Twenty experiments were performed in random order with 5 dogs in each group. Hourly measurements of lymph flow and portal and thoracic duct amylase were made. Portal blood insulin, glucagon, and CCK concentrations were determined by radioimmunoassay on samples obtained at the beginning and end of the experiments. Pancreatitis was induced by injecting, under, constant pressure, 10 ml bile into the pancreatic duct during 1 min. Somatostatin was administered intravenously (20 μg/kg/hr). After 5 h, the dogs were killed, pancreas glands removed and weighted and tissue samples obtained for histologic evaluation. There was a significant-increase in lymph amylase output and portal venous amylase and CCK concentrations in the dogs with pancreatitis compared to the control dogs. In dogs with pancreatitis, lymphatic amylase secretion and portal CCK concentrations were significantly decreased by somatostatin. Somatostatin did not significantly alter portal amylase concentrations, pancreas gland weights or histologic inflammation when compared to values from dogs with pancreatitis not treated with somatostatin. In bile-induced acute experimental pancreatitis, somatostatin decreases thoracic duct amylase output and may have a protective influence on the course of pancreatitis.     

Effect of intravenous infusion of somatostatin on gastric, pancreatic and biliary secretion in the rat.

The effect of somatostatin (GH-RIH) on gastric, pancreatic and biliary secretion was examined in the anesthetized rats. Intravenous infusion of the hormone, in graded doses ranging from 1 to 16 microgram/kg/hr, produced a dose dependent inhibition of pentagastrin, 1.5 microgram/kg/hr, induced acid secretion, reaching about 54% of control level at the dose of 16 microgram/kg/hr. Somatostatin, 4, 16 and 64 microgram/kg/hr, inhibited the pancreatic juice volume stimulated by intravenous injection of secretin, 1 u/kg, as a bolus. Somatostastin, 2, 8, 32 and 128 microgram/kg/30 min, did not change the bile flow rate in control period. Somatostatin may play a physiological role in the regulation of the secretory process of the stomach and pancreas.     

Effect of somatostatin 14 on pure human pancreatic secretion

While it is well known that large doses of somatostatin inhibit human pancreatic enzyme secretion, it is still unknown whether low doses are also effective and whether the peptide is able to inhibit bicarbonate production. Eight subjects with external transduodenal drainage of the main pancreatic duct performed after biliary tract surgery were studied. Somatostatin was infused at progressively increasing rates of 0.05, 0.15, 0.45, and 1.35 g/kg/hr, for 30 min/dose, during pancreatic stimulation with secretin, 25 ng/kg/hr, and cerulein, 10 ng/kg/hr. Somatostatin, at the dose of 0.05 g/kg/hr (shown to produce blood levels similar to those measured after a meal) did not affect pancreatic secretion in any of the subjects. The successive three higher doses caused a significant and dose-dependent inhibition of protein concentration and output and of bicarbonate output. Bicarbonate concentration was slightly but significantly reduced only by the two highest doses of somatostatin. At each dose level, the inhibition of protein output was much more marked than the inhibition of bicarbonate output. The maximal inhibition of protein output (at 1.35 g/kg/hr somatostatin) was 73.9±5.4%, and that of bicarbonate output was 55.9±6.4%. The results demonstrate that: (1) the administration of somatostatin at a low dose level does not affect human exocrine pancreatic secretion, at least under the experimental conditions of this study; and (2) the administration of larger doses of somatostatin inhibits pancreatic secretion of both protein and bicarbonate dose-dependently. The inhibitory effect on protein output is significantly greater than that on water and bicarbonate production.     

Effects of somatostatin on acute canine experimental pancreatitis

Somatostatin is an inhibitory hormone that decreases the secretion and end organ response of cholecystokinin (CCK). Inhibition of hormonal stimulation of pancreatic exocrine secretion by somatostatin may improve the course of acute pancreatitis. Anesthetized dogs underwent cholecystectomy and cannulation of the pancreatic duct, thoracic duct, and portal vein. Twenty experiments were performed in random order with 5 dogs in each group. Hourly measurements of lymph flow and portal and thoracic duct amylase were made. Portal blood insulin, glucagon, and CCK concentrations were determined by radioimmunoassay on samples obtained at the beginning and end of the experiments. Pancreatitis was induced by injecting, under constant pressure, 10 ml bile into the pancreatic duct during 1 min. Somatostatin was administered intravenously (20 micrograms/kg/hr). After 5 h, the dogs were killed, pancreas glands removed and weighed and tissue samples obtained for histologic evaluation. There was a significant increase in lymph amylase output and portal venous amylase and CCK concentrations in the dogs with pancreatitis compared to the control dogs. In dogs with pancreatitis, lymphatic amylase secretion and portal CCK concentrations were significantly decreased by somatostatin. Somatostatin did not significantly alter portal amylase concentrations, pancreas gland weights or histologic inflammation when compared to values from dogs with pancreatitis not treated with somatostatin.     

Proportional inhibition of glucose-induced insulin release by somatostatin

To test the hypothesis that somatostatin in a proportional inhibitor of glucose-induced insulin release, we examined the effect of somatostatin on the release of insulin stimulated either by endogenous signals (basal), by glucose infusion, or by glucose injection. Somatostatin infusions (1.7 μg/min × 30 min) produced a decrement of basal insulin output from the right lobe of the canine pancreas that was proportional to the initial rate of basal insulin secretion (r = −0.87, p < 0.001, n = 16) Glucose infusions of 1–6 mg/kg/min produced much higher rates of insulin output; again, the decrement of insulin output produced by somatostatin correlated with the initial rate of insulin secretion (r = −0.68, p < 0.01, n = 16). Rapid intravenous injection of either 2 or 20 g of glucose produced a wide range of acute insulin responses (AIR). Somatostatin produced a decrement that was proportional to the original magnitude of the AIR (r = −0.70, p < 0.005, n = 16). Thus, the absolute amount of inhibition produced by somatostatin increases, not decreases, with the magnitude of the stimulus. These data suggest that the inhibitory effect of somatostatin cannot be overcome by increasing the size of glucose stimulus. Thus, exogenous somatostatin, and presumably endogenous somatostatin as well, will produce an inhibitory effect at any physiologic level of glucose stimulation that the β-cell receives.     

Secretin is an enterogastrone in humans

We studied in humans the effect of exogenous secretin in a physiological dose on gastric acid secretion stimulated by pentagastrin and postprandial plasma gastrin concentration. Two doses of pentagastrin, 80 and 160 pmol/kg/hr were used to stimulate gastric acid secretion. Secretin in two doses, 2.8 and 5.6 pmol/kg/hr were tried to study the response on stimulated gastric acid secretion. Intravenous secretin in a dose of 5.6 pmol/kg/hr significantly inhibited the gastric acid output stimulated by intravenous pentagastrin in a dose of 160 pmol/kg/hr, from 11.25±1.5 to 5.99±1.34 meq/hr, while lower dose of intravenous secretin (2.8 pmol/kg/hr) failed to inhibit the gastric acid output stimulated by the same dose of pentagastrin. However, the lower dose of intravenous secretin (2.8 pmol/kg/hr) inhibited the gastric acid output significantly from 8.78±1.21 to 6.37±1.62 meq/hr when gastric secretion was stimulated by the lower dose of pentagastrin (80 pmol/kg/hr). The plasma concentrations of secretin during intravenous secretin in a dose of 2.8 pmol/kg/hr was similar to postprandial plasma concentrations of secretin as previously reported. Doubling the dose of intravenous secretin resulted in almost twofold higher plasma concentrations than postprandial plasma concentrations. In addition, the low dose of secretin (2.8 pmol/kg/hr) suppressed the integrated postprandial gastrin response from 13.9±3.7 to 11.2±2.8 ng/min/ml (P=0.05) when endogenous release of secretin was blocked by intravenous cimetidine. Since the dose of pentagastrin and secretin employed in this study fell in a physiologic range, the inhibitory effect of secretin on stimulated gastric acid secretion appears to be a physiologic action in humans. Contrary to the findings in dogs, the inhibitory action of secretin on gastrin release was not statistically significant but was highly suggestive.     

Somatostatin inhibits the effect of secretin on bile flow and on hepatic bilirubin transport in the rat.

Increasing amounts of porcine secretion (0.05 to 2.00 clinical units/h/100 g body wt) given to rats during a continuous infusion of bilirubin, increased bile flow and the apparent maximal biliary excretion of bilirubin ('Tm'). This increment was caused by an enhanced biliary output of bilirubin monoconjugates. The effect was dose dependent but maximal at a secretin infusion of 0.80 CU. Somatostatin 0.2 and 0.8 microgram/h/100 g body wt caused a dose related inhibition of the hepatic effects of secretin both on bile flow and on biliary output of bilirubin conjugates. As secretin elicits the release of somatostatin, a feed-back system could be envisaged whereby the somatostatin released stops the effects of secretin.     

Respiratory exchanges and acid-base balance during perfusion ofex vivo isolated pancreas

We used the technique ofex vivo isolated pancreas, perfused with whole heparinized blood. The organ was stimulated by secretin (G.I.H. Stockolm, 0.1–5.0 clinic units/hr), and/or carbamylcholine (100–200 g/hr). Oxygen consumption was increased under stimulation. This increase was a function of the dose of secretin and also of the bicarbonate output in the juice. Oxygen uptake increased further when carbamylcholine was superimposed on secretin. This extra increase was independent of hemodynamic conditions of the organ perfusion. Arteriovenous difference in oxygen saturation did not increase when the gland was stimulated. It tended to decrease when the stimulation resulted in a marked vasodilatation. Thus, oxygen needs seemed to be neither the limiting factor of the response to a given stimulation nor the triggering mechanism of functional vasodilation. Values of pCO₂ were spread over a wide range from one experiment to another. However, it could be observed that CO₂ effux into the vein decreased under stimulation by secretin; in most experiments, CO₂ efflux was even replaced by an apparent consumption of CO₂ during the perfusion of the stimulated gland. Furthermore, arteriovenous pH difference increased following secretin stimulation. This increase was dose-related to secretin. These facts are discussed under the background of theories recently proposed for bicarbonate secretion.This study was supported by Contrat No. 73.3.054.4 from INSERM.     

Distribution and release of epidermal growth factor in man.

Epidermal growth factor (EGF) is localised in man to salivary and Brunner's glands. It is present in large concentrations in saliva and duodenal contents but the mechanisms of its release have been little studied. This study carried out on four groups of healthy subjects was designed to determine the distribution and the release of immunoreactive EGF (IR-EGF) in salivary, gastric, duodenal, and pancreatic secretions. Under basal conditions, the concentrations of IR-EGF in salivary, gastric, duodenal and pancreatic secretions were; 2.7 (0.4), 0.42 (0.12), 21 (5) and 8.5 (1.2) ng/ml, respectively. Chewing of Parafilm* significantly increased salivary but not gastric or duodenal EGF output while atropinisation led to the reduction in basal salivary and duodenal EGF output without affecting the increment in EGF release induced by chewing. Cigarette smoking caused a marked reduction in basal salivary and duodenal EGF output. Infusion of pentagastrin increased salivary and duodenal EGF output and this was blocked by the addition of somatostatin. Injection of secretin lead to an increase in pancreatic output of EGF. We conclude that in man the major sources of EGF are salivary glands, duodenum, and pancreas and that the release of EGF remains under neurohormonal control.     

Epidermal growth factor inhibits cysteamine-induced duodenal ulcers

The effect of the duodenal ulcerogen cysteamine on secretion of epidermal growth factor from Brunner's gland pouches was studied in the rat. Total output of immunoreactive epidermal growth factor was reduced to approximately 55%, compared with controls, 5 h after administration of cysteamine (300 mg/kg, s.c.). Furthermore, measurements on tissue extracts of the pouches revealed that 5 h after cysteamine treatment, Brunner's glands were depleted of epidermal growth factor. The effect on ulcer development of intraduodenally applied exogenous epidermal growth factor (1 micrograms/kg . h) also was studied. Luminal epidermal growth factor significantly inhibited the formation of cysteamine-induced duodenal ulcer, compared with controls receiving saline. The effect was not due to inhibition of gastric acid secretion or stimulation of duodenal bicarbonate secretion since the dose of epidermal growth factor used, when tested on chronic fistula rats, had no effect on acid secretion and did not influence bicarbonate secretion from Brunner's gland pouches. These results demonstrate that epidermal growth factor has a cytoprotective effect on the duodenal mucosa, and it is suggested that inhibition of synthesis and secretion of endogenous epidermal growth factor may be a pathogenetic factor in cysteamine-induced duodenal ulcer.     

Effects of somatostatin-14 on gastric and pancreatic responses to hormonal and neural stimulation using an isolated perfused rat stomach and pancreas preparation

To study the role of somatostatin in the regulation of pancreatic and gastric functions, a combined isolated rat stomach and pancreas preparation was developed. This model allowed simultaneous measurements of exocrine and endocrine secretion from the pancreas and gastrin secretion from the stomach. Somatostatin was applied either by a linear gradient or by constant infusion with one concentration in the presence of cerulein, secretin, electric vagal activity, or acetylcholine. Somatostatin did not influence exocrine pancreatic secretion irrespective of what substance was stimulated. In contrast, somatostatin significantly inhibited glucose-dependent insulin and gastrin secretion, either basal or stimulated by vagal activity or acetylcholine. Acetylcholine-induced gastrin secretion was more sensitive to inhibition by somatostatin than insulin. We conclude that in an isolated perfused organ system somatostatin has potent inhibitory effects on endocrine pancreas and stomach but has no effect on exocrine pancreatic volume and enzyme secretion.     

Release and binding of epidermal growth factor in the pancreas of rats.

Previous studies showed that EGF is produced by salivary and duodenal glands and released in saliva and duodenal secretion. Using specific radioimmunoassay of EGF, this study showed that the salivary glands and duodenal mucosa contain high levels of EGF, reaching, respectively, about 38 and 4 micrograms/g of tissue weight. EGF immunoreactivity was also found in high amounts in the pancreatic tissue (20 micrograms/g) and the pancreatic juice (32 ng/mL), where the content of EGF was found to increase in response to feeding, cholecystokinin, or bombesin and to decrease after the administration of atropine and somatostatin. Studies on the binding of EGF revealed that pancreatic acinar membranes possess the specific and saturable EGF receptors with a high affinity sites with Kd of about 4.3 nM and binding capacity of about 62 fmol/mg of protein, and with low affinity sites with Kd of 21 nM and binding capacity of about 180 fmol/mg of protein. The observed high content of immunoreactive EGF in the pancreatic tissue and the presence of high and low affinity binding sites for EGF in the pancreatic acinar membranes, as well as the high EGF output in the pancreatic juice and its alterations in response to hormonal and postprandial stimulation, suggest an important role of EGF in pancreatic physiology.     

Role of submandibular saliva and epidermal growth factor in gastric cytoprotection

The role of submandibular epidermal growth factor in protection of the gastric mucosa was investigated in rats. Removal of the submandibular glands and thereby submandibular epidermal growth factor (EGF) caused rats to develop gastric lesions (ulcerations and ulcers) after administration of the duodenal ulcerogen cysteamine. The median output of EGF in gastric juice was reduced from 45.6 pmol/12 h (total range 17.5-65.0) in unoperated controls to less than 0.06 pmol/12 h (total range less than 0.06-1.82) in rats given cysteamine after extirpation of the submandibular glands. The contents of EGF in the submandibular glands was unchanged during cysteamine treatment. Furthermore, the effects of intragastric instillation of exogenous EGF, infusion of saliva without EGF, and infusion of saliva with a high concentration of EGF on the development of cysteamine-induced gastric lesions were investigated in rats without submandibular glands. Exogenous EGF and saliva with a high but still physiological concentration of EGF significantly reduced the median area in the stomach displaying ulcers and ulcerations, whereas saliva without EGF had no effect. Although EGF is a known inhibitor of gastric acid secretion, the dose used in the present study had no effect on gastric acid secretion in chronic gastric fistula rats; removal of the submandibular glands also did not have any such effect. We conclude that exocrine secretion of submandibular EGF has a cytoprotective function in the stomach, an effect that may be physiological.     

Exploration of pancreatic exocrine function by continuous infusion of secretin

Summary A total of 47 subjects were given continuous intravenous infusion of secretin and an injection of pancreozymin. The results obtained from 20 control subjects showed that the secretin used in our study produced maximal alkaline secretion (711±166 µEq./min.) at a dose of 3 U./kg./hr. At this dose and up to 5 U./kg./hr., the infusion was perfectly tolerated by all the subjects.Compared with injection, the continuous-infusion method was not advantageous in measuring maximum bicarbonate concentration and lipase activity. By contrast, measurement of outputs showed great promise. Among the 4 parameters—volume, bicarbonate output, and maximum and mean lipase outputs—after pancreozymin, bicarbonate output seemed to be the most useful. Its measurement requires no more time than the standard secretin test, and this time could probably be reduced to 60 min. in most cases. Lipasic output is more variable, but undoubtedly will be of greater interest when it becomes possible to use higher doses of pancreozymin.The disadvantage of the continuous-infusion method lies in its great sensitivity—at present, it may be difficult to interpret the results in patients with nonpancreatic organic digestive diseases. Thus, more tests are needed to establish the statistical limits for the results in various types of disease.     

Effect of intraduodenal oligopeptide on rat pancreatic exocrine secretion and release of secretin and CCK]

We investigated whether intraduodenal (id) oligopeptide with three or four amino acids residues (pH 7.0) stimulates pancreatic exocrine secretion and release of endogenous plasma secretin and CCK in anesthetized rats. Id administration of oligopeptides in three doses (25, 100, 400 mg/hr) at a speed of 4 ml/hr resulted in dose-related increases in pancreatic secretion of pancreatic juice volume, bicarbonate, and amylase outputs (r = 0.598, 0.673, and 0.426, P less than 0.05 -- 0.001), and plasma concentrations of secretin and CCK (r = 0.743, 0.425, P less than 0.001 and 0.05). Intravenous administration of CCK-antagonist, CR1505 (5 mg/kg.hr) markedly inhibited oligopeptide-stimulated amylase output, but did not affect pancreatic juice volume and bicarbonate output. These results suggest that id oligopeptide increases pancreatic exocrine secretion and releases endogenous secretin and CCK.     

Effect of carbenoxolone on electrolyte output of salivary glands during nasogastric intubation

The effect of carbenoxolone on the output of salivary electrolytes stimulated by a nasogastric tube was studied in healthy volunteers. Mean ±se values for volume and electrolyte concentrations in control studies were volume 121±19 ml/hr, pH 7.39, Na⁺ 19 ±5, K⁺ 20 ±1, Cl^– 20±3, HCO ₃ ^– 18 mEq/liter and osmolality 71±8 mOsm/kg. Mean electrolyte outputs were quite large, Na⁺ 2.4, K⁺ 2.4, Cl^– 2.6, and bicarbonate 2.2 mEq/hr. Our control data indicate that in the course of gastric secretory testing the indwelling gastric tube stimulates salivary electrolyte output sufficient to interfere with accurate assessment of gastric ionic fluxes and gastric acid analysis. Therapeutic doses of carbenoxolone (100 mg three times daily) given orally for 3 weeks had no effect on salivary volume, electrolyte output, or concentration. We conclude that carbenoxolone has no minerallocorticoid effect on mixed salivary secretion. Attention to salivary secretion is recommended in all gastric secretory studies and is mandatory where low acid output is suspected.This work was supported in part by research grants from the U.S. Veterans Administration, and the National Health and Medical Research Council of Australia.     

Behavior of secretin-pancreozymin-stimulated pancreatic secretion, gallbladder contraction and choleresis after somatostatin administration

The action of somatostatin (250 microgram i. v. + 125 microgram/30 min i. v.) on pancreatic secretion was studied in 8 volunteers and on biliary secretion as measured through a T-tube in 7 volunteers during background stimulation of secretin and pancreozymin (1 U/kg/h). Additional injection of somatostatin was accompanied by a significant (p less than 0,05) decrease in volume and in bicarbonate output for a short time and a significant (p less than 0,05) decrease in trypsin, amylase and lipase output for the studied period. There was no decrease in volume output and bicarbonate concentration as measured in the T-tube. The effect of somatostatin in abolishing the pancreozymin induced contraction of the gallbladder was documented by sonography. The temporary reduction of volume and bicarbonate output in the duodenal lumen could be considered as an inhibitory effect of somatostatin on gallbladder contraction and not as an effect on the exocrine pancreas.