M.A. López

Basolateral anion transport mechanisms underlying fluid secretion by mouse, rat and guinea-pig pancreatic ducts.

Fluid secretion by interlobular pancreatic ducts was determined by using video microscopy to measure the rate of swelling of isolated duct segments that had sealed following overnight culture. The aim was to compare the HCO3− requirement for secretin‐evoked secretion in mouse, rat and guinea‐pig pancreas. In mouse and rat ducts, fluid secretion could be evoked by 10 nm secretin and 5 μm forskolin in the absence of extracellular HCO3−. In guinea‐pig ducts, however, fluid secretion was totally dependent on HCO3−. Forskolin‐stimulated fluid secretion by mouse and rat ducts in the absence of HCO3− was dependent on extracellular Cl− and was completely inhibited by bumetanide (30 μm). It was therefore probably mediated by a basolateral Na+–K+–2Cl− cotransporter. In the presence of HCO3−, forskolin‐stimulated fluid secretion was reduced ∼40% by bumetanide, ∼50% by inhibitors of basolateral HCO3− uptake (3 μm EIPA and 500 μm H2DIDS), and was totally abolished by simultaneous application of all three inhibitors. We conclude that the driving force for secretin‐evoked fluid secretion by mouse and rat ducts is provided by parallel basolateral mechanisms: Na+–H+ exchange and Na+–HCO3− cotransport mediating HCO3− uptake, and Na+–K+–2Cl− cotransport mediating Cl− uptake. The absence or inactivity of the Cl− uptake pathway in the guinea‐pig pancreatic ducts may help to account for the much higher concentrations of HCO3− secreted in this species.

Cerulein-induced acute pancreatitis in the rat : study of pancreatic secretion and plasma VIP and secretin levels

A study was made with different doses of cerulein (2, 4, 10 and 20 μg/kg) administered subcutaneously to rats by four injections at intervals of 1 hr; the aim of this work was to study exocrine pancreatic secretion of the rat under cerulein-induced acute pancreatitis, analyzing enzyme and hydroelectrolyte secretion of pancreatic juice. A further aim was to study the relationship between the dose of cerulein and the plasma levels of peptides controlling hydroelectrolyte secretion of the pancreas, like secretin and vasoactive intestinal peptide (VIP). At the lowest dose schedule, the amounts of total protein and enzymes (amylase and trypsin) in pancreatic juice decreased significantly, plasma amylase increased, and the pancreas became edematous. Higher doses magnified these effects. By contrast, ductular function (flow and HCO3−) was well preserved in ceruleintreated rats, and this was probably due to the significant increase in plasma levels of immunoreactive secretin whereas VIP levels were unchanged. The secretin released by treatment with cerulein is able to palliate the lack of flow from acinar origin that is affected in the process of acute pancreatitis, being a beneficial response to the cerulein treatment.

Exocrine pancreatic response to CCK in rats after long-term hydrocortisone administration

The effects of prolonged administration of hydrocortisone (10 mg/kg/day) on exocrine pancreatic secretion were examined, analyzing the quantitative and qualitative variations in secretion at 7, 18 and 30 days of treatment. The weight of the pancreata was found to increase during the period of hydrocortisone treatment. After the 7th day of treatment the hormone significantly decreases basal exocrine pancreatic secretion, maintaining similar values up to the 30th day of treatment. Hydrocortisone enhances the pancreatic response to CCK since the percent increase in acinar secretion under stimulation compared to basal secretion surpassed control levels in all cases. However, the inhibitory effect on secretion shown by hydrocortisone opposes and surpasses the stimulatory effect of the secretagogue, secretion being reduced in the treated animals. Hydrocortisone especially affected the amylase fraction of the juice, to a larger extent inhibiting the isoenzyme A1, with an IEP of 8.5 when the animals were treated over 30 days; thus, a considerable reduction was observed in the amylase secreted both in resting conditions and under stimulation with CCK. There is a possibility that chronic treatment with glucocorticoids may sensitize the acinar cells, alter the composition of the pancreatic juice and inhibit secretion, effects that may involve pancreatic dysfunction.

Effect of Alloxan Diabetes on Exocrine Pancreatic Secretion in the Anesthetized Rabbit

SummaryThe influence of the endocrine pancreas on the exocrine pancreatic secretion of both electrolytes and enzymes was studied in rabbits made diabetic by alloxan administration. No alterations were observed in the flow of pancreatic juice. Bicarbonate concentrations were considerably increased, whereas chloride concentrations were clearly reduced in the alloxan-diabetic rabbits compared with the control animals. Insulin treatment, restored anion levels to normal. Similar, although less pronounced changes were seen in the output values of bicarbonate and chloride. There were no significant differences between the control and alloxan-treated animals in the output of sodium and potassium; however, potassium concentrations exhibited a significant rise both in untreated and insulin-treated diabetic rabbits compared with the controls. Total protein and amylase secretion decreased markedly in the diabetic animals. The secretion of amylase was not brought back to control values by additional administration of insulin. These results suggest that the endocrine pancreas plays an extremely important role in exocrine pancreatic function.

Effect of secretin on pancreatic juice proteins in caerulein-induced acute pancreatitis in the rat

Nine hours after the start of treatment with caerulein in rats, an increase in the weight of the pancreas and an increase in serum amylase levels were observed. Likewise, a significant increase in endogenous secretin occurred in rats with acute pancreatitis. A dramatic reduction in the secretion of total protein and amylase was also observed. A partial recovery of this latter effect was achieved after an infusion of high doses of secretin. Under our experimental conditions, the volume of secretion did not vary in caerulein-treated rats wtih respect to controls, either in resting conditions or under secretin stimulation, which indicates that the ductular cells were not significantly affected. Isoelectrofocusing (IEF) and crossed-immunoelectrophoresis (CIE) studies revealed important alterations in the proteins of the pancreatic juice of rats with caerulein-induced acute pancreatitis. Trypsinogen appeared to be particularly affected, showing an increase in the T2 acidic form with an IEP of 4.4 and a decrease in the basic form T3 with an IEP of 8.0, which splits in other forms with a clear antigenic community. A hydrolase was also observed with an IEP of 6.2. In this sense, secretin administration may also be said to induce a significant improvement in established acute pancreatitis, since it tended to normalize the structure and proportion of the proteins secreted.

Duodenal Alkalinization Releases Secretin and Vasoactive Intestinal Polypeptide and Stimulates Exocrine Pancreatic Secretion in the Anesthetized Rat

The effect of various intraduodenal alkaline solutions (0.1 M NaHCO3, 0.1 M Na2CO3 and 0.025 M NaOH) on exocrine pancreatic secretion and the release of two peptides, secretin and vasoactive intestinal polypeptide, was studied in anesthetized rats. The flow rate of the pancreatic juice was stimulated up to a maximum of 179, 158 and 180% and the protein output up to 181, 131 and 162% (compared with basal) after duodenal perfusion of, respectively, 0.1 M NaHCO3, 0.1 M Na2CO3 and 0.025 M NaOH. Maximum increases in portal plasma secretin concentrations of 143, 146 and 190% and maximum increases in VIP of 116, 155 and 147% after, respectively, intraduodenal 0.1 M NaHCO3, 0.1 M Na2CO3, and 0.025 M NaOH were found. In conclusion duodenal alkalinization in the rat produces a pancreatic exocrine secretory response that may be partially ascribed to the effect of secretin and VIP.

Caerulein-induced acute pancreatitis in the rat. Pancreatic secretory response to cholecystokinin

The response of pancreatic exocrine secretion to cholecystokinin (CCK), has been studied in experimental acute pancreatitis induced in rats by supramaximal doses of caerulein. Several doses of caerulein were used (4, 20 and 40 micrograms/Kg) and each one was administered by four subcutaneous injections over 3 h at hourly intervals. Pancreatic juice was collected 9 h after the first injection. The caerulein-treated animals showed a statistically significant increase in serum amylase levels. Secretory activity of ductular cells remained unchanged in all the caerulein-treated animals, but total protein and amylase secretion decreased significantly at all the caerulein doses used, both in resting conditions and under stimulation with CCK (1.25 micrograms/Kg/h). Despite this the acinar cells of rats treated with the lowest dose of caerulein retained a certain degree of secretory function since amylase activity in pancreatic juice was greater than in other groups of rats treated with higher doses of caerulein. Moreover, the percentage of increase observed in total protein and amylase in response to CCK respect to basal secretion is similar to that of the untreated animals. At higher doses (20 and 40 micrograms/Kg) the secretory capacity in response to CCK was inhibited. Therefore CCK administration in slight acute pancreatitis could be used as a therapy since it favours the secretion of pancreatic enzymes at percentual levels similar to those of the controls.

Saline Infusion Through the Pancreatic Duct Leads to Changes in Calcium Homeostasis Similar to Those Observed in Acute Pancreatitis

This work focuses on studying the early events associated with pancreatic damage after retrograde infusion through the pancreatic duct in rats. We have analyzed changes in calcium homeostasis and secretory response in pancreatic acini from rats with taurocholate-induced acute pancreatitis. Moreover, in order to test whether pancreatic duct manipulation can trigger damage inside pancreatic acinar cells, we have studied both parameters in acini from animals infused with saline. Our study demonstrates that taurocholate causes evident damage to acinar cells, impairing both calcium homeostasis and secretory response to CCK. In saline, a significant decrease in calcium cytosolic response to CCK was observed. Calcium disturbances similar to those observed in acute pancreatitis appear before secretion blockade and inflammation processes in saline treated rats. These results could be interesting since pancreatitis is associated to clinical procedures that require duct manipulation such as endoscopic retrograde cholangiopancreatography.

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.

Effect of high fiber intake on pancreatic lysosomal stability in ethanol-fed rats 1

Abstract Chronic ethanol consumption increases the fragility of pancreatic lysosomes, but the effect of a high fiber intake, alone or combined with alcohol abuse, on the lysosomal stability has not been studied. Furthermore, it is not yet known whether these treatments could predispose the exocrine pancreas to a greater damage after cerulein-induced acute pancreatitis. Cytosolic specific activity of three lysosomal enzymes, N-acetyl-β- d -glucosaminidase (NAG), cathepsin B, and β-glucuronidase were measured, as an index of lysosomal stability in pancreas from control rats and rats under chronic alcohol and/or high fiber intake. Cathepsin B is the only enzyme with significantly increased specific activity after chronic ethanol consumption and, moreover, its specific activity undergoes the highest increase after cerulein-induced acute pancreatitis, in all the groups of rats, when compared with the remainder enzymes. When pancreatitis was induced by cerulein, the combination of chronic alcohol and high fiber intake produces a significant decrease in the cytosolic specific activity of N-acetyl-β- d -glucosaminidase and β-glucuronidase when compared with chronic alcohol alone. Our results suggest that fiber partially avoids the damage of ethanol on pancreatic lysosomes, reducing the effects of pancreatitis.