PubMed HealthSearch

Biomedical subjects

I De Dios

Publications and source records attributed to I De Dios.

At least 19 recordsLinked to original sources

Hydrocortisone induces an increase of amylase content in individual zymogen granules from rat pancreas.

This study was performed to evaluate the effects of different doses of hydrocortisone (1, 10 and 25 mg/kg/day) administered for 1, 3 and 8 days on pancreatic enzyme storage in rats. The enzyme content in both pancreas homogenates and in individual isolated zymogen granules (ZGs) was measured using standard biochemical assays and flow cytometry, respectively. Hydrocortisone did not alter the total amount of pancreatic DNA but increased the pancreas enzyme content in a time-dose-dependent way. Amylase activity was significantly increased after hydrocortisone administration at day +8 when 10 mg/kg/day was used, and from the first day of treatment when 25 mg/kg/day was administered. A significant increase in trypsin activity was also observed in response to 25 mg/kg/day of hydrocortisone but only from the third day of treatment onwards. As compared with control rats, chronic administration of either 1 or 10 mg/kg/day of hydrocortisone did not alter significantly either the size or the percentage of the two ZG subpopulations (Z1 and Z2) identified in the pancreas by flow cytometry; in addition, no significant changes were observed in the mean amylase content per individual granule, although its mean concentration increased in rats treated with 10 mg/kg/day for 3 and 8 days. Nevertheless, when 25 mg/kg/day of hydrocortisone were administered for 1 and 3 days, a significant increase in the proportion of Z1 ZGs was observed, which may be related to the formation of new and smaller ZGs. When a very high dose of hydrocortisone (25 mg/kg/day) was used, an overall increase in the pancreatic enzyme content related to an increase in the mean amylase content per individual ZG was observed; this effect was apparent from the first day of treatment in the Z1 subset of ZGs and from day +3 in the Z2 subpopulation. Only a high concentration of hydrocortisone was able to alter the enzyme storage process in individual zymogen granules, but they maintain a normal enzyme load at lower hydrocortisone doses.

Amylases

Adrenalectomy induces a decrease in the light scatter properties and amylase content of isolated zymogen granules from rat pancreas as analyzed by flow cytometry.

The effect of glucocorticoid deprivation induced in male rats by adrenalectomy on the pancreatic zymogen granules was studied. Zymogen granules were purified from control, sham-operated and adrenalectomized animals studied 1, 3 and 7 days after surgery. The zymogen granules were characterized by flow cytometry, and in each granule the size (based on the forward or low angle light scatter (FSC) parameter), membrane complexity (based on side or 90 degrees light scatter (SSC) parameter) and amylase content were evaluated. Amylase content/DNA ratio in pancreatic homogenates was also analyzed. The zymogen granules of the control rats were found to be distributed in two populations: a major one-R1 (95.45 +/- 1.21%)-containing zymogen granules with a smaller mean size and complexity, and a minor population-R2 (4.45 +/- 0.24%)-the granules of which had a mean size which was larger and more complex. At day +1 after adrenalectomy the zymogen granules were significantly (P < 0.05) smaller than those of control animals. The R2 zymogen granules were similar to those from R1 as regards their size, but were more complex, suggesting that the immediate effect of glucocorticoid deprivation is to induce a depletion of the larger granules presumably belonging to the R2 population. The amount of amylase per granule did not vary at day +1 after adrenalectomy, although the amylase content/size ratio per granule was significantly (P < 0.001) increased. This mechanism could be explained in terms of the existence of a bypass defined in the adrenalectomized animals between the granular content and cytosolic enzymes. Prolongation of the adrenalectomy period to 3 and 7 days resulted in a progressive increase in zymogen granule size and complexity, both parameters showing similar characteristics to those of the controls at day +7 after adrenalectomy. However, the percentage of zymogen granules within the R1 and R2 populations was clearly different from that of controls since the R2 population was much more numerous (11.25 +/- 0.75% and 15.25 +/- 1.15% (adrenalectomized rats at days +3 and +7 respectively) versus 4.45 +/- 0.24% (controls)). An increase in the content of amylase per DNA was observed in adrenalectomized rats at day +1 although this transient effect cannot be related to glucocorticoid deprivation because it was also observed in sham-operated rats (day +1). However, a significant reduction, nearly 64%, in the amylase content/DNA ratio is produced by the absence of glucocorticoids 7 days after adrenalectomy and this is associated with a reduction in the content of amylase in each individual zymogen granule which reaches a minimum 3 days after adrenalectomy. It should be noted that, despite this, the enzyme concentration in each granule remains constant as there is a parallel decrease in the zymogen granule amylase content and size.

Adrenalectomy

Effects of the cholecystokinin receptor antagonist L-364,718 on pancreatitis induced by a deficient in choline and supplemented with ethionine (CDE) diet in the rat.

The role of cholecystokinin (CCK) in the development of a necrotizing acute pancreatitis induced by a diet deficient in choline and supplemented with ethionine (CDE) has been evaluated in the rat by using a potent CCK receptor antagonist L-364,718. Acute pancreatitis was induced by administration of CDE diet for 14 days. L-364,718 administration was carried out by subcutaneous injections at dose of 0.1 mg/kg/day. Pancreatic exocrine secretion (flow, protein, amylase and trypsin outputs) in resting and under infusion of 1.25 microgram/kg/h of CCK-8 were used to evaluate the pancreatic functionality. Others parameters (serum amylase, percentage fluid in pancreas, haematocrit and mortality) evaluated the severity of pancreatitis. L-364,718 slightly reduced the mortality and the increases of percentage of fluid accumulated in pancreas in CDE diet acute pancreatitis. Basal and CCK stimulated pancreatic secretion was significantly depressed 36 hours after L-364,718 treatment. A slight response to CCK was observed. Nevertheless it was lower than usually observed in control rats. Our results demonstrate that in the rat, chronic L-364,718 treatment did not completely restore pancreatic activity in acute pancreatitis induced by CDE diet. Hence CCK cannot be considered as the main factor involved in the development of this pancreatitis model.

Acute Disease

Dose-response effects of VIP on the rabbit exocrine pancreatic secretion. Comparison with PACAP-27 actions.

A dose-response study of the effects of vasoactive intestinal peptide (VIP) on the exocrine pancreatic secretion of the rabbit has been made. Furthermore, the actions of VIP and pituitary adenylate cyclase activating peptide (PACAP) on the exocrine pancreatic secretion were compared at a similar molar dose. After the infusion of VIP a linear dose-response relationship for pancreatic flow rate and bicarbonate output, up to the dose of 4 micrograms/kg, was observed. VIP acts as a partial agonist of secretin, the rabbit pancreas being less sensitive to VIP compared with other mammals. Moreover, VIP did not significantly stimulate the pancreatic protein output. PACAP stimulated the hydroelectrolyte fraction of the exocrine pancreatic secretion in a similar manner to that of VIP. Unlike what was observed with VIP, PACAP, on the same molar basis, significantly stimulated the protein and amylase outputs. Furthermore, PACAP releases VIP, so that the action of PACAP on the hydroelectrolyte fraction may be partially mediated by VIP; on the other hand, VIP is not involved in the effect of PACAP on the pancreatic enzyme secretion of this species.

Amylases

Effect of L-364,718 (CCK receptor antagonist) on exocrine pancreatic secretion of hydrocortisone-treated rats.

Exocrine pancreatic function was studied in control rats and animals subjected to treatment with hydrocortisone (10 mg/kg/day) over 7 days. In both cases, the cholecystokinin (CCK) receptor antagonist L-364,718 (0.1 mg/kg/day) had been administered. The administration of this nonpeptide CCK antagonist significantly reduced the basal pancreatic flow of secretion and enzymes in the control rats but did not cause a decrease in pancreatic weight. From this it may be inferred that factors other than CCK are able to exert the trophic effects of this hormone. Also, the processes of enzyme synthesis and storage, which contribute to maintaining the weight of the organ, continued to occur under conditions of deprivation of the action of endogenous CCK because pancreatic secretion in response to the infusion of CCK was similar in the animals treated and not treated with L-364,718. In contrast, no significant changes were observed either regarding pancreatic weight or basal pancreatic secretion in the animals treated with hydrocortisone and simultaneously with L-364,718, as compared with the rats treated only with hydrocortisone. This points to a predominant effect of glucocorticoids on the action of CCK during the phases of enzyme synthesis and storage, accompanied by a blockade of exocytosis due to hydrocortisone treatment. The secretagogue effect of CCK alone becomes possible with high levels of this hormone of exogenous origin, whose interaction with its specific receptors is in turn positively modulated by an excess of glucocorticoids.

Amylases

Therapeutic and protective effect of subcutaneous injections of L-364,718 on caerulein-induced acute pancreatitis.

The prophylactic and therapeutic effects of a potent cholecystokinin (CCK) receptor antagonist, L-364,718, on acute pancreatitis induced by caerulein were evaluated, analyzing morphologic and functional pancreatic parameters jointly. Edematous pancreatitis was induced by four subcutaneous injections of caerulein (20 micrograms/kg) in rats at 1-h intervals. Prophylactic administration of L-364,718 (0.1 mg/kg) prevented rise in serum amylase levels, interstitial edema, vacuolization, and impairment of pancreatic enzyme secretion that accompany caerulein-induced acute pancreatitis. After 7 days, a spontaneous regression of the morphologic alterations caused by caerulein-induced acute pancreatitis occurs; however, recovery of the secretory function of the pancreas was only reached after this period of time when L-364,718 was administered therapeutically (0.1 mg/kg/day). Prophylactically or therapeutically administered, L-364,718 exerts a beneficial effect on caerulein-induced acute pancreatitis, indicating that CCK (exogenous or endogenous) plays an important role in the development of this pathology.

Acute Disease

Exocrine pancreatic response to intraduodenal fatty acids and fats in rabbits.

1. The exocrine pancreatic secretion in rabbits after intraduodenal perfusion of, respectively, a short-chain fatty acid (propionate), a long-chain fatty acid (oleate) and a neutral fat (intralipid) has been studied. 2. Only the neutral fat and the long-chain fatty acid stimulated the hydroelectrolyte fraction of exocrine pancreatic secretion in a dose-related fashion, but this response was not mediated by secretin release. 3. Acinar secretion was more effectively stimulated after propionate perfusion than by oleate and this was governed by different mechanisms. 4. Neutral fat, without predigestion, stimulated the pancreatic enzyme secretion in the rabbit.

Animals

Effect of L-364,718 (CCK receptor antagonist) on exocrine pancreatic secretion of adrenalectomized rats.

The exocrine pancreatic secretion of control and adrenalectomized rats treated with L-364,718 was studied. The blockade of the action of endogenous cholecystokinin (CCK) in the control animals was reflected in a reduction of basal pancreatic secretion. This effect was reversed by the administration of submaximal doses of CCK-8. Under treatment with L-364,718, no decrease in pancreatic weight was observed. From this it may be deduced that factors other than CCK act on the pancreas to maintain the processes of enzyme synthesis and storage. Adrenalectomy reduced the weight of the pancreata; the main cause of this was the depletion of zymogen granules, which has been reported in adrenalectomized rats and is accompanied by a reduction in enzyme secretion, especially that of amylase. By contrast, the administration of L-364,718 from the very first moments after adrenalectomy leads to a retention of pancreatic enzymes. This effect is reflected in the maintenance of pancreatic weight and a secretory capacity in response to submaximal doses of CCK-8 similar to that of nonadrenalectomized animals. Accordingly, in the absence of glucocorticoids, endogenous CCK dominates the secretory process because when the action of this secretagogue is prevented by the administration of L-364,718, the discharge of proteins able to be secreted in response to a suitable stimulus is inhibited.

Adrenalectomy

Acinar cholecystokinin (CCK) receptors in the exocrine pancreas of the rat: effect of adrenalectomy.

The binding of [125I]BH-CCK-8 to membranes of acinar cells from rats at 6 and 21 days after adrenalectomy was studied. The optimum conditions of time and temperature were previously established as being 120 min and 30 degrees C. Under these conditions, the membranes of the adrenalectomized animals of both groups (6 and 21 days) bound more radioligand than those from control rats. However, a qualitative study of the binding showed that the affinities of binding were much lower; in particular, the high affinity receptors had a Kd of 0.94 +/- 0.33 nM in the controls and this was 14.9 +/- 1.29 nM in the 6-day adrenalectomized animals, although the maximum binding capacity did not vary significantly. However, in the case of the low affinity receptors, there was a gradual increase in the maximum binding capacity as the time after adrenalectomy progressed: 717 +/- 121, 1,987 +/- 183, and 10,175 +/- 862 fmol/mg for the control, 6-day, and 21-day adrenalectomized rats, respectively. In the latter situation, the high affinity receptors completely disappeared. These results, which coincide with a marked deficit in protein secretion already described in adrenalectomized rats, can be accounted for in terms of the possible negative cooperativity exerted by the low affinity receptors on the high affinity hormone-receptor complex, the protein secretion of the acinar cells normally mediated by the high affinity receptors becoming paralyzed.

Adrenalectomy

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.

Acute Disease

Beta 1 and beta 2 adrenergic agonists in exocrine pancreatic secretion in the rabbit.

The effect of Dobutamine (a beta 1-adrenergic agonist) and Terbutaline (a beta 2-adrenergic agonist) on exocrine pancreatic secretion was studied in anaesthetized rabbits, simultaneously controlling pancreatic blood flow and blood pressure. The secretion of fluid and ions (bicarbonate, sodium and potassium) was unaffected by the infusion of Dobutamine (8 micrograms.kg-1.min-1) or Terbutaline (10 micrograms.kg-1.min-1). Neither were pancreatic blood flow or mean blood pressure altered. Dobutamine or Terbutaline depress the function of the acinar cells, amylase secretion being more affected by the action of Terbutaline. The results show that beta 1 and beta 2-adrenergic stimulation has no effect on the ductular cells but does decrease the secretion by the acinar cells.

Adrenergic beta-Agonists

Exocrine pancreatic response to isoproterenol in the rabbit under secretin and CCK stimulation.

The effect of isoproterenol (0.3 micrograms/Kg/min) on exocrine pancreatic secretion stimulated by secretin (1. C.U./Kg/h) and the octapeptide of cholecystokinin (OP-CCK 0.15 Ivy dog units/Kg/h) was studied in anaesthetized rabbits. Pancreatic blood flow remained unaffected by the action of isoproterenol; however, the administration of the beta-adrenergic agonist over an infusion of secretion an OP-CCK led to an increase in pancreatic juice flow (approximately 146% for secretin and 125% for OP-CCK) and HCO3-output (approximately 194% for secretin and 150% for OP-CCK), a potentiation being observed between the effects of secretin and isoproterenol. This kind of response, independent of vascular changes, is due to a direct action of isoproterenol on the ductular cells. Isoproterenol interferes with the action of secretin and OP-CCK in that it partially inhibits the stimulatory action of OP-CCK on the enzymatic secretion of the acinar cells in the rabbit pancreas and at the same time protein secretion decreases in the presence of secretin.

Acid-Base Equilibrium

Influence of glucocorticoids on the secretion of pancreatic juice in the rat.

The influence of adrenalectomy and hydrocortisone treatment on the exocrine pancreatic secretion has been studied in anaesthetized rats. In the adrenalectomized animals Na+ administered in the saline solution provided for drinking was able to maintain standard sodium levels in serum. In these animals an increase of Na+ secretion in pancreatic juice was observed. Furthermore, the osmotic effect created by the increase in Na+ would account for the increase in pancreatic flow. In these adrenalectomized rats, an increase in K+ output is observed, which can be explained by the high K+ concentrations in serum. Likewise adrenalectomy decreased pancreatic enzyme secretion and produced a loss in weight of the organ that is accounted for by a lack of glucocorticoids. Hydrocortisone administration did not affect neither the secretion nor the weight of the pancreas of the control rats but the hormone proved to be effective in adrenalectomized rats producing a pancreatic secretion close to normal, balancing the secretory rate of water, Na+ and K+, completely restoring total protein secretion and the weight of the pancreas but amylase secretion in part only. It is therefore concluded that the weight of the pancreas and its exocrine secretion are clearly influenced by adrenalectomy and by substitution therapy with hydrocortisone. The administration of this hormone (25 mg.kg-1.day-1 along 6 days) did not affect intact animals.

Adrenal Glands

Separation of human lymphocyte subclasses by rosettes with latex-lectin particles.

The specificity and affinity of eight lectins (concanavalin A, L. culinaris, P. sativum, phytohemagglutinin P, D. biflorus, soybean agglutinin, T. purpureus, and T. vulgaris) to B, T, T gamma, and T mu lymphocytes from the blood of normal subjects were determined. Lectins attached to latex particles were used to evaluate the binding of each lectin to individual cells. The rosette percentage found in each lymphocyte population expresses the specificity index and the specific sugar concentrations needed to decrease the rosette percentage by 50% is taken as the affinity index. B Lymphocytes showed a major subclass, with respect to T lymphocytes, with receptors for WGA, SBA, D. biflorus, L. culinaris, and P. sativum lectins. In contrast, T lymphocytes exhibit a greater number of cells with specific receptors for Con A, T. purpureus, and PHA lectins than B lymphocytes, the T gamma subpopulation being responsible for the specificity of the first two lectins and the T mu subpopulation for the PHA lectin.

B-Lymphocytes

Studies on platelet surface carbohydrates in normal and uraemic platelets using 125I-labelled lectins.

Binding studies with six different purified 125I-labelled lectins, concanavalin A (con A), wheat germ agglutinin (WGA), Ricinus communis agglutinin II (RCA II), Dolichos biflorus (DB), Tetranolobus purpureus (TP) and P-phyto-hemagglutinin (P-PHA), were used to investigate the surface topography of carbohydrates in platelets from uraemic and normal subjects. Compared with normal the uraemic platelets, bear significantly decreased (more than 2.5-fold) numbers of receptors for P-PHA (N-acetyl D-galactosamine specificity) and Con A (specificity glucose, mannose). The number of WGA, RCA, II, DB and TP receptors in uraemic platelets did not differ from the number in normal platelets. Binding studies with 125I-labelled lectins provide further evidence of molecular defects in uraemic platelets. Moreover, this method might provide a fast and reliable technique for identifying abnormalities in the surface topography of carbohydrates on platelets in several pathological states.

Adult