PubMed Health⌕ Search

Biomedical subjects

R D Duan

Publications and source records attributed to R D Duan.

48 records · Page 3Linked to original sources

Effect of insulin administration on contents, secretion, and synthesis of pancreatic lipase and colipase in rats.

After insulin administration in vivo, changes in pancreatic lipase, colipase and amylase contents and outputs were assayed and quantitatively compared. The incorporation of [35S]cysteine into individual enzymes was measured. The mRNA coding for lipase and amylase were determined by dot-blot hybridization. It was found that insulin dose-dependently decreased lipase and colipase contents, but only slightly decreased amylase content. Four hours after insulin administration (0.5 U/100 g), the contents of lipase and colipase decreased 80 and 72%, respectively, while amylase content decreased only about 25%. The decrease in amylase content was accompanied by a 21% increase in its output. The outputs of lipase and colipase only increased transiently and then sharply decreased to a level much lower than control. Total outputs of lipase and colipase could not quantitatively explain the great loss of lipase and colipase contents caused by insulin administration. After insulin injection, the incorporation of [35S]cysteine into amylase increased by 21%, whereas incorporation into lipase and colipase decreased by 18 and 25%, respectively. Dot-blot hybridization with cDNA probes revealed that lipase mRNA decreased by 50% 4 h after insulin administration, whereas mRNA for amylase did not significantly change. The results indicate an inhibitory effect of insulin administration on synthesis of pancreatic lipase and colipase, with the inhibition of lipase synthesis being at pretranslational level.

Amino Acids↗

The anticoordinate changes of pancreatic lipase and colipase activity to amylase activity induced by adrenalectomy in normal and diabetic rats.

The effect of adrenalectomy (Adx) on pancreatic lipase and colipase activities in normal, streptozotocin-induced diabetic, and obese Zucker rats was studied. It was found that Adx induced a rapid decrease in amylase activity, a rapid increase in lipase activity, and a slow increase in colipase activity in normal rats. Twenty days after Adx, the levels of amylase activity were 70% lower than control, whereas the activities of lipase and colipase were 55 and 25% higher than control, respectively. The replacement of hydrocortisone normalized the changes in the three enzyme activities. Injection of insulin partially corrected the amylase activity, but had no significant effect on the activities of lipase and colipase. In streptozotocin-induced diabetic rats and obese Zucker rats, amylase activity was decreased, and lipase and colipase activities increased, compared to normal rats. These changes of enzyme activity were further enhanced by Adx. It is concluded that Adx results in an anticoordinate change in pancreatic lipase, colipase, and amylase activities, suggesting that glucocorticoid also influences the synthesis of the pancreatic lipolytic enzymes.

Adrenal Glands↗

Altered synthesis of some secretory proteins in pancreatic lobules isolated from streptozotocin-induced diabetic rats.

The in vitro incorporation of [35S]cysteine into lipase, colipase, amylase, procarboxypeptidase A and B, and the serine proteases and total proteins was studied in pancreatic lobules isolated from normal and diabetic rats with or without insulin treatment. The incorporation of [35S]cysteine into total proteins was 65% greater in pancreatic lobules from diabetic animals than from normal rats. The increased incorporation was partly reversed by insulin treatment (2 U/100 g/day for 5 days) of diabetic rats. The relative rates of biosynthesis for amylase and the procarboxypeptidases in diabetic pancreatic lobules were decreased by 75 and 25%, respectively, after 1 h of incubation, while those for lipase, colipase, and the serine proteases were increased by 90, 85, and 35%, respectively. The absolute rates of synthesis for these enzymes changed in the same direction as the relative rates in diabetic lobules, except that for the procarboxypeptidases, which did not change. The changed rates of biosynthesis for the pancreatic enzymes were reversed by insulin treatment of the diabetic rats. Kinetic studies showed that the incorporation of [35S]cysteine into amylase, lipase, and colipase was linear until up to 2 h of incubation in normal pancreatic lobules, while in the diabetic lobules the incorporation into lipase and colipase was accelerated, reaching a plateau level already after 1 h of incubation. It is concluded that the biosynthesis of pancreatic secretory proteins in diabetic rats is greatly changed both in terms of quantity and kinetics.

Animals↗

Increase in pancreatic lipase and trypsin activity and their mRNA levels in streptozotocin-induced diabetic rats.

The pancreatic lipase and trypsin activities in streptozotocin-induced diabetic rats were determined as well as the relative levels of mRNA coding for these proteins. It was found that after development of diabetes, the activities of pancreatic lipase and trypsin were significantly increased by 105% and 52%, respectively, accompanied by an increase in the levels of lipase and trypsinogen mRNA by 98% and 49%, respectively, while amylase activity and its mRNA were significantly decreased. The alteration of lipase, trypsin, and amylase activities and their mRNA in diabetic rats were all normalized by replacement of insulin. It is concluded that in the diabetic situation, the pretranslational control for pancreatic lipase and trypsinogen is stimulated, resulting in high levels of these enzymes in the diabetic rat.

Amylases↗

Pancreatic lipase and colipase activity increase in pancreatic acinar tissue of diabetic rats.

We studied the lipase and colipase activity in pancreatic acinar tissue of insulin-deficiency and insulin-resistance obese Zucker rats (fa/fa). After injection of streptozotocin (STX 75 mg/kg) in normal Sprague-Dawley rats, the activity of lipase and colipase in pancreatic acinar tissue was increased by approximately 100%, the increase in colipase occurring 3 days later than that of lipase. At the same time, the amylase activity was decreased by 98%. Injection of alloxan (125 mg/kg) induced a similar change of pancreatic enzyme pattern, with amylase activity strongly reduced by 79% and activity of lipase and colipase increased 20.5 and 18.6%, respectively. Correction of the diabetic state with insulin (1 U/100 g/day) reversed the activity of these enzymes to their prediabetic levels. Administration of insulin (6 U/100 g/day) to normal Sprague-Dawley rats increased the activity of amylase as well as lipase and colipase, whereas injection of glucagon (0.3 mg/100 g/day) decreased the activity of amylase and colipase but had no significant effect on lipase activity. In the obese Zucker rats (fa/fa), the activity of lipase and colipase at onset of obesity (5 weeks of age) was lower than that in their lean littermates (fa/o). Thereafter the activity of the two proteins increased with age, being 40% higher in the fa/fa rat than in the fa/o rat at age 7 weeks. During the same period, amylase activity decreased. These results indicate that pancreatic lipase and colipase activity are increased following either insulin deficiency or insulin resistance in rats by a mechanism related to the changed levels of insulin.

Amylases↗

Effects of verapamil on amylase release from rat pancreatic acini and its relation to calcium fluxes.

The effects of verapamil on amylase release and Ca2+ fluxes from rat pancreatic acini have been studied. Verapamil at concentrations above 10 microM dose-dependently inhibited amylase release stimulated by carbachol, but enhanced the amylase release stimulated by cholecystokinin (CCK) and secretin. Verapamil had no significant effect on calcium uptake induced by carbachol or CCK, but significantly inhibited Ca+2 efflux caused by carbachol and slightly increased that caused by CCK. In a Ca2+-free, EDTA-containing medium, the increase in cytoplasmic free Ca2+ caused by carbachol was significantly inhibited by verapamil. Verapamil alone up to 400 microM had no effect on the release of lactic dehydrogenase. In conclusion, the effect of verapamil on amylase release from rat pancreatic acini differs depending on the type of secretagogue used to stimulate amylase release. This effect is not related to blockage of Ca2+ uptake, indicating another mechanism of verapamil on pancreatic acini.

Amylases↗

Calcium fluxes caused by bile salts in rat pancreatic acini.

The effect of bile salt on Ca2+ uptake, Ca2+ efflux, and cytosolic free Ca2+ concentration ([Ca2+]i) in rat pancreatic acini has been studied. The dihydroxy bile salts, taurodeoxycholate (TDC) and taurochenodeoxycholate (TCDC), were found to accelerate Ca2+ uptake. Dihydroxy bile salt and the trihydroxy bile salt taurocholate (TC) stimulated Ca2+ efflux from the acini. Verapamil increased the Ca2+ efflux induced by dihydroxy bile salts but did not influence the Ca2+ uptake. Under calcium-equilibrated conditions TDC and TCDC caused a quick net Ca2+ efflux, followed by an increase in Ca2+ uptake, whereas TC only caused a net Ca2+ efflux. TDC and TCDC, but not TC, increased the [Ca2+]i. This effect of TDC and TCDC was abolished in Ca2+-free EDTA-containing (0.2 mM) medium. The amylase release caused by bile salts was related in time with the Ca2+ fluxes. In conclusion, bile salts may change the Ca2+ homeostasis of pancreatic acini in different ways, depending on the type of bile salt. This change of Ca2+ homeostasis may play an important role in the bile salt-stimulated amylase release.

Amylases↗

Effects of extracellular calcium and magnesium on bile-salt-stimulated amylase release from rat pancreatic acini.

The effects of extracellular calcium and magnesium on bile-salt-induced amylase release from rat pancreatic acini have been studied. The amylase releases caused by from 0.25 mM to 1.0 mM taurodeoxycholate (TDC) and by taurochenodeoxycholate (TCDC) at a concentration of more than 0.75 mM were reduced by ethylenediaminetetraacetic acid (EDTA) and increased by verapamil. EDTA and verapamil had no significant effect on the taurocholate (TC) (1.0-5.0 mM)-stimulated amylase release. The inhibiting effect of EDTA began to appear already during the initial 5 min and was not parallel to any change of lactate dehydrogenase release. The TDC- and TCDC-stimulated amylase release was strongly dependent on the concentrations of extracellular calcium and was only weakly dependent on extracellular magnesium. The TC-induced amylase release was slightly increased only at high concentrations of calcium and magnesium. It is suggested that the mechanism of dihydroxy bile-salt-induced amylase release from rat pancreatic acini is different from that of trihydroxy bile salt. The dihydroxy bile-salt-stimulated amylase release is dependent on extracellular calcium but does not seem to be related to the uptake of calcium by the acini.

Amylases↗

Stimulatory effects of human pancreatic polypeptide on rat pancreatic acini.

The effect of human pancreatic polypeptide (HPP) on rat pancreatic acini has been studied. It was found that HPP stimulated amylase and lipase release from the acini. The secretory response of acini to HPP was dose-dependent in a sigmoidal fashion. Between 10(-9) M and 10(-8) M concentration of HPP there was a slow increase of enzyme release to about 40-60% over basal release. At concentrations of HPP above 10(-8) M there was a rapid increase of enzyme release, amounting to 4-6 times over basal release at 10(-6) M concentration of HPP. The potency of HPP compared to other secretagogues at 10(-7) M concentration was 45% of CCK, 60% of carbachol and 75% of secretin. HPP did not inhibit the effect of CCK, secretin and carbachol on amylase release. The amylase release stimulated by HPP was accompanied by an increase in 45Ca2+ efflux. Atropine or dibutyryl cyclic GMP did not influence the effect of HPP. It is concluded that HPP stimulates the release of enzymes from rat pancreatic acini and that Ca2+ may be a mediator for this secretion.

Animals↗

Effect of bile salt on amylase release from rat pancreatic acini.

The effect of different bile salts on amylase release from isolated rat pancreatic acini has been studied. The bile salt-stimulated discharge of amylase could be divided into three situations, depending on the concentration of bile salt. At low concentrations, between 1 and 100 X 10(-6)M, there was a slight increase in amylase secretion, 5-7% of total, which varied with the type of bile salt but was independent of the concentration of bile salt. The release of amylase stimulated by cholecystokinin, secretin, and carbachol was not affected by bile salts at this low concentration. At slightly higher concentrations, between 250 and 1000 X 10(-6)M, there was a large release of amylase, 10-40% of total, which was dependent on both type and concentration of bile salt. This release occurred specifically for amylase and was not followed by release of either membrane-bound dipeptidylpeptidase IV or intracellularly located lactic dehydrogenase. At higher concentrations, 2000-5000 X 10(-6)M, both amylase and dipeptidylpeptidase IV and lactic dehydrogenase were released, accompanying viability changes of the cells with uptake of trypan blue.

Amylases↗

Activation of neutral sphingomyelinase participates in ethanol-induced apoptosis in Hep G2 cells.

The mechanism underlying ethanol-induced apoptosis in liver cells is not clear. Sphingomyelin (SM) metabolism is a novel signal transduction pathway that has an impact on apoptosis in many cell types. We investigated whether the SM pathway is involved in ethanol-induced apoptosis in the liver. Hep G2 cells were treated with ethanol followed by assaying apoptosis, sphingomyelinase (SMase) activity, caspase-3 activity, and the changes of SM content in the cells. We found that ethanol dose-dependently increased apoptosis and the effect was accompanied by increases of caspase-3 activity and neutral SMase activity. At concentrations of 80 and 160 mM, ethanol significantly increased caspase-3 activity by 120% and neutral SMase activity by 24%. The activity of acid SMase was only slightly increased without statistical significance. C(2)-ceramide, the exogenous SM metabolite, mimicked the effects of ethanol on apoptosis and caspase-3 activation. When the SM content was determined 24 h after treatment with ethanol, its level was 15% lower than that of controls. The results indicate that metabolism of SM triggered by neutral SMase participates in ethanol-induced apoptosis in Hep G2 cells and activation of caspase-3 is involved in the apoptotic pathway.

Apoptosis↗