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Biomedical subjects

K Kitani

Publications and source records attributed to K Kitani.

At least 37 records · Page 2Linked to original sources
PubMed1991

Leupeptin causes an accumulation of lipofuscin-like substances in liver cells of young rats.

Leupeptin, a thiol protease inhibitor, has previously been shown to cause a dense accumulation of substances resembling age pigment and called ceroid-lipofuscin, in brain cells of young rats. Thus far, however, attempts to produce age pigments in hepatocytes of normal young rats with protease inhibitor(s) have not been successful. The present study provides the first demonstration that leupeptin induces lipofuscin-like substances in normal young rat hepatocytes. Male Fischer-344 rats (age 4-6 weeks) were continuously infused with leupeptin or saline i.p. for 2 weeks by an osmotic minipump (dosage, 1-50 mg/100 g per day). Liver tissues were then examined by light, fluorescence and electron microscopy. Both hepatocytes and non-parenchymal cells of livers treated with leupeptin, but not saline, showed a dense accumulation of pigments which stained deeply with toluidine blue, were PAS-positive and were brightly autofluorescent. After UV excitation the pigments had an emission spectrum with a broad peak at 480-540 nm extending to 650 nm resembling the spectrum of age pigment from livers of normal aged rats. Electron microscopic examination revealed numerous lipofuscin-like deposits with heterogeneous morphology in the cytoplasm of both hepatocytes and non-parenchymal cells; lipid and myelin-like bodies were also present in hepatocytes. The results indicate that the perturbation of proteolytic activity in liver by leupeptin causes an accumulation of substances which by several criteria resemble lipofuscin. These results thus provide further support for the 'Protease Inhibitor Model of Lipofuscin Formation' as well as a potential experimental model for studying hepatocellular aging processes.

Aging↗
PubMed1991

Absence of luminal bile increases duodenal content of cholecystokinin in rats.

The effects of the removal of bile from the proximal intestine on pancreas, plasma cholecystokinin (CCK) concentration, and duodenal content of CCK were examined in rats. Bile was excluded from the duodenum and introduced into the distal ileum through a silastic cannula for 7 days. Pancreatic juice was maintained to be normally secreted into the duodenum. After 7-day bile diversion, plasma CCK concentration and duodenal CCK content were significantly increased in bile-diverted rats. Trypsin content in the proximal intestine in bile-diverted rats was one-half that in control. Pancreatic wet weight, protein content, and DNA content in the pancreas were slightly increased, and lipase content was slightly decreased, by bile diversion, but none of these changes was statistically significant. Amylase content significantly decreased and chymotrypsin content significantly increased in bile-diverted rats. Intragastric administration of camostate (trypsin inhibitor) significantly increased plasma CCK concentration in both bile-diverted and control rats, and the net increase was much greater in bile-diverted rats than in control rats. In conclusion, bile diversion increased duodenal CCK content and increased the CCK response to luminal stimulant.

Amylases↗
PubMed1991

Differences in tolerance to hypoxia/anoxia in mice of different ages.

Tolerance to hypoxic stress was examined in male C57BL mice of different ages through use of a hypoxic chamber (4% oxygen, 96% nitrogen). The basal survival times in a hypoxic chamber did not significantly differ between mice of different ages (6 wks, 8 wks, 10 months, 28 months). The pretreatment of antihypoxic prostaglandin I2 (PGI2) significantly prolonged the survival time in both adult (10-month-old and aged (30-month-old) mice, but the effect tended to be smaller in aged mice. The effect of sublethal hypoxia treatment on survival time in a hypoxic chamber was minimal in both adult and aged mice, but the effect tended to be smaller in aged mice. IP injection of sublethal doses of potassium cyanide (3.0 mg/kg) significantly prolonged the survival time in a hypoxic chamber when examined 10 min after the pretreatment in immature mice (6 wks), but in adult and aged mice, the survival time tended to be slightly shorter at that time than respective control values without pretreatment. The maximal survival time after KCN pretreatment was longest in immature mice and shortest in aged mice. The results suggest that the basal tolerance to hypoxic stress in mice is little affected by aging. However, the potential antihypoxic mechanisms appear to be progressively affected by aging.

Aging↗
PubMed1990

NB-598: a potent competitive inhibitor of squalene epoxidase.

NB-598, (E)N-ethyl-N-(6,6-dimethyl-2-hepten-4-ynyl)-3-[(3,3'-bith iophen-5-yl)methoxy]benzene-methanamine, was found to inhibit human microsomal squalene epoxidase (from Hep G2 cells) in a competitive manner. NB-598 inhibited cholesterol synthesis from [14C]acetate dose dependently in Hep G2 cells and increased the intracellular radioactivity of squalene. A single oral administration of NB-598 inhibited cholesterol synthesis from [14C]acetate in rats. Moreover, multiple oral administration of NB-598 to dogs decreased serum total and low density lipoprotein cholesterol levels and increased serum squalene levels. After termination of treatment, the reduced serum cholesterol and increased squalene levels returned to their control values.

Animals↗
PubMed1990

The importance of the C-terminal amide structure of rat pancreastatin to inhibit pancreatic exocrine secretion.

A C-terminal fragment of rat pancreatatin, a 26 residue peptide amide and a fragment without a C-terminal amide were synthesized by Fmoc-based solid phase methods and their biological activities were compared. The rat C-terminal fragment inhibited pancreatic exocrine secretions produced by the intravenous injection of 2-deoxy-D-glucose (a central vagal nerve stimulation), whereas the fragment without a C-terminal amide showed no effect on pancreas. These results indicate that the C-terminal amide of this peptide is necessary to reveal its biological activity.

Animals↗
PubMed1990

Rat pancreastatin inhibits both pancreatic exocrine and endocrine secretions in rats.

Effects of synthetic rat pancreastatin C-terminal fragment on both exocrine and endocrine pancreatic functions were examined in rats, in vivo and in vitro. Pancreastatin (20, 100 pmol, 1 nmol/kg/h) significantly inhibited CCK-8-stimulated pancreatic juice flow and protein output in a dose-related manner, in vivo. The inhibitory effect on bicarbonate output was not statistically significant. Pancreastatin did not significantly inhibit basal pancreatic secretions in vivo, and did not inhibit amylase release from the dispersed acini, in vitro. Insulin release stimulated by intragastric administration of glucose (5 g/kg) was significantly inhibited by pancreastatin (1 nmol/kg/h), in vivo. Plasma glucose concentrations were increased by pancreastatin infusion, but the increase was not statistically significant. Furthermore, pancreastatin inhibited insulin release from isolated islets, in vitro. Synthetic rat C-terminal pancreastatin fragment has bioactivities on both exocrine and endocrine pancreatic functions in rats.

Amylases↗
PubMed1990

Age-dependent decrease in the hepatic uptake of taurocholic acid resembles that for ouabain. A possible role of surface membrane protein mobility.

Isolated hepatocytes were prepared from Wistar-derived male rats of different ages (4, 12 and 27-29 months) by the collagenase perfusion method. The hepatic uptake rate of taurocholate (TC) for the saturable fraction was calculated by subtracting the non-saturable fraction from the total hepatic uptake. The Vmax and the apparent affinity constant Km were computed for the saturable fraction by means of non-linear regression. The Vmax (nmol/mg protein/min, mean +/- SE) for young rats (N = 6) was 2.15 +/- 0.11, whereas in old rats (N = 4) the value was 50% lower (1.16 +/- 0.11, P less than 0.005). In contrast, Km (microM) values were not significantly different between young (25.88 +/- 1.90) and old (30.34 +/- 4.96) rats. There was a significant inverse linear relationship (r = 0.79; P less than 0.01) between the age of rats and the uptake velocity (nmol/mg/mg protein/min) at 1 microM of TC, suggesting a steady and almost linear decrease of TC uptake velocity with age. The rate of decrease per month (2.1%) was quite close to the value for ouabain uptake (2.8%) previously found by the authors. Furthermore, a marked linearity was observed between the average values for TC uptake rates for three age groups and corresponding lateral diffusion constants of hepatocyte plasma membrane proteins previously obtained by the authors using the fluorescence recovery after photobleaching method. The results support our previous proposal that protein mobility within the hepatocyte surface membrane may play at least a partial role in regulation of carrier-mediated hepatocyte uptake functions for various materials.

Age Factors↗
PubMed1990

Comparative effects of simvastatin (MK-733) and pravastatin (CS-514) on hypercholesterolemia induced by cholesterol feeding in rabbits.

The preventive effects of simvastatin (MK-733) and pravastatin (CS-514), 3-hydroxy-3-methylglutarylcoenzyme A (HMG-CoA) reductase inhibitors, on hypercholesterolemia induced by 0.25% cholesterol feeding were compared in rabbits. MK-733 (6, 2 and 0.7 mg/kg) was found to prevent the increase in serum total cholesterol levels dose-dependently. High dose CS-514 (18 mg/kg) also limited the increase in the cholesterol levels, but medium (6 mg/kg) and low doses (2 mg/kg) of CS-514 were ineffective in preventing it. MK-733 inhibited the increase in VLDL and LDL cholesterol levels dose-dependently. MK-733 suppressed the increase in serum phospholipid levels. MK-733 inhibited the accumulation of cholesterol in the liver. The high dose of CS-514 also limited it. High dose MK-733 (6 mg/kg) reduced the cholesterol concentration in gallbladder bile. Neither MK-733 nor CS-514 affected bile acid excretion in the gallbladder bile. High dose MK-733 decreased the lithogenic index. MK-733 increased the number of LDL receptors, and high dose CS-514 also increased it. The suppressive effect of CS-514 on serum cholesterol levels at 18 mg/kg was found to be less than that of MK-733 at 0.7 mg/kg.

Animals↗
PubMed1990

Hepatic injury induced by bile salts: correlation between biochemical and morphological events.

Continuous intravenous infusion of taurochenodeoxycholate at a rate of 0.4 mumol.min-1.100 gm-1 for only 30 min in rats caused threefold to tenfold greater release of proteins (alkaline phosphatase, lactate dehydrogenase and albumin) into bile in comparison with animals infused with tauroursodeoxycholate at much higher rates (1.8 mumol.min-1.100 gm-1) for 2 hr. The simultaneous infusion of tauroursodeoxycholate and taurochenodeoxycholate (0.6 and 0.4 mumol.min-1.100 gm-1, respectively) for 2 hr prevented the marked biochemical changes in the bile induced by taurochenodeoxycholate for 15 to 60 min exhibited significantly more necrotic hepatocytes, especially in zone 1, in comparison with animals infused with tauroursodeoxycholate or a combination of taurochenodeoxycholate and tauroursodeoxycholate. A good correlation was observed between biochemical and morphological indices of bile acid-induced hepatocyte injury. These data suggest that (a) primary events induced by the acute infusion of toxic bile salts responsible for cholestasis include zone 1 hepatocellular necrosis and (b) this can be prevented by the simultaneous infusion of tauroursodeoxycholate.

Animals↗
PubMed1990

Hydrolysis and acyl migration of a catechol monoester of L-dopa: L-3-(3-hydroxy-4-pivaloyloxyphenyl)alanine.

Hydrolysis and acyl migration studies on L-3-(3-hydroxy-4-pivaloyloxyphenyl)alanine (1, NB-355), which produced long-lasting plasma L-dopa levels after oral dosing, have been conducted. Compound 1 exists as pure 4-O-pivaloyl-L-dopa in the solid state, but it converts rapidly to a mixture of the 3- and 4-O-isomers in solution. The rate of acyl migration increased with increases in pH and temperature, and the content of the 4-O-isomer in the equilibrium state was 53-59%. The hydrolysis rate of 1 to L-dopa (6) also increased with increases in pH and temperature, and accelerated steeply at neutral and alkaline pH. The rapid hydrolysis at neutral pH was not observed with O-pivaloyl-L-tyrosine (3), di-O-pivaloyl-L-dopa (4), or L-dopa methyl ester (5). Because of this chemical lability, 1 was hydrolyzed in rat plasma far faster than the other tested catechol esters. However, in rat intestinal homogenate at pH 6.0, 1 was hydrolyzed at the slowest rate among the tested esters, predominantly by a diisofluorophosphate (DFP)-sensitive esterase. Thus, 1 showed a unique in vitro profile on hydrolysis and acyl migration due to the existence of a neighboring hydroxyl group. The stability of 1 in the intestine might be essential for the long-lasting plasma L-dopa profile after oral dosing of 1.

Acylation↗
PubMed1990

Luminal bile regulates cholecystokinin release in conscious rats.

The effects of intraluminal bile on cholecystokinin release and pancreatic exocrine secretion were studied in conscious rats. Since it has been suggested that bile acid may influence pancreatic secretion indirectly by interacting with luminal protease activities, intraduodenal protease activities were eliminated by pancreatic juice diversion accompanied with simultaneous intraduodenal infusion of aprotinin. This treatment resulted in gradual increases in pancreatic juice flow, bicarbonate and protein outputs, and an increase in plasma cholecystokinin levels, reaching plateau levels 2 hr after the start of the treatment. When endogenous bile was excluded from the intestine, the pancreatic secretion and plasma cholecystokinin concentrations further increased. The intraduodenal infusion of sodium taurocholate during bile pancreatic juice diversion inhibited cholecystokinin release, while pancreatic protein output was only transiently decreased. The results indicate that bile in the duodenum directly regulates cholecystokinin release, probably through its major components, bile salts.

Animals↗
PubMed1990

Inhibitory effect of pancreastatin on pancreatic exocrine secretions. Pancreastatin inhibits central vagal nerve stimulation.

The effects of the C-terminal fragment of rat pancreastatin on exocrine pancreatic secretions induced by several neural stimulations [IV injection of 75 or 15 mg/kg of 2-deoxy-D-glucose (central vagal nerve stimulation), injection of 2 mg of cisapride (proposed to elicit acetylcholine release from cholinergic nerve ending), and infusion of 1 or 3 mg.kg-1.h-1 of bethanechol (direct stimulation of acinar cells)] were examined in conscious rats. Rats with external bile and pancreatic fistulae were used. All the stimulations caused significant increases in pancreatic exocrine secretions. Pancreastatin at 100 pmol.kg-1.h-1 inhibited pancreatic secretions stimulated by IV injection of 2-deoxy-D-glucose but not those induced by the infusion of bethanechol or the injection of cisapride. Because these findings showed that pancreastatin inhibited pancreatic secretions induced by central vagal nerve stimulation, the effect of pancreastatin on cholecystokinin-stimulated pancreatic secretions in vagotomized rats was examined. Pancreastatin at 100 pmol.kg-1.h-1 did not inhibit pancreatic secretions stimulated by cholecystokinin octapeptide at 100 pmol.kg-1.h-1 in conscious rats after bilateral truncal vagotomy. These results suggest that pancreastatin inhibits pancreatic exocrine secretions by inhibiting vagal efferent nerve activity.

Animals↗
PubMed1990

The order of hepatic cytotoxicity of bile salts in vitro does not agree with that examined in vivo in rats.

Using an enzyme release from isolated rat hepatocytes incubated with a bile salt as a marker, the cytotoxic order of bile salts was found to be taurochenodeoxycholate, glycochenodeoxycholate greater than tauroursodeoxycholate, glycoursodeoxycholate, cholate greater than taurocholate, glycocholate. Thus, the cytotoxicity of conjugates of ursodeoxycholate was greater than that of conjugates of cholate. However, these data do not agree with the order of cytotoxicity of these bile salts previously studied in vivo by the authors which demonstrated the least cytotoxic nature of conjugates of ursodeoxycholate.

5'-Nucleotidase↗
PubMed1990

Tauro beta-muricholate is as effective as tauroursodeoxycholate in preventing taurochenodeoxycholate-induced liver damage in the rat.

Cholestasis and enhanced biliary leakage of proteins such as lactate dehydrogenase (LDH) and albumin are known to be induced by infusions of relatively toxic bile salts such as taurocholate (TC) and taurochenodeoxycholate (TCDC). Tauroursodeoxycholate (TUDC) was previously shown to prevent these bile abnormalities when simultaneously infused (1-5). In the present study, we examined whether tauro beta-muricholate (T beta-MC) has a similar effect. The enhanced biliary excretion of LDH and albumin induced by the infusion of TCDC at a rate of 0.4 mumol/min/100 g was markedly prevented by the simultaneous infusion of T beta-MC or TUDC at a rate one-fourth that of TCDC. Increased LDH level in plasma and hemolysis caused by the infusion of TCDC were also reduced by either T beta-MC or TUDC. These results indicate that T beta-MC has a preventive effect on TCDC-induced hepatobiliary changes, which is as efficient as that of TUDC as shown previously, suggesting that the 7 beta-hydroxy group is important for this hepatoprotective effect. Furthermore, our results suggest that beta-muricholic acid may also have clinical value since current reports demonstrate a beneficial effect of ursodeoxycholic acid on a variety of cholestatic conditions, including primary biliary cirrhosis.

Animals↗
PubMed1990

Effect of protein-free diet on activities and subunits of glutathione S-transferase in livers of young and aged female rats.

Female Fischer-344 rats of different ages (8 and 25 months old) were fed a protein-free diet (PFD) for 7 days and refed a normal diet (ND) (23% protein) thereafter. Rats were killed immediately after the PFD was stopped (day 0) and at different time intervals during refeeding of a ND. Four subunits (1,2,3 and 4) and activities of glutathione S-transferases (GSTs) toward five different substrates, [styrene oxide (STOX), 1,2-dichloro-4-nitrobenzene (DCNB), 1-chloro-2,4-dinitro benzene (CDNB), sulfobromophthalein (BSP) and benzalacetone (PBO)] were determined. There were no significant differences between young and old rats in the liver enzyme activities before the PFD. The PFD caused significant decreases in activities for three substrates (DCNB, BSP and STOX) in both age groups, with no significant differences between young and old rats a day 0. During recovery from the PFD, activities for the three substrates exceeded basal levels in young rats but at different time intervals (STOX, day 2; BSP, day 5; DCNB, day 9), while enzyme values in old rats tended to return slowly to basal values with no "overshoot." Concentrations of subunits 3 and 4 in young rat livers that were depressed by the PFD did not recover until day 9 of the ND, while subunits 2 and especially 1 increased during the ND refeeding, overshooting the basal levels. In contrast, in old rat livers the only change was a reduction of subunit 1 by the PFD and its gradual recovery during ND refeeding. These results demonstrate that our previous observation of overshooting of enzyme activities in mice is reproducible in rats but with certain substrate specificities. Furthermore, changes in subunit concentrations caused by aging and a PFD are more complex than what was predicted from changes in enzyme activities of GSTs.

Aging↗
PubMed1990

Difference in the effects of ursodeoxycholate and its taurine conjugate on the biliary excretion of sulfobromophthalein in the rat.

A simultaneous intravenous infusion of sulfobromophthalein (BSP) (0.3 mg.min-1.100 g-1) and ursodeoxycholate (UDC) or tauroursodeoxycholate (TUDC) (1.2 mumol.min-1.100 g-1) caused a significantly higher excretion rate of BSP than in the control value without bile salt infusion. In UDC-infused rats, however, the BSP excretion rate began to fall rapidly within the first 30 min of infusion, while the bile flow rate remained high, or even continued to increase. In contrast, the BSP excretion rate in TUDC-infused rats was stable up to 60 min and then began to decline in parallel with the fall of bile flow rate. A significant increase in the BSP transport maximum (Tm) induced by UDC and TUDC in rats does not agree with a previous observation in hamsters where neither of these bile salts enhanced BSP Tm. The discrepancy between these two studies can best be explained by the poor biliary excretion of either TUDC or UDC in hamsters as was previously reported. It was concluded that in rats both TUDC and UDC can significantly increase BSP Tm, which paralleled the excretion rate of bile salts and not the bile flow rate.

Animals↗
PubMed1990

Necessity of hyperglycemia for effects of endogenous cholecystokinin on insulin and pancreatic exocrine secretion in conscious rats.

The effects of endogenous cholecystokinin (CCK) on insulin and pancreatic exocrine secretion were examined in conscious rats. Rats with bile and pancreatic fistulae, one duodenal cannula, and two jugular vein cannulae were divided into four groups, with and without glucose infusion (0.2 g/(ml.h)), and with bile and pancreatic juice (BPJ) diversion and return. Without glucose infusion, BPJ diversion did not have any significant effect on the plasma level of insulin or glucose; but with glucose infusion, it caused a significant increase in plasma insulin concentration 1 h after the diversion. The plasma glucose concentrations in the groups with BPJ diversion and return were not significantly different, and the response of pancreatic exocrine secretion to BPJ diversion was suppressed slightly, but not significantly, by glucose infusion. The plasma CCK concentration was increased significantly by BPJ diversion, but not affected by glucose infusion. These results indicate that, as observed in in vitro experiments with exogenous CCK, in conscious rats, endogenous CCK stimulates both exocrine and endocrine secretion of the pancreas, and that hyperglycemia is necessary for these effects to become apparent.

Animals↗