• Applied Microscopy
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• v.28 no.1
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• pp.83-90
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• 1998

The pathway of intracellular transport of dehydrocholic acid was investigated in the hepatocytes of rats by transmission electron microscopy with conventional and freeze fracture methods. Both in normal and experimental groups, the cis Golgi cisterns were sacculated and faces toward the bile canaliculus. In the experimental group, however, the cis Golgi cisterns showed buds, which were probably separated to be vesicles. Some of the buds were connected to the cisterns with the narrow neck. The vesicles were increased in the vicinity of bile canaliculi. The fusion between vesicles and bile canaliculus were frequently observed in the experimental group. This was particularly well shown in the freeze fracture replica. In the thin section, the vesicles were devoid of visible contents as seen in the bile canaliculli. The evidence suggests that the vesicles are derived from the cis Gogi cistern in the way that buds pinch off, serve as vehicles to transport dehydrocholic acids and fuse to bile canaliculi for exocytosis.

• Applied Microscopy
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• v.26 no.4
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• pp.465-477
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• 1996

The influence of dehydrocholic acid, cholesterol and phosphstidylcholie to the fine structure of vacuolar apparatus was investigated to better understand the mechanism of intracellular transport of bile constituents in the hepatocytes of rats. The cis Golgi cisterns faced toward the bile canaliculi both in normal and experimental groups. In the hepatocytes from the rats of experimental groups, the primary organic solutes in bile influence the Gogi apparatus, ER and lysosome in the way of increase, cisternal dilation or budding to form the vacuoles. In the dehydrocholic acid group, the cis Golgi cisterns appeared to be sacculated and showed buds, which were probably separated to be vacuoles. Some of the vacuoles appeared to be fused to the bile canaliculi. In the cholesterol and phosphatidylcholine groups, the Golgi cisterns appeared to be dilated and lysosomes were increased in the vicinity of bile canaliculi. The cis Golgi cisterns showing linear saccular fashions were occasonally observed. The increase of lysosomes were more predominant in the cholesterol group. The evidence suggests that dehydrocholic acid is mainly transported through the ER and cis Golgi cisterns, and cholesterol and phosphatidylcholine are mainly transported through the ER and lysosomes via the trans Golgi cisterns, but the cholesterols are frequently transported via the lysosomes.

• Applied Microscopy
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• v.27 no.3
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• pp.295-308
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• 1997

Liver tissues were taken from rats 20 min after administration of dehydrocholic acid and biligrafin, fasted for 8 days, and bile duct ligation for 48 hr. And the zonulae occludentes around the bile canaliculi were investigated by the freeze fracture method. In the normal rats, the zonulae occludentes showed networks consisting of one to four strands in many regions. After administration of dehydrocholic acid, the depth of zonulae occludentes were increased and networks consist of six to nine strands. The zonulae occludentes, however, were disorganized, and strands were reduced after administration of biligrafin. In the fasting rats, the zonulae occludentes showed one or two strands which were loosely linked. After ligation of bile duct, the zonulae occludentes were disorganized, and strands were reduced or still in a state not to be reduced. The strands were interrupted by gaps or linear particles in all the groups, especially in the biligrafin and the ligation. The free ends were remarkable in the fasting group and contact or surround the lateral gap junctions in the dehydrocholic acid group. The discontinuities and free ends of strands were also encountered in the normal group, thus suggesting that the barrier is not complete even in the normal state. The aggregation of particles appeared within the meshwork of zonulae occludentes in all the group, except for the ligation. From the evidence, it is assumed that zonulae occludentes are labile, differ in the structure according to the situation of bile canaliculi and show regional alterations even in the same bile canaliculi, that zonulae occludentes are affected by mechanical stress or certain other factors, and that there is a correlation between the strands and particle aggregations in association with the alteration of zonulae occludentes.

• Applied Microscopy
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• v.28 no.2
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• pp.171-180
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• 1998

In the present study, the vacuolar apparatus were investigated in the hepatocytes of rats treated with DA by transmission electron microscopy of conventional and cytochemical thin sections. In the rats after 20 min of dehydrocholic acid treatment, the cis Golgj cisterns were sacculated in line. The saccule occasionally occured by elongation and attenuated neck. The lysosomes also showed protrudent saccule. The vesicles were observed near the cis Golgi cisterns, lysosome and bile canaliculi. Some of the vesicles appeared to be fused to bile canaliculi. The cis Golgi cisterns usually faced toward the bile canaliculi both in normal and experimental groups. The cis Golgi cisterns, protrudent saccule and vesicles were almost devoid of visible contents. The osmium deposits were heavy on the protrudent saccule as well as on the cis Golgi cisterns or on the vesicles isolated near by, but they were light or not observed on the vesicles in the immediate vicinity of bile canaliculi. The acid phosphatase activities appeared on the lysosome and vesicles located near by, but did not appear on the vesicles as approaching closer to the bile canaliculi. The evidence suggests that the vesicles are derived from the cis Gogi cistern and lysosomes and fuse to bile canaliculi for exocytosis, and that the activity in the vesicles is diminished as approaching closer to the bile canaliculi.

• Biomolecules & Therapeutics
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• v.4 no.2
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• pp.162-166
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• 1996

This study was done to investigate whether cholic acid derivatives have anti-stress activity and what is a cause of this anti-stress effect. Seven cholic acid derivatives (cholic acid, taurocholic acid, ursodeoxycholic acid, tauroursodeoxychoic acid, chenodeoxy cholic acid, dehydrocholic acid, hyodeoxycholic acid) were used, silymarin and valproic acid were used as positive controls. Stress was induced by restraint immobilization technique plus water immersion (24hrs) and adrenal weight, spleen weight, adrenal ascorbic acid, serum cholesterol, lactate dehydrogenase (LDH), alkaline phosphatase (ALP), adrenal cholesterol, glucose and corticosterone levels were measured as stress indicators. Most cholic acid derivatives markedly decreased the adrenal weight, and TUDCA and DHCA increased the spleen weight. The restraint stress induced increments in serum LDH, ALP and cholesterol were attenuated by most cholic acid derivatives. Cholic acid, taurocholic acid and tauroursodeoxycholic acid only increased the content of adrenal ascorbate. While valproic acid showed an inhibitory effect against stress, silymarin did not. Our findings suggest that most cholic acid derivatives have anti-stress effect and that their anti-stress effect is, in part, related to choleretic activity.

• The Korean Journal of Pharmacology
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• v.18 no.1
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• pp.43-54
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• 1982

Bile formation is a complex process comprised of three separate physiologic mechanism operating at two anatomical sites. At present time, it was known that at least two processes are responsible for total canalicular secretion at the bile canaliculus. One of the processes is bile salt-dependent secretion (BSDS) hypothesis that the active transport of bile salts from plasma to bile provided a primary stimulus for bile formation: the osmotic effect of actively transported bile acid was responsible for the movement of water and ions into bile. The other process is bile salt-independent secretion (ESIS), which is unrelated to bile salt secretion at the canaliculus and which may involve the active transport of sodium. The third process for bile formation involves the biliary ductal epithelium. Secretin-stimulated bile characteristically contained bicarbonate in high concentration. Therefor, it was suggested that secretin stimulated water and bicarbonate secretion from the biliary ductules. One the other hand, it was found that a large amounts of cAMP was present in canine bile but no apparent relationship between bile salt secretion and cAMP content in dog bile. However, bile flow studies in human have demonstrated that secretin and glucagon increase bile cAMP secretion as does secretin in baboons. Secretin increases baboon bile duct mucosal cAMP levels in addition to bile CAMP levels suggesting that in that species secretin-stimulated bile flow may be cAMP mediated. It has been postulated that glucagon and theophylline which increase the bile salt-independent secretion in dogs might act through an increased in liver cAMP content. In a few studies, the possible role of cAMP on bile formation has teen tested by administration of an exogenous derivative of cAMP, dibutyryl cAMP. In the rat, DB cAMP did not modify bile flow, but injection of DB cAMP in the dog promoted an increase in the bile salt-independent secretion. Because of these contradictory results, this study was carried out to examine the relationship between cyclic nucleotides and bile flow due to various bile salts as well as secretin or theophylline. Experiments were performed in rabbits with anesthesia produced by the injection of seconal(30 mg/kg). Rabbits had the cystic duct ligated and the proximal end of the divided common duct cannulated with an appropriately sized polyethylene catheter. A similar catheter was placed into the inferior vena cava for administration of drugs. Bile was collected for determination of cyclic nucleotides and total cholate in 15 min. intervals for a few hours. The results are summerized as followings. 1) Administrations of taurocholic acid or chenodeoxycholic acid increased significantly the concentrations of cAMP and cGMP in bile of rabbits. 2) Concentration of cAMP in bile during the continuous infusion of ursodeoxycholic acid, was remarkedly increased in accordance with the increase of bile flow, while on the contrary concentration of cGMP in bile was decreased significantly. 3) Dehydrocholic acid and deoxycholic acid significantly increased bile flow, total cholate output and cyclic nucleotides in bile. 4) Only cAMP concentration in bile was significantly increased from control value by secretin, while theophylline increased cAMP as well as cGMP in rabbit bile. 5) In addition, the administration of secretin to taurocholic acid-stimulated bile flow increased cAMP while theophylline produced the increases of cAMP and cGMP in bile. 6) The administration of insulin to taurocholic acid-stimulated bile flow decreased cAMP concentration, while on the contrary cGMP was remarkedly increased in rabbit bile.