• Toxicological Research
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• 제19권1호
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• pp.67-72
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• 2003

Cholestatic liver injury results from the accumulation of toxic bile salts within the liver. The aim of the present study is to elucidate the changes in expression and cellular localization of apoptosis related proteins in the liver of bile duct-ligated (BDL) rat. Extrahepatic cholestasis was induced by double ligation of the common bile duct and cut between the ligatures. Animals were sacrificed at day 3 and at week 1, 2, 4, 6, and 8 after BDL. The number of TUNEL positive cells was increased significantly after 3 days of BDL, decreased over 2 weeks and remained constant thereafter. Fas expression was not changed and activation of caspase 8 did not occur. Fas immunoreactivity was exclusively observed in the cytoplasm of hepatocytes, indicating that Fas expressed in rat hepatocytes is a soluble form. Hepatocyte apoptosis was associated with Bax expression, which showed a peak at day 3 and decreased over time gradually. Immnunostaining of Bax was observed in hepatocytes and bile duct epithelial cells (BEC) of control and BDL rats. Bcl-2 was increased over time in BDL rats. These results suggest that apoptosis of hepatocytes in BDL rats is independent of Fas and controlled by Bax expression.

• Applied Microscopy
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• 제24권2호
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• pp.78-92
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• 1994

Lysozyme has been reported to be present in the secretory granules of the Paneth cell, and lysozyme immunoreactivity has been detected by immunogold method in Paneth cells of the intestine of human, mouse and rat. The present study was aimed at clarifying the intracellular distribution and changes of the lysozyme immunoreactivity in rabbit Paneth cell after common bile duct ligation of rabbit, using the electron microscope immunogold technique. Healthy adult rabbits weighing about 2kg body weight were divided into normal and bile duct ligated groups. Common bile duct ligation was performed aseptically under ether anesthesia. Experimental animals were sacrificed on the 1st, the 3rd, the 5th, the 7th and the 14th day after the operation. Mucosal specimens from the intestinal gland of ileum were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde, followed by 1% osmium tetroxide, embedded in araldite mixture, cut with LKB-V ultratome. Ultrathin sections were placed on parlodion coated nickel grids (200mesh). The section-bearing grids were floated upside down on the added substance in a moist chamber at room temperature except for the primary antibody step, which was at $4^{\circ}C$. Sections were etched with a saturated solution of sodium m-periodate for 60min. After etching, sections were pretreated with 0.02M tris buffered saline (TBS), pH 8.4, with 1% bovine serum albumin (BSA, Sigma) for 60min, then treated polyclonal rabbit anti-human lysozyme (Dakipatts) diluted 1 : 50 in TBS with 0.1% BSA for 20hr. Subsequently, grids were incubated 60min in biotinylated goat anti rabbit IgG (Amersham) diluted 1 : 100 in TBS with 0.1% BSA. After this, sections were incubated 60min on streptavidin gold G10 (Amersham) diluted 1 : 50 in TBS with 0.1% BSA. After each step, the grids were briefly rinsed with TBS with 0.1% BSA. After the strepavidin gold step, the sections were jet washed with distilled water. Counterstain of the sections performed by uranyl acetate and lead citrate, and observed with JEM 100 CX II electron microscope. Observed results were as follow; 1. Secretory granules of mouse Paneth cells have a lysozyme immunoreactivity and also eosinophil leucocyte of rabbit applied for the positive-control stain, are well labeld with gold particles. 2. Normal rabbit Paneth cells have a lysozyme immunoreactivity restricted on the secretory granules. 3. Amount lysosomes containing myelin figures in the Paneth cells were significantly increased from 5th day after the common bile duct ligation. 4. Immunoreactivity of Paneth cell secretory granules were more activated on the 3rd day after the common bile duct ligation as compared with those of the normal animal. But the lysozyme immunoreactivity were decreased from the 5th day after the common bile duct ligation. 5. Considering the above finding, lysozyme contained Paneth cell are affected following of common bile duct ligation, whereas lysosomes containing myelin-figure do not exhibit any immunoreactive relationship with those of secretory granules.

• The Korean Journal of Physiology and Pharmacology
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• 제23권4호
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• pp.263-270
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• 2019

Hydrogen sulfide is well-known to exhibit anti-inflammatory and cytoprotective activities, and also has protective effects in the liver. This study aimed to examine the protective effect of hydrogen sulfide in rats with hepatic encephalopathy, which was induced by mild bile duct ligation. In this rat model, bile ducts were mildly ligated for 26 days. Rats were treated for the final 5 days with sodium hydrosulfide (NaHS). NaHS ($25{\mu}mol/kg$), 0.5% sodium carboxymethyl cellulose, or silymarin (100 mg/kg) was administered intraperitoneally once per day for 5 consecutive days. Mild bile duct ligation caused hepatotoxicity and inflammation in rats. Intraperitoneal NaHS administration reduced levels of aspartate aminotransferase and alanine aminotransferase, which are indicators of liver disease, compared to levels in the control mild bile duct ligation group. Levels of ammonia, a major causative factor of hepatic encephalopathy, were also significantly decreased. Malondialdehyde, myeloperoxidase, catalase, and tumor necrosis factor-${\alpha}$ levels were measured to confirm antioxidative and anti-inflammatory effects. N-Methyl-D-aspartic acid (NMDA) receptors with neurotoxic activity were assessed for subunit NMDA receptor subtype 2B. Based on these data, NaHS is suggested to exhibit hepatoprotective effects and guard against neurotoxicity through antioxidant and anti-inflammatory actions.

• 대한의생명과학회지
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• 제11권4호
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• pp.473-479
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• 2005

Possible mechanism of decreased catechol-O-methyltransferase (COMT) activity in cholestatic rat liver was studied. Hepatic and serum COMT activities were determined from the experimental rats with common bile duct ligation (CBDL). The Michaelis-Menten constants in this hepatic enzyme were also measured. The activities of cytosolic, mitochondrial and mircosomal COMT as well as their Vmax values were found to be decreased significantly in CBDL plus taurocholic acid (TCA) injected group than in the control group, such as CBDL alone groups. However, their Km values in the experimental groups did not vary. Serum COMT activity increased slightly in the CBDL plus TCA injected group than in the control group. The above results suggest that TCA represses biosynthesis of the COMT in the liver. The elevated activity of the serum COMT is believed to be caused by the increment of membrane permeability of hepatocytes upon TCA mediated liver cell necrosis.

• Archives of Pharmacal Research
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• 제23권5호
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• pp.501-506
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• 2000

We examined the antifibrotic effect of a methanol extract from Stephania tetrandra (ST) on experimental liver fibrosis. liver fibrosis was induced by bile duct ligation and scission (BDL/S) in rats. In BDL/S rats, activity levels of aspartate transaminase (AST), alanine transaminse (ALT), alkaline phosphatase (ALP), concentration of total bilirubin in serum, and hydroxyproline content of the liver were significantly increased. The ST treatment (either 100 ${m}g/kg/day$ or 200 ${m}g/kg/day$, p.o. for 4 weeks) in BDL/S rats reduced the serum AST, ALT and ALP activity levels significantly (p<0.01). Similarly, when compared to the control group, the concentration of hydroxyproline in the livers of the BDL/S rats treated with 100${m}g$ or 200${m}g$ ST treated rats decreased by 40% and 33% respectively, when compared to the BDL/S control group (p<0.01). The morphological characteristics of fibrotic liver that were observed in the BDL/S control group, improved in the ST treated BDL/S group. In the fibrotic liver of BDL/S rats treated with ST, a marked reduction in the numbers of alpha smooth muscle cell actin positive stellate cells was observed. These results indicate that doses of either 100 or 200 ${m}g/kg/day$ of methanol extract from S. tetrandra, had an antifibrotic effect in rats with liver fibrosis induced by bile duct ligation and scission.

• Advances in pediatric surgery
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• 제9권1호
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• pp.52-56
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• 2003

A 10 year old boy was admitted with blunt abdominal trauma by bike handle injury. The patient was operated upon for a generalized peritonitis due to pancreaticoduodenal injury. On opening the peritoneal cavity. complete transection of distal end of common bile duct and. partial separation between pancreas head and second portion of duodenum were found. Ligation of the transected end of the common bile duct. T-tube choledochostomy, and external drainage were performed. A pseudocyst was found around the head portion of the pancreas on the 7th postoperative day with CT. An internal fistula had developed between the pseudocyst and ligated common bile duct. The pseudocyst was subsided after percutaneous drainage. In the case of the undetermined pancreatic injury, percutaneous external drainage can be effective in treating the traumatic pancreatic pseudocyst in a pediatric patient.

• Applied Microscopy
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• 제22권2호
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• pp.15-29
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• 1992

The gallbladder is known to have the function of the storage and the concentration of the bile produced by the liver. This function is carried out by the removal of water and inorganic electrolytes. Extrahepatic cholestasis or the impairment of excretion of the bile leads to the distension and loss of the function of the gallbladder. The purpose of this study was to examine the ultrastructural characteristics of the normal gallbladder epithelial cells, and their structural changes induced by the ligation of common bile duct of the rabbit. Common bile duct ligation was performed under ether anesthesia. The rabbits were sacrificed on the 1st, 3rd, 5th, 7th and 14th day, respectively after operations. The tissue blocks of the gallbladder were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde prior to fixation in 1% osmium tetroxide, and embedded in the araldite mixture, and observed with JEM 100 CX-II electron microscope. The results were as follows: 1. The normal gallbladder epithelium of adult rabbit demonstrated two cell types, the ordinary epthelial cell and the dark cell. The dark cells have electron dense cytoplasm, and were found much infrequently, whereas ordinary epthelial cells were found quite numerous. 2. The ordinary epthelial cells of normal gallbladder were provided with the regular microvilli at the free surface and the images of pinocytotic activities in the apical cytoplasm, and exhibit highly convoluted lateral surfaces with elaborated microfolds. These figures of the cells suggest that they are resorptive in functional activity. 3. In the early stages (1st, 3rd, 5th day groups) following the ligation, the apical cytoplasm of some cells is protruding from the free surface and lost their microvilli. Numerous mucous granules filled in the apical and supranuclear cytoplasm compactly. 4. In the late stages (7th, 14th day groups) following the ligation, many light cells containing mumerous mucous granules are seen, between the ordinary epthelial cells. Mucous granules are fused each other, and are discharged into the lumen from the apical cytoplasm. The lateral membranes are straight or undulating without any interdigitations. From the above results, it was concluded that in the cholestasis induced by the common bile duct ligation, there is a tendency for the mucosal epithelium of the rabbit gallbladder to have secretory rather than an absorptive function.

• Applied Microscopy
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• 제27권3호
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• pp.295-308
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• 1997

본 연구에서는 dehydrocholic acid와 biligrafin 투여 20분 후, 기아 8일 후 및 담관결찰 48시간 후의 흰쥐 간을 동결할단하여 담세관 주위 폐쇠띠의 미세구조를 관찰하였다. 정상대조군에서 폐쇠띠는 여러부위에서 $1\sim4$개의 줄로 이루어진 망상구조를 보였다. dehydrocholic acid 투여군에서는 폐쇠띠의 폭이 넓어져서 망상구조의 줄은 $6\sim9$개로 증가된 부위가 많이 나타났다. 그러나 biligrafin 투여군에서는 줄이 감축되어 있었으며 망상구조는 흩으러져 있었다. 기아군에서는 많은 부위에서 폐쇠띠가 $2\sim3$개의 줄로 이루어져 있었으며 이들 줄은 느슨하게 연결되어 있었다. 결찰군에서도 많은 부위에서 폐쇠띠의 망상구조는 흩으러져 있었으며 줄들이 감축되어 있거나 아직 감축되지 않은 상태로 있는 부위도 관찰되었다. 그러나 모든군에서 줄들은 틈을 보이면서 단절되어 나타나거나 선상으로 배열된 입자로 이루어져 있었는데 특히 biligrafin군과 결찰군에서 이러한 소견이 현저하였다. 줄의 끝이 유리되어 있는 것들은 기아군에서 많이 관찰되었는데 dehydrocholic acid군에서는 줄의 끝이 교통반에 접해 있거나 그 주위를 둘러싸고 있는 것들을 많이 볼 수 있었다. 줄들이 불연속적으로 나타나거나 끝이 유리된 것은 정상군에서도 드물지 않게 관찰되었다. 결찰군을 제외한 모든 군에서 담세관 주위 폐쇠띠가 그망안에 입자의 집괴를 함유하고 있었다. 이상의 증거로 미루어 담세관 주위의 폐쇠띠는 담세판의 상태에 따라 쉽게 변하는 것 같이 보이며 같은 담세관에서 일지라도 부위에 따라 구조적 변화에 차이가 있는데 이러한 소견은 정상상태하에서도 비슷하게 나타나고 있어 폐쇠띠가 완전한 장벽으로서의 역할을 하고 있지 않는 경우도 있는 것 같이 보인다. 또한 폐쇠띠는 기계적 자극이나 다른 어떠한 인자에 의해서 쉽게 영향을 받는 것 같이 보이며 폐쇠띠의 변화에 연관하여 줄과 입자의 집괴 사이에도 어떠한 관계가 있을 것으로 추정된다.

• 대한수의학회지
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• 제23권1호
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• pp.25-35
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• 1983

In order to assess the diagnostic aid of serum gammaglutamyl transpeptidase values in hepatitis, obstructive jaundice and pancreatitis, four groups of 14 health dogs were subjected to the gastric intubatin of $CCl_4$(1.5ml/kg of body weight), the ligation of common bile duct, the ligation of pancreatic ducts and the injection of chloroform(0.2ml/kg of body weight) in the parenchyma of the pancreas. Some serum enzymes serum glutamic pyruvic transaminase(SGPT), serum glutamic oxalacetic transaminase(SGOT), total bilirubin, amylase and lipase known to be indicative of hepatic and pancreatic diseases were monitored. In comparision of these enzymes, gamma-glutamyl transpeptidase(GGTP) valuers were determined in these dogs before and after the experimental procedures. The results were summarized as follows: 1. In $CCl_4$ intoxication gorup, there were no significant changes in serum GGTP activities(mean: 6.0~14.6 IU/L). 2. In bile duct ligation group, serum GGTP activities shelved marked increases, beginning at postsurgical day 1 and rose the highest mean value(342.7 IU/L) on day 12. Then the activities never approached to the base-line values. 3. After the ligation of pancreatic ducts and the injection of chloroform in the pancreas, serum GGTP activities did not rise throughout the experiment. 4. SGPT:GGTP ratio did not increase in bile duct ligation group, but increase markedly in $ccl_4$ intoxication group. 5. The results indicated that serum GGTP values or SGPT:GGTP ratio could provide valuable indicators for differential diagnosis between hepatobiliary obstruction and hepatocellular disease.

• BMB Reports
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• 제32권4호
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• pp.331-338
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• 1999

Ethanol catabolism is thought to produce metabolic disorders resulting in alcoholic liver disease. To investigate the mutual effects of ethanol catabolism and cholestasis induced by common bile duct ligation on the activities of carboxylesterase, we have determined the enzyme activities in rat hepatic (cytosolic, mitochondrial, and microsomal) preparations as well as in rat serum using ten animal models: normal rats (group 1), sham-operated rats (group 2), common bile duct-ligated rats (group 3), ethanol-intoxicated rats (group 4), sham-operation plus chronic ethanol-intoxicated rats (group 5), common bile duct-ligated plus chronic ethanol-intoxicated rats at 1.5h and 24h (groups 7A and 7B), and duct-ligated and acute ethanol intoxicated rats at 1.5 h and 24 h (groups 8A and 8B). The $K_m$ and $V_{max}$ values of carboxylesterase from these hepatic preparations of cholestatic rat liver combined with chronic ethanol intoxication were also measured by using ethyl valerate as the substrate from the 14th day post-ligation. Carboxylesterase activities of all hepatic preparations and rat serum (group 3) showed significant decreases compared to the activities from the sham-operated control (group 2). Enzyme kinetic parameters indicated that $V_{max}$ of carboxylesterase from all the hepatic preparations in cholestatic rats (group 3) decreased significantly, although the $K_m$ values were about the same as in the sham-operated control (group 2). When cholestasis was combined with chronic ethanol intoxication (group 6), carboxylesterase activities showed further decrease in all the hepatic preparations and serum compared to the control activity (group 5). The $V_{max}$ also decreased significantly, although $K_m$ values did not change. When common bile duct ligation was combined with acute ethanol intoxication (group 8), the enzyme activities in the rat liver and serum showed significant decrease compared to the activity from acute ethanol-intoxicated rats (group 7). However, quite contrary to this, the activities of serum from acute ethanol intoxication 1.5 h (group 7A) increased significantly compared to the activities in the normal control (group 1). These results, therefore, suggest that the biosynthesis of hepatic carboxyl-esterase seems to decrease when cholestasis is combined with chronic and acute ethanol intoxication, and the decrease in activity is more significant than from cholestasis alone.