• Archives of Pharmacal Research
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• v.27 no.4
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• pp.422-428
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• 2004

Expressed sequence tags (ESTs) were generated from two 3'-directed CDNA libraries constructed from quiescent and activated rat hepatic stellate cell (HSC) to analyze the expression profiles of active genes in both cells. From quiescent and activated HSC, 694 ESTs and 779 ESTs, respectively, were obtained after excluding those having shorter than 30 bp. Amonq ESTs obtained from quiescent and activated HSC, 68 and 73 kinds of ESTs (186 clones and 236 clones), respectively, appeared more than once, implying that their genes are expressed highly in each cell type. 52 among 73 ESTs appeared only in the activated HSC 47 amonq 68 ESTs only in the normal HSC, and 21 in both cells. The genes of these 52 ESTs were assumed to be expressed more highly in the activated HSC. To confirm the high expression of genes of which the ESTs appeared more than twice in the activated HSC, northern hybridization was carried out with RNAs derived from rat normal and fibrotic liver using each of 18 EST DNAs as probe. 13 ESTs showed more intense bands with RNA isolated from the fibrotic liver than normal liver. From these results, we confirm the positive correlation between abundance of transcript in activated HSCs and the expression level in fibrotic liver, The expression profile of the transcripts serves as an important tool in understanding the biological properties of HSC.

• Archives of Pharmacal Research
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• v.20 no.5
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• pp.459-464
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• 1997

The human liver cDNA clone UDPGTh2, encoding a liver UDP-glucuronosyltransferase (UDPGT) was isolated from a .gamma. gt 11 cDNA library by hybridization to mouse transferase cDNA clone, UDPGTm1. UDPGTh2 encoded a 529 amino acid protein with an amino terminus membrane-insertion signal peptide and a carboxyl terminus membrane-spanning region. There were three potential asparagine-linked glycosylation sites at residues 67, 68, and 315. In order to obtain UDPGTh2 protein encoded from cloned human liver UDP-glucuronosyltransferase cDNA, the clone was inserted into the pSVL vector (pUDPGTh2) and expressed in COS 1 cells. The presence of a transferase with Mr-52,000 in transfected cells cultured in the presence of $[^{35}S]$ methionine was shown by immunocomplexed products with goat antimouse transferase IgG and protein A-Sepharose and analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. The expressed UDPGT was a glycoprotein as indicated by electrophoretic mobility shift in Mr-3,000-4,000 when expressed in the presence of tunicamycin. The extent of glycosylation was difficult to assess, although one could assume that glycosyl structures incorporated at the level of endoplasmic reticulum were always the core oligosaccharides. Thus, it is likely that at least two moieties inserted can account for the shift of Mr-3,000-4,000. This study demonstrates the cDNA and deduced amino acid sequence of human liver UDP-glucuronosyltransferase cDNA, UDPGTh2.

• Applied Microscopy
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• v.18 no.2
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• pp.205-217
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• 1988

An ultrastructural study of hepatocyte proliferation in the regenerating rat liver has been made by means of the partial hepatectomy. And electron microscopic histochemistry of hepatocyte in the regenerating rat liver is studied through alkaline phosphatase reaction. The results are as follows: 1. When the regeneration of rat liver is induced by the partial hepatectomy, the prominent ultrastructural characteristics of hepatocyte are changes of the distribution of chromatin in nucleus, increase of the number of mitochondria and decrease of the size of them, development of rough endoplasmic reticulum, and transient decrease of glycogen granules in cytoplasm. 2. Alkaline phosphatase reaction products are appeared in the nucleus or rough endoplasmic reticulum of hepatocyte during the initial regeneration of liver as 24, 48 and 72 hour groups after partial hepatectomy. And these positive reaction are mainly increased in cytoplasm and plasma membrane of hepatocytes during 1, 2 and 3 week groups after partial hepatectomy. As 4 weeks passed after partial hepatectomy, these positive reaction is located in the sinusoidal epithelial cells or erythrocytes. With above results, we concluded that alkaline phosphatase was synthesized in the rough endoplasmic reticulum bounded ribosomes of regenerating hepatocyte, was transported to the plasma membrane of them, and then was transported in blood by the way sinusoidel epithelial cells.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.305-306
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• 2003

To treat fulminant hepatic failure (FHF) patients, various extracorporeal bioartificial liver (BAL) systems have been developed. Several requirements should be met for the development of BAL systems: hepatocytes should be cultured in a sufficiently high density; their metabolic functions should be of a sufficiently high level and duration; and the BAL systems module should permit scaling-up and aseptic handling. Several investigators have found that freshly isolated primary hepatocytes can be cultured into three dimensional, tightly packed, freely suspended, multicellular aggregates, or spheroids. These specialized cell structures exhibited enhanced liver specific functions and a prolonged differentiated state compared to cells maintained in a monolayer culture. Cells in spheroids appear to mimic the morphology and ultrastructure of the in vivo liver lobule. The ability of hepatocytes to organize into three-dimensional structures was hypothesized to contribute to their enhanced liver-specific activities. In this study, the ammonia removal rate and urea secretion rate of pig hepatocytes spheroids encapsulated in Ca-alginate bead were determined. A packed-bed bioreactor with encapsulated pig hepatocytes was devised as BAL support system. The efficacy of the system was evaluated in vitro.

• The Korean Journal of Pharmacology
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• v.7 no.1
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• pp.37-52
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• 1971

It has been demonstrated repeatedly that the administration of ethionine, a methionine analogue, will produce destruction of pancreatic tissue and liver cells. Destruction of acinar cells of pancreas by the administration of excess methionine similar to that seen after feeding diets supplemented with ethionine was also reported, but the liver was not involved by this amino acid. In an attempt to reproduce the results of these investigators the tissue damages were found to be slight and seen only irregulary in rats receiving ethionine on a complete diet and the result of tissue damages by excess methionine were also controversial. The present studies describe the toxicity of dietary supplemented ethionine and methionine to liver and pancreas in rats fed a low protein diet. Hundred five albino rats weighing around 120 gm were divided into three groups as follows; 1) Control group: A low protein diet containing 8% casein was fed throughout the experimental period. 2) Methionine group: A low protein diet (7% casein) supplemented with 1% methionine was used. 3) Ethionine group: A similar diet as methionine group except the supplementation of 1% ethionine instead of methionine was used. Five animals per wee from each group were killed for 6 weeks. The liver and pancreas were fixed in 10% formalin and histologic sections were prepared and stained with hematoxylin eosin. Serum amylase was expressed as much of glucose liberated from a starch substrate. The glucose was determined by the method of Nelson (1944). Serum glutamic pyruvic transaminase (SGPT) and glutamic oxaloacetic transaminase (SGOT) were determined by the technique described in Sigma Bulletin.

• Korean Journal of Veterinary Service
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• v.25 no.4
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• pp.357-370
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• 2002

This experiment was performed to investigate apoptosis during undergoing patho-genesis of Salmonella gallinarum(SG)-infected chicks. 16 days old, 49 chicks was infected with SG (10$\^$6/-10$\^$8/ CFU/㎖) experimentally, they were autopsied to remove liver, spleen, intestine and lung at 1, 6, 12hr, 1, 2, 4 and 7 day post infection(PI) respectively, for H-E and TUNEL staining. Grossly, white foci in the liver and enlarged spleen were seen on 4 day PI and coppery bronze liver, dark-red discolored intestine, green-yellowish discolored and enlarged spleen was observed on 7 day PI. Histopathologically, multi focal necrosis in the liver, follicle hyperplasia in the spleen and inflammatory cells infiltration in the intestine were shown from 2 day PI and more severely observed on 4 day and 7 day PI. In TUNEL analysis, apoptotic cells reached a maximum at 6hr PI in the liver and intestine and at 12hr PI in the spleen, and then decreased the levels of controls by 7 day PI.

• The Journal of Korean Medicine
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• v.32 no.1
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• pp.151-163
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• 2011

Objective: The purpose of this study is to observe the effects of Persicae Semen herbal-acupuncture at GB34(Yangneungcheon) on hyperlipidemia of rats caused by high-fat diet. Methods: The author performed several experiments to measure the DPPH radical scavenging activity of PS-HAS,to analyze various components and enzymes in serum and liver, and to observe the histological changes of liver and aorta. Results: 1. Persicae Semen herbal-acupuncture solution increased the DPPH radical scavenging activity in rat's liver cells. 2. In the Persicae Semen herbal-acupuncture group, compared with control group, the level of total cholesterol in serum and the ratio of total cholesterol to HDL-cholesterol decreased significantly. 3. In the Persicae Semen herbal-acupuncture group, compared with the control group, the level of glutathione in liver cells increased significantly. 4. In the Persicae Semen herbal-acupuncture group, compared with the control group, the pathological changes in liver and aorta were reduced. Conclusion: The above results suggest that Persicae Semen herbal-acupuncture at GB34 has good effects on preventing and treating hyperlipidemia.

• Natural Product Sciences
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• v.18 no.2
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• pp.76-82
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• 2012

Stamens of Mesua ferrea L. are a well-known herbal drug used in Indian System of Traditional Medicine to treat various diseases. The claimed activity of this plant part is necessitated to investigate antioxidant and hepatoprotective activity. Authenticated plant sample was extracted with hexane, ethanol (EtOH) and water (aq.) using ASE 100 accelerated solvent extractor. Antioxidant activity was evaluated by means of different in vitro assays. Hepatoprotective effect was investigated on carbon tetrachloride induced oxidative stress in liver slice culture model. Cytotoxic marker lactate dehydrogenase (LDH) released in culture medium and the activity of lipid peroxidation along with antioxidant enzymes (AOEs) namely superoxide dismutase (SOD), catalase (CAT) and glutathione reductase (GR) were estimated. Hexane and EtOH extracts were significantly inhibited DPPH, NO, SOD and $ABTS^+$ radical in dose dependent manner. The trade of phenol content was: aq. extract < hexane extract < EtOH extract. A significant correlation was shown by total phenol content and free radical scavenging activity of extracts. The culture system treated with hexane extract, EtOH extract or ascorbic acid exhibited significant depletion in LDH, lipid peroxidation, antioxidative enzymes SOD, CAT and GR. Hexane extract and EtOH extracts of stamen of M. ferrea protected liver slice culture cells by alleviating oxidative stress induced damage to liver cells.

• YAKHAK HOEJI
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• v.59 no.5
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• pp.201-206
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• 2015

The aim of the present study was to optimize in vitro method for the prediction of drug-induced liver injury using human liver microsomes (HLM). Cytotoxicity test of cyclophosphamide and acetaminophen in HepG2 cells cultured with HLM showed that the newly established condition using 0.375 mg/ml HLM for 24 hr incubation was comparable or more sensitive than the previously established condition using 0.75 mg/ml HLM for 12 hr incubation. Although the cytotoxic effect of troglitazone was completely attenuated by 0.75 mg/ml HLM, it was augmented by 0.375 mg/ml HLM in the presence of the NADPH-generating system. The cytotoxic effect of chlormezanone, a withdrawn drug due to hepatotoxicity in human, was increased by HLM in the presence of the NADPH-generating system. In contrast, the cytotoxic effect of methapyrilene, a withdrawn drug due to hepatotoxicity in rats, was decreased by HLM in the presence of the NADPH-generating system. The present study suggests that the optimized in vitro method using HLM can be useful for the prediction of drug-induced hepatotoxicity.

• The Journal of Internal Korean Medicine
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• v.26 no.4
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• pp.853-863
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• 2005

Object : This study was performed to investigate the anti-fibrogenic effect of Artemisiae Capillaris Herba(ACH) on cultured rat hepatic stellate cells. Methods : Hepatic Stellate Cells were obtained from a 350gm Sprague-Dawley rat by tissue perfusion system, and the cells for the study were selected after 3 passages of culture on non-coated plastic culture dishes which enable the cells to activate, thus producing collagens in the cell media. Cells were treated with various concentrations of Artemisia Capillaris Herba(ACH) extract powder for 24 or 48 hours. After the treatment, Soluble collagen, procollagen levels and the mRNA of the procollagen type I C were measured by using assay kit and RT-PCR method. Results : Procollagen production by the hepatic stellate cells decreased after the treatment in a time-dependent dose-dependent manner. The mRNA expression decreased consistently with the volume of the secreted procollagen which indicates the herb hat inhibitory effect on fibrogenesis of the liver by regulating one of the fibrosis associated genes in transcription. Conclusion : These results suggest that ACH is beneficial in the treatment of cirrhotic patients as well as for the patients with chronic hepatitis.