• Title/Summary/Keyword: Primary hepatocyte

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Effects of Monosodium Glutamate on Unscheduled DNA Synthesis and DNA Single-Strand Breaks in Primary Cultures of Rat Hepatocytes (일차배양 간세포에서 Monosodium Glutamate에 의한 돌연변이 유발성의 검증)

  • 김동현;양규환
    • Environmental Mutagens and Carcinogens
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    • v.7 no.2
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    • pp.59-64
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    • 1987
  • Cytotoxic and genotoxic potential of monosodium glutamate (MSG) were evaluated in primary cultures of rat hepatocytes. When exposed to liver cell culture continuously for 24 hr, MSG did not show any cytotoxic effects up to 0.5% (w/v) level as determined by Tryphan Blue exclusion and lactic dehydrogenase release test. MSG also did not induce unscheduled DNA synthesis or DNA single-strand breaks in hepatocyte cultures up to 1% level. No synergistic effects of MSG were observed on aflatoxin B$_1$-induced DNA damage when 1% MSG was treated to liver cell culture along with aflatoxin B$_1$.

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Effects of Aluminium on Vitellogenin and Its mRNA Induction by Estradiol-17$\beta$ in the Primary Culture of Hepatocytes in the Rainbow Trout Oncorhynchus mykiss

  • Hwang, Un-Gl;Park, Jin-Il;Shim, Jung-Min;Jung, Chang-Soo;Park, Sung-Yoon
    • Proceedings of the Korean Environmental Sciences Society Conference
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    • 2003.11a
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    • pp.159-164
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    • 2003
  • Effects of Al on vitellogenin (VTG) and VTG mRNA induction by estradiol-17 $\beta$($E_2$) were examined in primary hepatocyte culture of rainbow trout. Hepatocytes were precultured for 2 days and then E2 ($2{\times}10^{-6}$M) and Al ($10^{-6}-10^{-4}$M) were simultaneously added to the incubation medium. Hepatocytes were cultured for 5 more days. Media and hepatocytes were then analyzed by SDS-PAGE and Northern blotting for VTG and VTG mRNA, respectively. These metal had no appreciable effect on the viability of hepatocytes in culture. However, Al interfered with VTG production and VTG mRNA expression. Al reduced VTG production in a concentration-dependent way and a significant reduction accurred at Al concentrations greater than $5{\times}10^{-5}$M. VTG mRNA expression also decreased with a negative correlation with Al concentration (r=-0.98). These results suggest that Al inhibit VTG production at the transcriptional level to reduce VTG mRNA expression.

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Cytoprotective Effects of Natural Flavonoids on Carbon Tetrachloride-Induced Toxicity in Primary Cultures of Rat Hepatocytes (사염화탄소로 유도한 일차 배양 간세포 독성에서 Flavonoid류의 세포보호 효과)

  • Kim, Young-Kwan;Kim, Yang-Hee;Kim, Dong-Hyun;Lee, Kyung-Tae
    • Korean Journal of Pharmacognosy
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    • v.36 no.3 s.142
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    • pp.224-228
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    • 2005
  • Protective effects of various natural flavonoids on carbon tetrachloride $(CCl_4)-induced$ hepatotoxicity were investigated in primary cultured rat hepatocytes. Some of these flavonoids decreased the ALT and LDH releases induced by $CCl_4$ in A dose-dependent manner. Neohesperidin, hesperetin, baicalin, baicalein and quercetin inhibited $CCl_4-induced$ alanine aminotransferase (ALT) release. In addition, quercetin, quercitrin, neohesperidin, baicalin, baicalein and naringin reduced $CCl_4$ induced lactate dehydrogenase (LDH) leakage. Among these flavonoids, quercitrin, quercetin, baicalin and baicalein possessed potent protective effects and were selected for the further investigation on lipid peroxidation. These four flavonoids inhibited dose dependently $CCl_4-induced$ lipid peroxidation. Especially, the protective effects of quercetin and baicalein were similar to silybin as a well-known hepatoprotective agent. These results suggest that these four flavonoids have significant cytoprotective effects and possibility of therapeutic effect on chemical-induced liver diseases.

Hepatoprotective Activities of Gomisin A and Gomisin N (Gomisin A 및 Gomisin N의 간독성 보호작용)

  • Heo Jeong-Haing;Park Jin-Gu;Cheon Ho-Jun;Kim Yeong-Shik;Kang Sam-Sik;Hung Tran Manh;Bae Ki-Hwan;Lee Sun-Mee
    • Korean Journal of Pharmacognosy
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    • v.37 no.4 s.147
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    • pp.294-301
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    • 2006
  • The aim of this study was to investigate the protective activity of gomisin A and gomisin N, bioactive lignan components isolated from Schizandae Fructus, on hepatocyte injury induced by carbon tetrachloride($CCl_4$, 10 mM), t-butyl hydroperoxide(TBH, 0.5 mM), and D-galactosamine(GalN, 30 mM). Primary cultures of rat hepatocyte(18 h culture) were treated with $CCl_4$, TBH or GalN and various concentrations(0.1, 1, 10, $100{\mu}M$) of gomisin A or gomisin N. $CCl_4$ significantly increased the levels of lactate dehydrogenase(LDH), alanine aminotransferase(ALT), and aspartate aminotransferase(AST). These increases were inhibited by gomisin N. TBH significantly increased the level of AST; an increase that was inhibited by gomisin N. GalN markedly increased the levels of LDH and ALT, and these increases was significantly inhibited by both gomisin A and gomisin N. These results suggest that gomisin A and gomisin N have the hepatoprotective activity.

Hepatoprotective Activities of Curcumin, Demethoxycurcumin and Bisdemethoxycurcumin (Curcumin, demethoxycurcumin 및 bisdemethoxycurcumin의 간보호 작용)

  • Cheon, Ho-Jun;Park, Jin-Goo;Kim, Yeong-Shik;Kang, Sam-Sik;Chi, Xing-Fu;Lee, Jung-Joon;Lee, Sun-Mee
    • Korean Journal of Pharmacognosy
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    • v.38 no.2 s.149
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    • pp.139-147
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    • 2007
  • The aim of this study was to investigate the protective activity of curcumin, demethoxycurcumin and bisdemethoxycurcumin, isolated from Curcuma longa Linne, on hepatocyte injury induced by carbon tetrachloride (CCl$_4$,10 mM), t-butyl hydroperoxide (TBH, 0.5 mM) and D-galactosamine (GaIN,30 mM). Primary cultures of rat hepatocyte (18 h culture) were treated with CCl$_4$, TBH or GaIN and various concentrations (0.1, 1, 10 and 100 ${\mu}$M) of curcumin, demethoxycurcumin and bisdemethoxycurcumin CCl$_4$ significantly increased the levels of lactate dehydrogenase (LDH), alanine aminotransferase (ALT) and aspartate aminotransferase (AST). The increases in LDH, ALT and AST levels were inhibited by curcumin. Demethoxycurcumin and bisdemethoxycurcumin decreased the levels of AST. Curcumin inhibited the increases in ALT and AST levels induced by TBH. The increased levels of LDH, ALT and AST induced by TBH were inhibited by bisdemethoxycurcumin. GaIN markedly increased the levels of LDH, ALT and AST. These increases were significantly inhibited by bisdemethoxycurcumin. The increase in AST level was inhibited by curcumin. These results suggest that curcumin and bisdemethoxycurcumin have potent hepatoprotective activities.

Transition of Marker Enzymes of Rat Hepatocyte Organelles in Culture (배양중 흰쥐 간세포의 새포소기관 표지효소의 변천)

  • Song, In-Hwan;Kim, Joo-Yung;Sung, Eon-Ki;Lee, Yung-Chang
    • Journal of Yeungnam Medical Science
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    • v.6 no.2
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    • pp.133-140
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    • 1989
  • To investigate recovery, growth, and activity of hepatocyte in primary culture after cell separation, the authors followed up the marker enzyme activities of golgi complex, mitochondria and biologic membrane. Thiamine pyrophosphatase, the marker enzyme of golgi complex, activity approached the level of long term culture at 4th day. Succinate dehydrogenase, the marker enzyme of mitochondria, activity decreased with time, then it maintained constant level after 4th day. Alkaline phosphatase, the marker enzyme of biological membrane, activity increased from 3rd day, and after 5th day it showed strong reaction. These data suggested that hepatocytes were stabilized and recovered normal activity 4 day after cell separation, but the main secretory function was speculated to be reduced in culture.

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Expression of Vitellogenin Gene by Androgens in Rasinbow Trout, Oncorhynchus mykiss (웅성호르몬에 의한 무지개송어의 vitellogenin 유전자 발현)

  • 권혁추;윤종만;이종영
    • Journal of Aquaculture
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    • v.13 no.1
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    • pp.79-85
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    • 2000
  • The effects of estrogen and androgens on Vg gene expression were examined in primary hepatocyte culture and livers of the immature male trout. Specific primers of Vg cDNA were designed with already reported Vg gene nucleotide sequences. PCR product was sequenced and verified with Vg cDNA of rainbow trout. Total RNA was extracted from the cultured hepatocytes and livers of steroid-treated rainbow trout and then it was analyzed by reverse transcriptase- polymerase chain reaction (RT-PCR) analysis. The Vg mRNA and Vg protein synthesis were increased in rainbow trout in vivo and in vitro with E$_2$ and methyltestosterone (MT) There were dose and time-related effects of E$_2$ and MT on vitellogesis. Androgens such as progesterone androsterone and testosterone also stimulated Vg mRNA expression in vitro. The results show that androgens as well as E$_2$ can induce expression of Vg mRNA in trout in vivo and in vitro.

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Detection of Malignant Primary Hepatic Neoplasms with Gadobenate Dimeglumine (Gd-BOPTA) Enhanced T1-Weighted Hepatocyte Phase MR Imaging: Results of Off-site Blinded Review in a Phase-II Multicenter Trial

  • Constantino S. Pena;Sanjay Saini;Richard L. Baron;Bernd A. Hamm;Giovanni Morana;Roberto Caudana;Andrea Giovagnoni;Andrea Villa;Alessandro Carriero;Didier Mathieu;Michael W. Bourne;Miles A. Kirchin;Gianpaolo Pirovano;Alberto Spinazzi
    • Korean Journal of Radiology
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    • v.2 no.4
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    • pp.210-215
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    • 2001
  • Objective: To investigate the efficacy of gadobenate dimeglumine (GdBOPTA) enhanced MR imaging for the detection of liver lesions in patients with primary malignant hepatic neoplasms. Materials and Methods: Thirty-one patients with histologically proven primary malignancy of the liver were evaluated before and after administration of GdBOPTA at dose 0.05 or 0.10 mmol/kg. T1-weighted spin echo (T1W-SE) and gradient echo (T1W-GRE) images were evaluated for lesion number, location, size and confidence by three off-site independent reviewers and the findings were compared to reference standard imaging (intraoperative ultrasound, computed tomography during arterial portography or lipiodol computed tomography). Results were analyzed for significance using a two-sided McNemar's test. Results: More lesions were identified on Gd-BOPTA enhanced images than on unenhanced images and there was no significant difference in lesion detection between either concentration. The largest benefit was in detection of lesions under 1 cm in size (7 to 21, 9 to 15, 16 to 18 for reviewers A, B, C respectively). In 68% of the patients with more than one lesion, Gd-BOPTA increased the number of lesions detected. Conclusion: Liver MR imaging after Gd-BOPTA increases the detection of liver lesions in patients with primary malignant hepatic neoplasm.

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Subcellular Localization of Capsaicin-Hydrolyzing Enzyme in Rat Hepatocytes (Capsaicin 가수분해효소의 흰쥐 간세포내 소재확인)

  • Park, Young-Ho;Lee, Sang-Sup
    • YAKHAK HOEJI
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    • v.38 no.1
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    • pp.12-19
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    • 1994
  • Capsaicin(8-methyl-N-vanillyl-6-nonenamide) is the principal pungent component of Capsicum fruits. This work is directed to the capsaicin-hydrolyzing enzyme playing a key role in the rate limiting and critical step of capsaicin metabolism. In order to get precise information on the enzyme's subcellular location, rat liver homogenate was divided into six subcellular fractions by differential centrifugation technique: crude nuclear pellet, PNS(post nuclear supernatant) fraction, lysosomal pellet, cytosol, Tris wash fraction, micrisomes. Capsaicin-hydrolysing enzyme activity was analysed by high performance liquid chromatography(HPLC). This enzyme was found at the highest specific activity in the microsomal fraction and co-distributed with marker enzymes of the endoplasmic reticulum, NADPH-cytochrome c reductase and nucleoside diphosphatase. This is compatible with the result of ninhydrin color reaction of vanillylamine, primary metabolite of capsaicin hydrolysis, on thin layer chromatography(TLC). This enzyme is most active at pH $8.0{\sim}9.0$. Definite subcellular location of this enzyme will make it easy to proceed with further study.

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Molecular Basis of Drug Resistance: Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitors and Anaplastic Lymphoma Kinase Inhibitors

  • Yang, Sei-Hoon
    • Tuberculosis and Respiratory Diseases
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    • v.75 no.5
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    • pp.188-198
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    • 2013
  • Over the past decade, several kinase inhibitors have been approved based on their clinical benefit in cancer patients. Unfortunately, in many cases, patients develop resistance to these agents via secondary mutations and alternative mechanisms. To date, several major mechanisms of acquired resistance, such as secondary mutation of the epidermal growth factor receptor (EGFR) gene, amplification of the MET gene and overexpression of hepatocyte growth factor, have been reported. This review describes the recent findings on the mechanisms of primary and acquired resistance to EGFR tyrosine kinase inhibitors and acquired resistance to anaplastic lymphoma kinase inhibitors, primarily focusing on non-small cell lung carcinoma.