• Title/Summary/Keyword: primary hepatocytes

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Hepatoprotective Effects of Paecilomyces tenuipes Against Carbon Tetrachloride-induced Toxicity in Primary Cultures of Adult Rat Hepatocytes

  • Hyun, Sun-Hee;Jeon, Tae-Won;Lee, Sang-Kyu;Kim, Chun-Hwa;Seo, Young-Min;Kim, Ju-Hyun;Jeong, He-Min;Kang, Mi-Jeong;Lee, Jae-Sung;Jeong, Tae-Cheon
    • Toxicological Research
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    • v.23 no.4
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    • pp.301-309
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    • 2007
  • Paecilomyces tenuipes (PT), one of the Ascomycetes family, has been used for medicinal purposes due to its broad pharmacological activities. The present study was undertaken to investigate the hepatoprotective effects of PT water extracts against $CCl_4$-induced hepatotoxicity in primary cultures of adult rat hepatocytes. When the extract of PT was directly added into the culture medium at 1, 2, and 5 mg/ml, the extracts not only reduce the $CCl_4$-induced elevation of aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase, and lipid peroxide, but also protect cultured hepatocytes from $CCl_4$-induced reduction of reduced glutathione, glutathione reductase, glutathione-S-transferase, glutathione peroxidase, catalase and superoxide dismutase. In addition, the effects of PT water extracts on cytochrome P450 enzymes were relatively marginal, indicating that the hepatoprotective effects of PT extract against $CCl_4$-induced toxicity might not be due to the inhibition of $CCl_4$ activation. In conclusion, the PT extracts were effective in protecting against $CCl_4$ induced hepatotoxicity in hepatocyte cultures, at least in part, by scavenging free radicals, and by modulating enzyme systems involved in cellular oxidative stress.

Expression Patterns of Heat Shock Proteins in Primary Cultured Hepatocytes from Flounder (Paralichthys olivaceus)

  • Kim Woo Jin;Park Doo Won;Park Jung Youn;Kang Ho Sung;Kim Han Do
    • Fisheries and Aquatic Sciences
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    • v.2 no.1
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    • pp.85-92
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    • 1999
  • We examined the expression patterns of heat shock proteins in primary cultured hepatocytes from flounder (Paralichthys olivaceus) exposed to heat shock. The expression of hsp90, hsp70, hsp40, hsp30, and hsp27 was induced and major polypeptides were hsp70, hsp30 and hsp27. Northern blot analysis showed that expression of hsp70 was inhibited by transcriptional inhibitor actinomycin D, suggesting that expression of hsp70 gene is regulated at the transcriptional level. Prolonged exposure of cells to an elevated incubation temperature $(30^{\circ}C)$ induced the transient synthesis of hsp90, hsp70, hsp40, and hsp30 whereas maintenance of cells at a slightly higher incubation temperature $(32^{\circ}C)$ induced the continuous syntheses of these hsps. When cells were incubated at a higher temperatures $(35^{\circ}C\;or\;37^{\circ}C)$, the synthesis of hsps was almost similar to that of hsps in cells exposed to 32't except for the induction of hsp27 synthesis. These results that temperature and incubation time for optimum expression of each hsp during prolonged heat shock are different.

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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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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.

Antihepatotoxic and Antioxidant Activities of Polysaccharide Obtained from Cultured Mycelia of Ganoderma lucidum

  • Lee, June-Woo
    • The Korean Journal of Food And Nutrition
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    • v.32 no.5
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    • pp.417-424
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    • 2019
  • The purpose of this study was to observe the effects of the polysaccharide (GLP) obtained from the liquid cultured Ganoderma lucidum on the lipidperoxidation in a rat liver microsome and hepatotoxicity in the primary cultured rat hepatocytes. It is well known that the polysaccharide of Ganoderma lucidum exhibits hepatoprotective activity, antitumor activity etc., which many suggest a relationship to lipidperoxidation. The effect of GLP on $CCl_4-$ and galactosamineintoxicated cytotoxicity in the primary cultured rat hepatocytes were reduced the GPT value. In order to the estimate the effects of anti-lipidperoxidation of the polysaccharide, enzymatic and nonenzymatic reaction assays were performed, in vitro, in the rat liver microsome. An enzymatic lipidperoxidation reaction by $ADP+FeCl_3+NADPH$ and $CCl_4+NADPH$, GLP (1 mg/mL) inhibited 77.4% and 39.4%, respectively, and the nonenzymatic reaction displayed a 97.4% strongly inhibition. In the enzymatic and nonenzymatic inducers treated with GLP, the formation of malondialdehyde (MDA) progressively decreased by raising the GLP concentration. These results suggest that the anti-lipidperoxidation and radical scavenging activity of GLP may play an important part in the liver protection action.

Aluminium and Cadmium Interfere with the Estrogen Receptor Level in the Primary Culture of Hepatocytes in the Rainbow Trout Oncorhynchus mykiss

  • Hwang Un-Gi
    • Fisheries and Aquatic Sciences
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    • v.4 no.4
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    • pp.180-185
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    • 2001
  • Al and Cd-induced inhibition of vitellogenin (VTG) production was examined at the estrogen receptor (ER) level in rainbow trout Oncorhynchus mykiss hepatocytes. The binding of $[^3H]$ $estradiol-17\beta\;(E_2)$ to hepatocytes reached a plateau 3 days after addition of $E_2\;(2\times\;10^{-6} M)$to the medium. The binding activity was linearly reduced with the increased concentrations $(-10^{-5}\;M)$ of 4-hydroxy-tamoxifen (4-OHT) and specific binding linearly increased with the increased doses of $[^3H]\;E_2$, indicating that the radioligand bound to ER. Al $(-10^{-4}\;M)$and Cd $(10^{-6}\;M)$ as well as 4-OHT $(10^{-6}\;M)$ significantly reduced the $[^3H]\;E_2$-binding activity by $30­40\%$, while they completely inhibited VTG production. Al and Cd had no effect on $E_2-human$ $ER\alpha$ binding activity at any concentrations used $(-10^5\;nM\;each)$. These results suggested that Al and Cd inhibited VTG production in part by interfereing with the ER level. Inhibitory effects of these metals on the $E_z-dependent$ upregulation of ER activity are also discussed.

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Effects of copper sulfate poisoning on the fine structure of rat liver (유산동(硫酸銅) 중독(中毒)이 랫트 간장(肝臟)의 미세구조(微細構造)에 미치는 영향(影響))

  • Yoon, Hwa-joong;Lee, Sang-mog;Jang, Byoung-joon
    • Korean Journal of Veterinary Research
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    • v.30 no.1
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    • pp.85-91
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    • 1990
  • This study was designed in order to investigate the effect of copper sulfate to the ultrastructural changes of the hepatocytes in Sprague Dawley rats. The animals were administered with copper sulfate (10mg/kg B.W.), which was dissolved in normal saline. The solution was injected into abdominal cavity every day. The animals were sacrificed at the 6th, 12th, and 24th day from the beginning of administration. The specimens obtained from the liver were observed with electron microscope and significant changes were as follows. 1. A prominent dilatation and disruption of the cisternae of rough endoplasmic reticulum were recognized. Also, the detachment of membrane bound ribosomes was shown. 2. The proliferation of smooth endoplasmic reticulum and the depletion of glycogen particles were noted. 3. The increase of primary lysosomes and autophagic vacuoles was obserbed. 4. The dilatation of mitochondrial cristae was obserbed. And it was irregulary scattered in the stroma of mitochondria. 5. The atrophy of microvilli in the bile canaliculi and space of Disse was prominent. 6. Membrane of hepatocytes was damaged and significant hydrophic degeneration was obserbed in the perisinusoidal regions. 7. The damage of Fat-storing cells was more significant than that of hepatocytes.

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Effect of Growth Hormone on Vitellogenin Production by Estradiol-17$\beta$ in the Culture of Hepatocytes in the Rainbow Trout Oncorhynchus mykiss

  • Yeo In-Kyu;Mugiya Yasuo
    • Fisheries and Aquatic Sciences
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    • v.1 no.1
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    • pp.19-23
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    • 1998
  • Effects of pituitary and thyroid hormones on estradiol-induced vitellogenin (VTG) induction were electrophoretically examined in primary hepatocyte cultures of rainbow trout. Hepatocytes were precultured for 2 days and then estradiol-17 $\beta$ $(E_2,\;2 \times 10^{-6}M)$>, triiodothyronine $(T_3,\;10^{-8}-10^{-6}M)$, bovine growth hormone (bGH, 10-100 ng/ml), ovine prolactin (oPRL, 100-500 ng/ml), and pituitary extract (PE) of rainbow trout (0.75PE/dish) were added to the incubation medium. The hepatocytes were cultured for 7 more days. The addition of oPRL to the incubation medium was not effective in increasing VTG production at any concentrations. The addition of PE to the incubation medium with $E_2$ was not effective in increasing VTG production. The addition of bGH to the incubation medium with $E_2$ was not effective in increasing the rate of VTG production at concentrations of 10-50 ng/ml. However, a higher concentration of bGH, 100 ng/ml, increased VTG production. The various concentrations of $T_3$ were ineffective in stimulating VTG production. These results suggest that GH could be one of stimulus factors for VTG production in rainbow trout.

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