• Toxicological Research
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• 제8권2호
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• pp.285-302
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• 1992

Human and rat hepatocytes were isolated by nonperfusion method and cultured for longer than 5 days. Human liver biopsy sample and rat liver were used as hepatocyte source. Several physical and chemical factors which were influencing on hepatocyte isolation procedure were examined and a batch isolation procedure was established for small size sample of rat liver. Isolated hepatocytes showed normal morphlologica characteristics in microscopy and electron microscopical examinations and a morphologica response to phalloidin. Isolated cells were cultured as a monolayer and proven to have intact morphological characteristics for longer than 15 days. Because human liver sample is harder and tighter compared with rat liver, a standard procedure for rat hepatocytes was slightly modified to reduce mechanical damage. Similarly with rat hepatocytes, isolated human hepatocytes showed a normal morphological characteristics and could be cultured for longer than 15days. Human and rat hepatocytes were examined on their functional integrities including cytochrome-P450 related enzyme activity and it's inducibility, hormonal inducibility of AIB uptake and TAT activity, albumin synthesis, DNA synthesis, cellular protein maintenance. In all parameters used in the present study, human and rat hepatocytes showed normal functional characteristics.

• 한국식품위생안전성학회:학술대회논문집
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• 한국식품위생안전성학회 2002년도 춘계학술발표대회 및 심포지움
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• pp.147-147
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• 2002

This experimental study was carried out to investigate on the safety of Nog-yong aqua-acupuncture solution in primary culture of adult rat hepatocytes. Primary culture of adult rat hepatocytes has been considered as a ideal model for toxicological studies because cultured hepatocytes maintained many liver-specific functions. In this research, we investigated the effects of Nog-yong aqua-aqupunture(1-10$\mu\textrm{g}$/ml) on cytotoxicites in primary culture of adult rat hepatocytes using LDH release assays. Hepatic glutathione level. glutathione-S-transferase activity, and albumin synthesis were not affected by treatment with Nog-yong aqua-aqupuntur alone. Nog-yong aqua-aqupuncoure solution(0.5-$10{\mu}\textrm{g}$/ml) on cytotoxicites in primary culture of adult rat hepatocytes using LDH release not significantly affected normal functional charaterists.

• 생약학회지
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• 제26권1호
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• pp.51-56
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• 1995

To devise an in vitro screening method for antihepatotoxic activity, $CCl_4-induced$ cytotoxicities in primary cultures rat hepatocytes were examined. When rat hepatocytes were intoxicated with 0.5, 1.0 or 1.5 mM $CCl_4$ for 1.5, 3 or 19hr, in order of LDH>GOT>GPT release form hepatocytes was increased in a dose-dependent manner. Treatment with 1.5 mM $CCl_4$ for 1.5 hr showed maximum increase in activity of LDH, GOT or GPT released in the medium compared with the control. At this experimental condition, well known antihepatotoxic substances, glycyrrhizin and silybin markedly inhibited $CCl_4-induced$ cytotoxicities. These results demonstrated that the screening method using $CCl_4-induced$ injury in primary cultured rat hepatocytes might be suitable in vitro assay for antihepatotoxic activity.

• 약학회지
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• 제42권1호
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• pp.108-113
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• 1998

G009, a polysaccharide isolated from the mycelia of Ganoderma lucidum IYO09, showed a hepatoprotective activity against $CCl_4$ induced cytotoxicity in primary cu ltured rat hepatocytes. Incubation of $CCl_4$-intoxicated hepatocytes with G009 significantly reduced the levels of glutamic pyruvic transaminase and sorbitol dehydrogenase released from hepatocytes in the medium. G009 showed antioxidative effect by elevating the activities of glutathione reductase and superoxide dismutase, and the content of glutathione in $CCl_4$-intoxidcated primary cultured rat hepatocytes. Furthermore, G009 significantly elevated glutathione-S-transferase activity in $CCl_4$-intoxicated primary cultured rat hepatocytes. G009 also reduced the production of malondialdehyde, a byproduct of lipid peroxidation. From these results, it could be concluded that G009 exerted hepatoprotective activity against $CCl_4$-induced cytotoxicity through antioxidation.

• 생약학회지
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• 제25권4호통권99호
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• pp.388-394
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• 1994

80% Methanol extracts of 44 traditional drugs used for the treatment of liver diseases or tonic effects were screened for anti-hepatotoxic activity by in vitro assay using $CCl_4-induced$ cytotoxicity in primary cultured rat hepatocytes. $CCl_4-induced$ cytotoxicity was evaluated by determination of LDH, GOT or GPT activity in the medium. Rehmaniae Radix Preparata and Gelantina nigra inhibited the release of LDH, GOT or GPT from $CCl_4-treated$ hepatocytes. Gibotii Rhizoma and Eucommiae Cortex showed inhibitory effect on release of LDH from normal hepatocytes as well as $CCl_4-treated$ hepatocytes. Eucommiae Cortex and Lili Bulbus decreased release of GOT and LDH from normal hepatocytes, respectively. Astragali Radix inhibited release of GPT in $CCl_4-treated$ hepatocytes. Phlomidis Radix, Imperatae Rhizoma, Cistanchis Herba, Broussonetiae Fructus, Asparagi Tuber, Trigonellae Semen and Polgonati Rhizoma inhibited release of LDH from $CCl_4-treated$ hepatocytes. Among 44 traditional drugs, most of them released LDH, GOT or GPT at the dose of 1 mg/ml in normal hepatocytes, and Drynariae Rhizoma, Acanthopanacis Cortex, Longanae Arillus, Atratylodis Rhizoma and Ecliptae Herba increased $CCl_4-induced$ cytotoxicity.

• Archives of Pharmacal Research
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• 제18권4호
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• pp.289-292
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• 1995

Icariside II, a flavonol glycoside, was isolated from the aerial part of Epimedium Koreanum Nakai by the anti-hepatotoxic acitivity guided fractionation technique employing $CCl_4-in-toxicated$ primary cultured rat hepatocytes as an assay system. Its anti-hepatotoxic activity was evaluated by measuring activity of glutamic pyruvic transaminase released from the $CCl_4-in-toxicated$ primary cultured rat hapatocytes. Icariside II significantly reduced the activity of glutamic pyruvic transaminase released from the $CCl_4-in-toxicated$ primary cultured rat hepatocytes and resulted in 78% recovery of the toxicity at the concentration of $200{\;}\mu\textrm{m}$. The anti-hepatotoxic activity of icariside II on the $CCl_4-in-toxicated$ primary cultured rat hepatocytes was as potent as that of silybin.

• Toxicological Research
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• 제4권1호
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• pp.37-45
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• 1988

Primary cultures of adult rat hepatocytes were used to study in vitro cytotoxic effects of T-2 toxin on liver cells. When T-2 toxin was added to the culture, a significant depression of the hormonal induction of ${\alpha}$-aminoisobutyric acid (AIB) uptake and tyrosine aminotransferase (TAT) activity was observed. However, T-2 toxin did not affect the uptake of ouabain into hepatocytes. Protein synthesis was inhibited by T-2 toxin, but RNA synthesis was not severely affected. The inhibitory effects of T-2 toxin on protein synthesis was diminished rapidly with culture time and the hepatocytes culture maintained control level of protein synthesis within 24 hrs.

• Biomolecules & Therapeutics
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• 제4권3호
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• pp.271-274
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• 1996

The antihepatotoxic activity of Bezoar Bovis and Moschus was investigated by in vitro assay method using galactosamine and carbon tetrachloride-induced cytotoxicity in primary-cultured rat hepatocytes. The antihepatotoxic activity was evaluated by measuring the level of glutamate pyruvate transaminase and sorbitol dehydrogenase which were released from the necrotic hepatocytes to the culture medium. In galactosamine-intoxicated hepatocytes, the chloroform fraction of Bezoar Bovis reduced the level of glutamate pyruvate transaminase and sorbitol dehydrogenase resulting in 65% and 59% protection, respectively. The n-Hexane fraction of Moschus resulted in 45% and 40% protection, respectively in this system. In the case of carbon tetrachloride-intoxicated rat hepatocytes, Bezoar Bovis did not have significant effect and only the aqueous fraction of Moschus showed 42% and 40% protection, respectively.

• Toxicological Research
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• 제2권1호
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• pp.1-7
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• 1986

Procarcinogen induced DNA single-strand breaks and unschduled DNA synthesis were measured in primary rat hepatocytes culture. For DNA single-strand breaks assay, rat liver DNA was prelabeled by injection 3H-thymidine during the peak of DNA synthesis following partial hepatectomy. Hepatocytes were isolated from the rat 2 weeks after surgery by a collagenase perfusion techinique and maintained as monolayers in serum free medium on collagen-coated culture dishes. DNA sigle-strand breaks were measured by the alkaline elution techinique.

• Biomolecules & Therapeutics
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• 제12권3호
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• pp.185-188
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• 2004

The purpose of this study is to evaluate the effect of cigarette smoke condensate (CSC) on toxification/detoxification metabolic pathway in primary cultured rat hepatocytes. We measured the activities of cytochrome P450 monooxygenases (CYP450s) and UDP-glucuronyltransferase, sulfotransferase and glutathione-S-transferase in CSC-treated rat hepatocytes. CSC significantly increased the activities of hepatic CYP4501A1 and CYP4501A2 to 7.5 fold and 1.6 fold respectively, compared with control level. However, CSC did not affect the activities of conjugation enzymes. We a1so examined if treatment of CSC could change thc cytotoxicity of acetaminophen (AA) through modulation of metabolizing enzymes. In rat hepatocytes, pretreatment with CSC potentiated the cytotoxicity of AA. This result indicates that potentiation of AA toxicity by CSC pretreatment may be related to induction of CYP4501A1 and CYP4501A2.