• KSBB Journal
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• v.11 no.4
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• pp.453-461
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• 1996

We compared the effects of two different systems of collagen matrix protein application on the survival and the biological functions of cultured primary hepatocytes. The rat liver primary hepatocytes were grown for approximately 40 days in vitro either on single collagen gel or between collagen sandwich gels. The morphological changes were observed for this culture period. While the hepatocytes grown on single gel began to die around at 7 days of culture, the cells grown between collagen gels still maintained their viability and began to die after 15 days. As markers for liver hepatic functions, we determined the biochemical activities of hepatocytes such as the secretions of albumin, fibronectin, fibrinogen, urea, and the reduction of secreted ammonia. We found that the rat hepatocytes cultured between collagen gels maintained fairly good biochemical functions than the hepatocytes cultured on single gel did. Therefore, the application of an extracellular matrix protein, collagen, in sandwich form was confirmed as a better choice for maintaining the functional hepatocytes culture for long term in vitro.

• Archives of Pharmacal Research
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• v.21 no.6
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• pp.718-722
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• 1998

Scopoletin (7-hydroxy-6-methoxycoumarin), a coumarin, was isolated from the aerial part of Solanum lyratum Thunb. by the activity-guided fractionation employing carbon te trachloride-intoxicated primary cultured rat hepatocytes as a screening system. Its hepatoprotective activity was first evaluated by measuring the release of glutamic pyruvic transaminase and sorbitol dehydrogenase from carbon tetrachloride-intoxicated rat hepatocytes into the culture medium. Scopoletin significantly reduced the releases of glutamic pyruvic transaminase and sorbitol dehydrogenase from the carbon tetrachloride-intoxicated primary cultured rat hepatocytes by 53% and 58%, respectively, from the toxicity in a dose-dependent manner over concentration ranges of 1mcM to 50mcM. Further studies revealed that at the concentration of 10mcM, scopoletin significantly preserved glutathione content by 50% and the activity of superoxide dismutase by 36% and also inhibited the production of malondialdehyde to the degree as seen in the control.

• Proceedings of the Ginseng society Conference
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• 1998.06a
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• pp.263-269
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• 1998

It is an established fact that most of the carcinogens implicate bay-region diol epoxides as the ultimate carcinogenic metabolites. These electrophiles react with nucleophilic sites in the cells to form abducts. It is the formation of carcinogenic-DNA adducts that is thought to initiate carcinogenesis. In our previous study we have reported chemopreventive property of Ginseng on 7,12-dimethylbenz (a)anthracene (DMBA) induced skin papillomagenesis in male Swiss albino mice. In this study we have examined the effect on formation of DMBA-DNA adducts in rat hepatocytes pretreated with ginseng. Primary cultures of rat hepatocytes were used. The cells wets treated with ginseng for 24 hrs and then with DMBA (iOn) for 18 hrs. Cells were then harvested, their DNA was isolated and analyzed by P)2 labelling. A significant reduction in the levels of DMBA-DNA adduces (adducts/108 nucleotides) was observed in all cultures pretreated with ginseng. The viability of cells was not affected by pre-treatment with ginseng. Our finding suggests that ginseng block or suppresses the events associated with chemical carcinogenesis by inhibiting metabolic activation of the carcinogens.

• YAKHAK HOEJI
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• v.50 no.4
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• pp.287-292
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• 2006

Protective effects of the water extract of Protaetia brevitarsis larva against $CCl_4-induced$ toxicity were investigated in primary cultures of adult rat hepatocytes. The extract used in these studies contained several minerals, fatty acids and amino acids. Treatment of hepatocyte cultures with the extract provided a significant protection from the increased LDH activity induced by $CCl_4$. The results demonstrated that the extract may have the protective effect against $CCl_4-induced$ toxicity in hepatocyte cultures.

• Toxicological Research
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• v.3 no.2
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• pp.73-80
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• 1987

Primary cultures of adult rat hepatocytes were used to study the cytotoxicity of D-galactosamine. Hepatocytes were isolated by a collagenase perfusion technique and maintained as monolayers in serum-free medium on collagen-coated culture dishes. Treatment of galactosamine to the culture markedly inhibited the uptake of ${\alpha}$-aminoisobutyric acid (AIB) inducible with glucagon and dexamethasone. At0.1 mM of galactosamine, AIB uptake was inhibited significantly when treated for 12 hr. At higher doses (0.25, 0.5 and 1.0mM), a significant inhibition was noticed after 1 hr exposure. Generally the magnitude of the inhibition was related to the dose and treatment time of galactosamine. Treatment of galactosamine also produced a dose- and treatment time-related suppression of the tyrosine aminotransferase (TAT) induction caused by dexamethasone. Meanwhile, uptake of ouabain was not affected by the treatment of galactosamine. The viability of the hepatocytes was decreased only slightly by the treatment of galactosamine; more than 87% of the cells excluded tryphane blue when treated 1 mM galactosamine for 12 hr. Galactosamine induced depressions of AIB uptake and TAT activity were prevented by the simultaneous addition of uridine to the culture. D-Galactosamine, cytotoxicity, hepatocytes culture, ${\alpha}$-aminoisobutyric acid uptake, tyrosine aminotransferase.

• KSBB Journal
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• v.7 no.4
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• pp.278-283
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• 1992

Rat hepatocytes were isolated and cultured on the petri dishes treated with various coating materials. Untreated or collagen coated petri dish gave monolayer culture of hepatocyte and proteoglycan, dermatan sulfate, and BSA treated petri dish gave hemispheroid. The untreated Primaria petri dish gave spheroid type of hepatocyte, and heparin and hyaluronic acid treatment gave multilayers. To sustain high cell viability, monolayer cultured hepatocytes was more useful, while it was found that the hemispheroid or spheroid type hepatocytes was more active in the hepatic functions such as ammonia metabolism and albumin synthesis.

• Journal of the Korean Society of Food Science and Nutrition
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• v.20 no.1
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• pp.40-45
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• 1991

The release of hepatic triglyceride lipase from cultured rat hepatocytes and its hormonal regulation were studied. The activity of lipase released into the medium in the presence of heparin was increasing during 24 hours on the 2nd of culture while this was 10% in the absence of heparin as compared with the lipase activity in the presense of heparin. When hepatocytes were cultured with anti-hepatic triglyceride lipase lgG the lipase activity was supp-ressed by 92% The results suggest that the enzyme relaeased into culture medium is identical to hepatic triglyceride lipase which can be released only in the presence of heparin the model of release being similar to that of lipoprotein lipase from adipocytes. The addition of monensin to the medium resulted in The inhibition of lipase secretion by 61% Insulin enhanced lipase activity only 20% whereas dexamethasone suppressed the activity by 44% These data inidica-ted that hepatic triglyceride lipase is secreted and released from hepatocytes in the presence of heparin and its secretion is regulated by hormones.

• BMB Reports
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• v.28 no.6
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• pp.533-537
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• 1995

The detection with lucigenin under physiological conditions is selective for ${\cdot} O_{2}^{-}$, for it can be accepted that lucigenin indicates actual intramembranal $\cdot O_{2}^{-}- formation$. Lucigenin chemiluminescence (CL) was elicited from the plasma membrane (PM) only by addition of reduced pyridine nucleotide. NADPH was preferred to NADH in PM and hepatocytes. This specificity was masked by $NAD(P)^+$ inhibition. The half maximum rate of CL increase was obtained with 1.5 ${\mu}m$ NADH or 55 ${\mu}m$ NADPH in hepatocytes and 6 ${\mu}m$ NADH or 30 ${\mu}m$ NADPH in plasma membranes. Measurement of these NADPH values required the presence of a NADPH-regenerating system. With NADPH the maximal rate obtained was 10 fold higher than with NADH. NADPH and NADH could produce CL when having access from either side of the membrane. They seemed to react with the identical acceptor because NADH-induced CL was also inhibited by $NADP^+$. The characteristics of ${\cdot}O_{2}^{-}-formation$ produced by pyridine nucleotide will be discussed.

• Toxicological Research
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• v.5 no.1
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• pp.9-15
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• 1989

The effect of butylated hydroxytoluene (BHT) and its major metabolite, 3, 5-di-tert-butyl-4-hydroxybenzoic acid (BHT-acid) on the uptake of taurocholate into hepatocytes was studied using the primary culture of rat hepatocytes. Hepatocyte were isolated by an in situ collagenase perfusion technique and maintained as a monolayer in serum-free meadia for 24 hours before use. The uptake of taurocholate was saturable with an apparent Km of 12.8+2.8 MuM and Vmax of 0.18+0.01 nmol/mg/min. Both BHT and BHT-acid inhibited the hepatocellular uptake of taurocholate when they were added to the culture.

• YAKHAK HOEJI
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• v.37 no.5
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• pp.499-503
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• 1993

Betaine, a major component of Lycii Fructus, was evaluated for its anti-hepatotoxic activity on carbon tetrachloride-induced hepatotoxicity in primary cultured rat hepatocytes. Betaine was found to attenuate carbon tetrachloride-induced hepatotoxicity both morphologically and biochemically. Typical hepatocyte necrosis due to carbon tetrachloride seemed to be reduced by 50 to 500 $\mu{M}$ of betaine under microscopical observation. The value of glutamic pyruvic transaminase released from the hepatocytes into the medium significantly decreased as betaine concentration increased. Betaine also significantly elevated the reduced activities of some enzymes, cytochrome P-450, 7-ethoxycoumarin-0-deethylase and glutathione-S-transferase, involved in xenobiotic metabolism due to carbon tetrachloride-induced hepatotoxicity. These results demonstrate a possible hepato-protective role of betaine against fatty liver that could be easily induced by carbon tetrachloride.