• KSBB Journal
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• 제7권4호
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• pp.271-277
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• 1992

쥐 간세포를collagenase perfusion method에 의해 분리한 뒤 collagen coated dish와floating collagen membrane에서 일차배양하여 간세포의 분화기능을 조사하였다. 두 경우 모두 간세포의 생존율은 5일 이후부터 점차 감소하였거나 간세포에 의한 암모니아 처리기능과 albumin생성기능은 약 7일간 유지되었다. 또 이러한 분화기능에 유지는 ollagen coated dish보다floating collagen membrane이 유리한 것으로 나타났다.

• Toxicological Research
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• 제15권1호
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• pp.89-93
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• 1999

2,3,7,8-Tetrachlorodibenzo-p-dioxin(TCDD) is known to induce cytochrome p450 1A1 and to activate c-Src kinase and p21 Ras. This study examined the molecular interactions of adaptor proteins including Shc, Grb2, and Sos in rat primary hepatocytes and their relationship to the induction of CYP1A1 by TCDD. TCDD induced CYP1A1 level and EROD activity in a dose-dependent mode. Sos/Grb2 association isincreased by TCDDㅑㅜ a dose dependent mode. Tyrosine phosphorylated Shc, mainly p152, onloads to Grb2/Sos complex upon TCDD stimulation. The electrophoretic mobility shift of Sos is showed by TCDD. These results indicate that TCDD modulated the molecular interaction features of adaptor compoes proteins including Shc, Grb2, and Cnl in early signaling pathway of TCDD-mediated CYP 1A1 induction of rat primary hepatocyte.

• 한국식품영양과학회지
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• 제21권5호
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• pp.496-501
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• 1992

간세포내 두가지 microsome효소인 HMG-CoA reductase와 cholesterol-$7{\alpha}$-hydroxylase는 콜레스테롤 합성과 담즙산으로의 분해의 조절효소임은 이미 알려진 사실이다. 본 실험에서는 콜레스테롤의 대사산물인 4종류 담즙산들이 간세포내의 콜레스테롤 합성 및 HMG-CoA reductase활성에 미치는 효과를 조사하였다. 간세포의 배양에서 간세포로의 담즙산 흡수는 배지내 농도(1mM~10mM)와 배양시간(1/2~3hr)의 차이에 따라 비례적으로 증가하였다. 콜레스테롤 합성저해에 대한 담즙산의 효과는 배지내 담즙산의 농도 및 배양시간에 따라 크게 감소하였다. 분리시킨 microsome내 HMG-CoA reductase의 활성에 대한 담즙산의 저해효과는 insulin 투여에 의하여 효소활성을 촉진시킨 경우에서도 뚜렷하였으며 콜레스테롤 합성 역시 저하되었다. 분리시킨 간세포막 내 $Na^+$,$K^+$-ATPase의 활성은 1.8~2.5mM정도의 배지내 담즙산 농도까지 증가함을 보였으나 cholic acid 흡수는 상기 효소의 활성에 거의 영향을 주지 않는 것으로 나타났다. 이러한 이유는 아직 불분명하나 1차 대사산물인 cholic acid의 흡수는 단순확산에 의한 것으로 사료되며 이에 대한 더 많은 연구가 요구된다.

• Fisheries and Aquatic Sciences
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• 제1권2호
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• pp.153-158
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• 1998

Effects of cyclic (c) AMP and G-protein related reagents (3-isobutyl-l-methyxanthine (IBMX), Forskolin (FSK), cholera toxin (CTX), and pertussis toxin (PTX≫ on estradiol-17$\beta$ induced vitellogenin (VTG) induction were examined in primary hepatocyte cultures in rainbow trout Oncorhynchus mykiss. The addition of IBMX, FSK, or CTX to the incubation medium markedly increased VTG production, while PTX was not effective in stimulating the production. It is well known that cAMP regulates phosphorylation and dephosphorylation through mediation of protein kinase A. These results suggest that VTG production is highly dependent on cAMP state in hepatocytes because of its highly phosphorylated nature.

• 약학회지
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• 제34권5호
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• pp.341-347
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• 1990

The anti-hepatotoxic activity of Panax ginseng was studied using galactosamine (GalN)-induced cytotoxicity in primary cultured rat hepatocytes. Panax ginseng was fractionated into dammarane glycosides and protein fractions. The dammarane glycosides was further fractionated into panaxadiol and panaxatriol glycosides fractions. The protein fraction was further fractionated into four groups according to the molecular weight; larger than 10,000 dalton, between 5,000 and 10,000 dalton, between 1,000 and 5,000 dalton and between 500 and 1,000 dalton. A significant lowering action on the elevated glutamicpyruvic transaminase (GPT) activity in the culture medium of hepatocytes treated with 1.5 mM GalN was noticed with all four protein fractions studied at the concentration of both $50\;{\mu}g/ml$ and $100\;{\mu}g/ml$. However, the effect of dammarane glycosides fractions was not significant. It was noted that the addition of $100\;{\mu}g/ml$ of protein fractions smaller than 5,000 dalton significantly enhanced the syntheses of protein and RNA in the damaged hepatocytes induced by the treatment of 1.5 mM GalN. Dammarane glycosides fractions significantly enhanced protein synthesis at the concentration of $100\;{\mu}g/ml$ in the damaged hepatocytes by treatment of 1.5 mM GalN.

• 한국양식학회지
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• 제11권3호
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• pp.311-317
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• 1998

Estradiol-17${\beta}$($E_2$)에 의한 Vitellogenin (VTG) 함성에 미치는 Cu 및 Zn의 영향을 무지개송어의 배양 간세포를 이용하여 조사하였다. 간세포는 2일간 배양 한 후, E하(2) ($2{\times}10^6$ M)와 동시에 Cu ($2{\times}10^{-5}$~$10^{-4}$M) 또는 Zn ($10^{-5}$~$10^{-3}$M)을 배양액에 첨가하여 5일간 배양하였다. VTG 합성률은 총 단백질에 대한 VTG의 배율로 나타내었다. CU 및 Zn의 첨가는 배양 간세포의 생존율에는 영향을 미치지 않았다. 그러나, Cu의 첨가에 사용된 모든 농도에서, Zn의 첨가에는 농도 의존적으로 감소하여(10^{-3}$M에서 유의하게 감소하였다. 이러한 감소는, Zn의 제거시에는 회복되었으나, Cu에서는 회복되지 않았다. 그리고, Cu (10^{-4}$M의 첨가 시에 Ca (1.8 mM) 농도를 2.5 및 5.0 mM로 증가시켜도 Cu의 작용을 저해하지 못하였다. 이러한 결과로 보아, Cu 및 Zn은 간세포에서 합성되는 다른 단백질 보다 VTG 합성에 더 깊이 관여하는 것으로 추정된다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XIII)
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• pp.316-316
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• 2003

• Applied Microscopy
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• 제13권1호
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• pp.13-30
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• 1983

[ $1-{\beta}-D-Arabinofuranosylcytosine$ ](ara-C), which is a pyrimidine nucleoside analog is cytotonic to mammalian cells in culture and is active in vitro and in vivo against a variety of DNA viruses. The precise mechanism of action of ara-C has not been determined, although ara-C is thought to act as an antimetabolite, interfering with the synthesis of deoxyribonucleic acid(DNA). Cytosine arabinoside originally seemed to act principally by inhibiting the conversion of cytidine to deoxytidine, thus inhibiting DNA synthesis. But recent data suggest that effects upon DNA polymerase and effects via incorporation into DNA and RNA may well be of equal importance. The author have demonstrated the effect of cytosine arabinoside on the hepatocytes of albino mice treated with ara-C, observing changes in the cytoplasmic organelles of the hepatocytes. A total of 120 healthy male albino mice were divided into the control and ara-C treated groups. The animals of the ara-C group were given 10mg. per kg of body weight of mouse ara-C in physiological saline solution and the animals of control group were given physiological saline solution, intraperitoneally. After an administration of ara-C or physiological saline solution, the animal were killed at. interval of 6, 12, and 24 hours. The specimens, which were obtained from the left anterier lobe of the liver, were stained with uranyl acetate and lead citrate and observed with JEM 100B electron microscope. The results were obtained as follow: A pronounced dilatation, sacculation and fragmentation of the cisterane of rough endoplasmic reticulum with dissociation of membrane bound-ribosomes, disaggregation of free ribosomes in the cytoplasm, proliferation of the smooth endoplasmic reticulum associated with depletion of glycogen paracles, atrophies of Golgi complex, production of numerous lipid droplets, and formation of antophagic vacuoles, multivesicular bodies and residual bodies are recognized in the hepatocytes of ara-C treated mice. Consequently it is suggested that cytosine arabinoside would induce a changes of the cytoplasmic organelles of the hepatocytes in albino mice.

• 대한한의학회지
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• 제19권1호
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• pp.145-164
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• 1998

The purpose of this study is to evaluate the effect of Injinchunggantang-derivative on proliferation of hepatocyte in rats. Cell viability is studied by MTI assay. The gene related to cell replication such as p53, waf1, bcl-2 and $bcl-_{X_L}$ is quantitized by quantitative RT-PCR and the proteins coded by these genes are studied by Western blotting. The results are as follows. 1. The hepatocytes cultured in medium with lnjinchunggantang-derivative showed better viability compared with control grroup in MTI assay, and the hepatocytes cultured in medium with the Injinchunggantang-derivative-and-ethanol-mixed group showed better viability than the hepatocytes cultrued in 10% ethanol culture medium(control group), noting that Injinchunggantang-derivative has protective effect on hepatocyte injury. There was no dose- and time-dependence. 2. In quantitative RT-PCR, i) Bel-2 gene increased significantly both in Injinchunggantang-derivative group and in Injinchunggantang-derivative-and-ethanol-mixed group, while it showed no significant increase or decrease in other group. ii) $Bcl-_{X_L}$ gene increased significantly in Injinchunggantang-derivative group as well as in Injinchunggantang-deri vative-and-ethanol -mixed group. iii) P53 gene showed no significant increase or decrease in hepatocytes cultured in medium with 10% ethanol and in hepatocytes cultured in medium with Injinchunggantang-derivative-and-ethanol-mixed group, suggesting that 10% ethanol induced cell toxicity, thus increased p53 gene expression. iv) Wafl gene showed no significant increase or decrease in hepatocytes cutured in medium with Injinchtrnggantang-derivative, while increased in hepatocytes cultured in medium with 10% ethanol and in hepatocytes cultured in medium with Injinchtrnggantang-derivative-andethanol-mixed group, suggesting that 10% ethanol induced cell toxicity increased wafl gene expression. 3. In the study on protein by western blotting, the band of bcl-2 and $bcl-_{X_L}$ were widened in Injinchtrnggantang-derivative group. Especially the amount of $bcl-_{X_L}$ increased significantly compared with other groups. But in the study on p53 and wafl, there was no significant difference among those groups. Above study shows that Injinchunggantang-derivative has good effect on cell viability and that the genes resistant to cell death such as bcl-2 and $bcl-_{X_L}$ are induced by Injinchunggantang-derivative to resist to cell death by toxic agent And this is reconfirmed in protein study using' western blotting: These results suggest that Injinchunggantang-derivative has inhibitory effect on cell death as well as protective effect on hepatocyte. Therefore this prescription is recommended in various liver diseases such as chronic liver disease and-induced hepatic injury.

• Fisheries and Aquatic Sciences
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• 제5권4호
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• pp.251-257
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• 2002

Effects of bisphenol-A (BPA) on vitellogenin (VTG) synthesis and estrogen-estrogen receptor $(E_2- ER)$ binding activity were examined in primary hepatocyte cultures of rainbow trout, Oncorhynchus mykiss. Hepatocytes were precultured for 2 days and then $estradiol-17\beta\;(E_2,\;2\times10^{-6}M)\;BPA\;(10^{-5}-10^{-8}M)$ and/or 4-hydroxy-tamoxifen $(4-OHT,\;10^{-6} M)$ were simultaneously added to the incubation medium. Hepatocytes were cultured for 5 more days and then spent medium was analyzed by SDS-PAGE for VTG production. The addition of BPA to the incubation medium had no effect on the viability of hepatocytes in the culture. On the other hand, BPA increased VTG production in a concentration-dependent way and a significant increment occurred at BPA concentrations greater than $10^{-6}$M. Although VTG was increased by the addition of $E_2\;(2\times10^{-6}\;M)\;or\;BPA\;(10^{-5}M)$, its were reduced by a simultaneous 4-OHT $(10^{-6}\;M)$ addition. BPA inhibited $E_2-human$ ER binding activity by $72\%$ at $10^{-5}$ M of BPA. These results suggested that BPA induced VTG synthesis by BPA-ER binding activity in the hepatocyte of rainbow trout.