• 생약학회지
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• 제21권1호
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• pp.92-99
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• 1990

Effects of the methanol extract of Bupleuri Radix on primary cultured chicken embryonic brain cells, dorsal root ganglia (DRG) and rat hepatocytes were studied. The methanol extract of Bupleuri Radix at the concentration ranging from $10{\;}{\mu}g/ml\;to\;100{\;}{\mu}g/ml$ significantly recovered the cytotoxicity of rat hepatocytes induced by the treatment of galactosamine; at the concentration of $100\;{\mu}g/ml$, values of GOT and GPT in the culture medium were reduced by the 60% and 75%, respectively of those in the absence of the methanol extract of Bupleuri Radix. The addition of the methanol extract of Bupleuri Radix. into chicken embryonic brain cells which were cultured with a deficient medium significantly increased the number of cells promoting the neurite outgrowth. However, the methanol extract of Bupleuri Radix showed no effect on the activities of PDHC and acetylcholinesterase in primary cultured brain cells.

• Archives of Pharmacal Research
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• 제26권1호
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• pp.64-69
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• 2003

To analyze vasopressin-induced $Ca^{2+}$ increase in liver cells, rat hepatocytes were isolated and attached to collagen-coated cover slips. Using fura-2, a $Ca^{2+}$-sensing dye, changes in intracellular $Ca^{2+}$ concentration by vasopressin were monitored. Results in this communication suggested that vasopressin-induced $Ca^{2+}$ increase were composed of both $Ca^{2+}$ release from internal $Ca^{2+}$ stores and influx from the plasma membrane. The $Ca^{2+}$ influx consisted of two distinguishable components. One was dependent on the presence of vasopressin and the other was not. SK＆F96365 blocked vasopressin-induced $Ca^{2+}$ influx in a dose-dependent manner. Vasopressin-induced $Ca^{2+}$ release from internal stores diminished in a primary culture of hepatocytes according to the culture time. However, changes in vasopressin-induced $Ca^{2+}$ influx across the plasma membrane differed from changes in the $Ca^{2+}$ release from internal stores, suggesting two separate signalings from receptor activation to internal stores and to the plasma membrane.

• 대한약리학회지
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• 제32권3호
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• pp.399-406
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• 1996

Thioacetamide는 non-genotoxic 발암제로서 세포단백의 변형을 초래하는 것으로 알려져 있으며 이것을 단기간 처리하면 핵소체의 비대를 초래하게 된다. 본 연구에서는 thioacetamide를 처리한 간세포를 in vivo와 in vitro 상태에서 관찰하였다. In vivo상태로서 thioacetamide를 쥐의 복강에 7일간 주사하면 (50mg/kg), 핵소체 비대와 B23 및 MAP kinase와 같은 신호전달분자들이 증가하는 것을 관찰할 수 있었다. In vitro 상태로서 쥐의 간장을 collagenase로 분리하여 유전자 치료에 사용될 수 있는 배양조건으로 간세포를 배양하여 핵소체를 관찰한 결과 핵소체 비대가 현저하였으며, B23의 양도 증가하였다. 본 실험의 결과로 미루어 볼 때, 간세포는 핵소체 비대 수용능력이 약 100배 이상이라고 할 수 있으며, thioacetamide 처리에 의하여 라이보좀 생성과 핵소체 증가 능력이 증폭되어 나타나는 것으로 사료된다.

• 생약학회지
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• 제23권4호
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• pp.268-275
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• 1992

We studied to screen medicinal plants having hepatoprotective activity with the primary cultured rat hepatocytes intoxicated with carbon tetrachloride cytotoxicity. The lowest concentration and treatment time of carbon tetrachloride giving the greatest intoxication to the primary cultured hepatocytes were observed in 10mM and 60 minutes, respectively. GTP and GOT activity of culture broth of the primary cultured rat hepatocytes intoxicated by $CCl_4$ cytotoxicity at this condition were increased 135.9% and 178.3% compared with that of the primaries cultured hepatocytes not treated with $CCl_4$, respectively. This increased GPT activity was inhibited by glycyrrizin, which was known to have hepatoprotective activity, and the inhibition activity was dependent on the concentration of glycyrrhizin. Forty species among the extracts obtained from 117 species of medicinal plants were shown to have the hepatoprotective activity. Among these 40 species, Prunus persica, Scutellaria baicalensis, Astragalus membranaceus, Tribulus terrestris, Caragana chamlagu, Acanthopanax sessiliflorum and Achyranthes japonica were indicated a lower GPT activity than that of Glycyrrhiza uralensis containing glycyrrhizin and GPT activity of these were indicated 75.5%, 70.0%, 59.0%, 77.5%, 60.0%, 75.0% and 79.0%, respectively.

• Fisheries and Aquatic Sciences
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• 제1권1호
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• pp.24-29
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• 1998

Effect of extracellular calcium in vitellogenin (VTG) production in response to estradiol-17 $\beta$ $(E_2,\;2\times10^{-6}M)$ was examined in primary hepatocyte culture of rainbow trout, Onchorhynchus mykiss. Total calcium in estrogenized sera significantly increased, compared with the control, while diffusible calcium was insignificant. However, diffusible calcium in the incubation medium with $E_2$ was significantly reduced, compared with the control. The uptake of extracellular calcium by cultured hepatocytes signifIcantly increased 90 min after $E_2$ addition. Moreover, the accumulation of intracellular calcium increased in the cultures with $E_2$, regardless of the calcium concentrations in the incubation media. In addition, $E_2-primed $ VTG production was significantly decreased by withdrawal of E_2$ from the incubation medium. Moreover, VTG production by $E_2-primed$ hepatocytes was reduced by removing calcium from the incubation medium with or without $E_2$. These results suggest that the entry of extracellular calcium into the cytoplasm is an important step for VTG production in primary hepatocyte cultures in rainbow trout.

• 한국양식학회지
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• 제11권4호
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• pp.557-564
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• 1998

무지개송어의 초대배양 간세포에 있어서 Vitellogenin (VTG)의 합성 유도는 전기영동적 수법에 의해 행하였다. 간세포는 7일 동안 phositively charged dish를 이용한 배양으로 단층 확산되었다. 배양 7일의 간세포 생존율은 $E_2$의 유무에 따라 각각 20.7% 및 23.6%가 감소하였으며, DNA함유량도 각각 13.7% 및 14.0%가 감소하였다. $E_2$ 첨가군과 대조군간에는 생존율과 DNA 함유량에 있어서는 유의한 차이가 나타나지 않았다. 그리고, 총 단백질에 대한 VTG의 비율은 $E_2$$10^{-6}$ M의 첨가시에 최대치를 나타내었다. 그러나, 고농도($10^{-5}$M)에서는 오히려 감소하는 경향을 나타내었다.

• Toxicological Research
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• 제16권2호
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• pp.125-131
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• 2000

The ethanol extract of Rhus verniciflua Stokes (RVS), the Korean Lacquer tree, was subsequentely isolated and fractioned into two portions using distilled water (SED) and 99% ethanol (SEE) as elution buffers through silica gel column (4x28 em, 22 $\AA$. 28~200 mesh). To know the antioxidative effect of the RVS extracts, primary hepatocytes were exposed to hydroxyl radical generated by 20 mU/$m\ell$ glucose oxidase with SED or SEE for 4 hr. The addition of 100$\mu\textrm{g}$/$m\ell$ SED in culture medium showed good protection from glucose oxidase (GO)-mediated cytotoxicity of hepatocytes, showing approximately equivalent to control. When the hepatocytes were incubated with 100 $\mu\textrm{g}$/$m\ell$ SED or SEE only for 4 hr. the activities of cell-associated superoxide dismutase (SOD) and catalase were elevated up to 1.22 fold and 1.4 fold, respectively, compared to control. Further increase, 1.88fold in SOD activity or 1.64fold in catalase activity, was also observed when the hepatocytes were incubated with 100 units/$m\ell$ of commercial SOD or catalase for 4 hr. Moreover. the glucose oxidase-mediated cytotoxicity in cultured hepatocytes was generally reduced upon addition of lysate obtained from SED or SEE-stimulated hepatocytes in a dose-dependent manner. From these results, we suggest that, in cultured hepatocytes, RVS ethanol extract can efficiently reduce cytotoxicity induced by glucose oxidase and may increase the activity of cell-associated SOD and/or catalase, thereby preventing and/or scavenging superoxides and hydroxyl radicals in this experiment.

• KSBB Journal
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• 제7권4호
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• pp.278-283
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• 1992

쥐 간세포를 분리하고 여러가지 물질로 표면이 처리된 용기를 이용하여 배양하였다. 표면처리를 하지 않았을 경우와 collagen으로 처리하였을 경우 간세포는 monolayer형태를 이루며 자랐고, heparin과 hyaluronic acid의 경우에는 multilayer, 그리고 proteoglycan, dermatan sulfate(D. S.), BSA의 경우에는 hemispheroid, 그리고 표면처리를 하지 않은 Primaria는 spheroid형태를 형성하였다. 세포 생존도 면에서는 monolayer가 우수하였고, 간세포의 중요한 대사능이라고 볼 수 있는 암모니아의 제거능이나 albumin생산성 면에서는 hemispheroid 혹은 spheroid가 우수하였다.

• Journal of Microbiology and Biotechnology
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• 제11권3호
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• pp.423-427
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• 2001

Chitosan/alginate capsules were formed by electrostatic interactions and had appropriated mechanical strength, permeability to albumin, and stability to hepatocytes. Rat hepatocytes were isolated and immobilized in chitosan/alginate capsules. During the encapsulation process with hepatocyte, 10% of viability was decreased mainly due to the low pH of the chitosan solution. Among various capsule fabrication methods, the chitosan-alginate capsule showed the highest mechanical strength. Addition of collagen in the capsule with hepatocytes enhanced hepatic metabolism as well as the cell viability for 2 weeks of culture. The hepatocyte in the capsule without collagen decreased the viability to 10% for 2-week cultures.

• Journal of Microbiology and Biotechnology
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• 제15권1호
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• pp.7-13
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• 2005

Chitosan-alginate capsules were formed by electrostatic interactions and exhibited an appropriate mechanical strength, permeability, and stability for the culture of hepatocytes. Pig hepatocytes were isolated and hepatocyte spheroids formed and immobilized in chitosan-alginate capsules. An encapsulation procedure of 3 min and spheroid formation period of 24 h were the optimum conditions for the best liver functions. Pig hepatocytes with a cell density of $6.0{\tomes}10^6$ cells/ml in the capsules were found to be most suitable for application in a bioartificial liver support system. The encapsulated pig hepatocyte spheroids exhibited stable ammonia removal and urea secretion rates in a bioreactor for 2 weeks. Accordingly, chitosan-alginate encapsulated hepatocyte spheroids in a packed-bed bioreactor would appear to have potential as a bioartificial liver.