• Korean Journal of Veterinary Research
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• v.38 no.3
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• pp.518-524
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• 1998

신장 근위세뇨관세포들은 사구체에서 여과된 물질의 재흡수, 분비 및 대사에 관여하는 여러 호르몬들의 수용체들을 가지고 있다. 이들중에서 dopamine(DA)과 angiotensin II(ANG II)가 $Na^{+}/H^{+}$ 상호운반계 조절에 중요한 역할을 하고 있다. 본 연구는 초대배양한 토끼 신장 근위세뇨관세포의 $Na^+$ uptake에 있어서 DA과 ANG II의 상호관계를 알아보고자 실시하였다. DA은 농도의존적으로 $Na^+$ uptake를 유의성 있게 억제하였다($10^{-6}M$ ; $83.2{\pm}7.2%$, $10^{-3}M$ ; $67.2{\pm}3.8%$ vs. control)(p<0.05). $DA_1$ 작동제(SKF 38393, $10^{-6}M$)는 대조군의 $81.4{\pm}6.7%$ 까지 $Na^+$ uptake를 유의성 있게 억제하였으나(p < 0.05) $DA_2$ 작동제는 영향을 미치지 않았다. $DA_1$ 길항제(SCH 23390, $10^{-6}M$)에 의해 DA의 $Na^+$ uptake 억제효과는 차단되었으나 $DA_2$ 길항제(spiperone, $10^{-6}M$)에 의해서는 영향을 받지 않았다. DA과 대조적으로 $10^{-11}M$ ANG II는 $AT_1$ 수용체를 통하여 대조군의 $120.7{\pm}4.9%$까지 $Na^+$ uptake를 유의성 있게 촉진하였다. (p < 0.05). DA 및 $10^{-11}M$ ANG II를 병합처리하였을 때 DA은 농도의존적으로 ANG II에 유도된 $Na^+$ uptake 촉진효과를 유의성 있게 차단하였다(p<0.05). 한편 ANG II에 의해 유도된 $Na^+$ uptake촉진작용은 $DA_1$ 또는 $DA_2$ 작동제에 의해 차단되었으나 DA에 의한 차단 효과는 $DA_1$ 및 $DA_2$ 길항제를 병합처리하였을 때만 반전되었다. 결론적으로 DA은 $DA_1$ 수용체를 통하여 $Na^+$ uptake를 억제하였으나 ANG II에 의한 $Na^+$ uptake 촉진작용의 억제에는 $DA_1$ 및 $DA_2$ 수용체 모두가 관여하였다.

• Korean Journal of Veterinary Research
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• v.38 no.3
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• pp.525-534
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• 1998

신장 근위세뇨관세포들은 사구체에서 여과된 물질의 재흡수, 분비 및 대사에 관여하는 여러 호르몬들의 수용체들을 가지고 있다. 이들중에서 norepinephrine(NE)과 angiotensin II(ANG II)는 $Na^{+}/H^+$ 상호운반계를 조절함으로써 혈압조절에 관여하는 것으로 알려져 있으나 이들의 상호관계에 대해선 연구보고가 많지 않다. 본 연구는 초대배양한 토끼신장 근위세뇨관세포를 이용한 $Na^+$ uptake 실험을 통하여 NE이 어떠한 수용체를 통하여 $Na^{+}/H^+$ 상호운반계를 조절하는지 그리고 이러한 작용에 있어서 NE과 ANG II의 상호관계를 알아보고자 실시하였다. NE(>$10^{-9}M$)은 $Na^+$ uptake를 유의성 있게 증가시켰다($10^{-9}M$ NE : $27{\pm}4%$ increase vs. Control;p < 0.05). $\alpha$ 길항제(phentolamine, $10^{-10}M$)는 NE($10^{-9}M$)에 의해 유도된 $Na^+$ uptake를 유의성 있게 차단하였으나 (phentolamine+NE : $29{\pm}5%$ inhibition vs. NE ; p〈 0.05), ${\alpha}_1$ (pra-zosin, $10^{-10}M$) 및 ${\alpha}_2$ 길항제(yohimbine, $10^{-10}M$)는 부분적으로 차단하였다. ${\beta}$ 길항제(propra-nolol, $10^{-10}M$)도 역시 NE에 의해 유도된 $Na^+$ uptake를 유의성 있게 차단하였으나(propranolol+NE : $24{\pm}6%$ inhibition vs. NE ; p< 0.05), ${\beta}_1$(atenolol, $10^{-10}M$) 및 ${\beta}_2$ 길항제(butoxamine, $10^{-10}M$)는 부분적으로 차단하였다. 이러한 결과들은 NE에 의해 유도된 $Na^+$ uptake 증가작용은 ${\alpha}$(${\alpha}_1$ 및 ${\alpha}_2$ )와 ${\beta}$(${\beta}_1$ 및 ${\beta}_2$) 수용체 모두를 통하여 일어난다는 것을 시사해주고 있다. ANG II($10^{-11}M$) 또는 NE(${\alpha}_1$, ${\alpha}_2$, ${\beta}_1$, ${\beta}_2$ 작동제) 단독처리군의 $Na^+$ uptake는 대조군에 비해 유의성 있게 증가하였으나 (ANG II : $23{\pm}9%$ increase vs. Control; p < 0.05), 병합처리시 상승작용은 나타나지 않았다. ${\alpha}$ 또는 ${\beta}$ 길항제 처리시 NE 및 ANG II에 의해 유도되었던 $Na^+$ uptake 증가는 유의성 있게 차단되었다(phentolamine+NE+ANG II : $25{\pm}3%$ inhibition, propranolol+NE+ANG II : $24{\pm}6%$ inhibition vs. NE+ANG II, respectively ; p〈 0.05). 이 결과들은 $Na^+$ uptake에 있어서 ${\alpha}$(${\alpha}_1$ 및 ${\alpha}_2$)와 ${\beta}$(${\beta}_1$ 및 ${\beta}_2$) 수용체와 ANG II의 관련성을 시사해 준다. 결론적으로 토끼 신장 근위세뇨관세포에서 NE은 ${\alpha}_1$, ${\alpha}_2$, ${\beta}_1$ 및 ${\beta}_2$ 수용체를 통하여 $Na^+$+ uptake를 증가시켰으며 이들 수용체는 ANG II $Na^+$ uptake 증가작용에 관여하였다.

• Korean Journal of Plant Tissue Culture
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• v.25 no.1
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• pp.33-36
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• 1998

Guzmania was propagated through in vitro culture of lateral shoots. When new shoots grown in greenhouse were cut and cultured in vitro, contamination rate was very high at about 80% in the first stage of in vitro culture. Among cytokinin treatments for agar medium, 2.0 mg/L BA was most effective for shoot multiplication, and those with 0.5 mg/L kinetin and 0.5~1.0 mg/L BA were favorable for shoot multiplication. BA was more effective for shoot multiplication than kinetin, and shoot multiplication was more enhanced when 2.0 mg/L BA was combined with 0.1~0.5 mg/L IAA than 2.0 mg/L BA alone. The medium with 2.0 mg/L BA and 0.1 mg/L IAA showed the highest rate of shoot multiplication with about 8.7 in shoot number, and those with 2.0 mg/L BA and 0.5~1.0 mg/L IAA also resulted in high multiplication of shoots. Shoots were multiplicated more in liquid rotation culture(80 rpm) with the medium containing 0.5 mg/L BA and 0.1 mg/L IAA than liquid stagnating and solid cultures. Regenerated shoots formed roots very favorably in the medium supplemented with 2.0 mg/L IBA.

• FLOWER RESEARCH JOURNAL
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• v.18 no.2
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• pp.83-86
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• 2010

This study was conducted to develop in vitro propagation techniques of new cultivar, 'Greenstar', bred by Highland Agriculture Research Center. The multiple shoot induction and plant growth of in vitro plant were analyzed by MS media concentration (1/2 MS, 1 MS and 2 MS medium), plant growth regulators and its proper concentration; CPPU [Forchlorfenuron(N-(2-chloro-4-pridyl)-3-phenylurea) (0, 0.5, 1.0 and $2.0mg{\cdot}L^{-1}$), thidiazuron [(TDZ), (0.01, 0.1, 0.5 and $1.0mg{\cdot}L^{-1}$), zeatin (0, 0.5, 1.0 and $2.0mg{\cdot}L^{-1}$), and BA [6-benzylaminiopurine(BA), (0, 0.5, 1.0 and $2.0mg{\cdot}L^{-1}$)] in MS (3% sugar and 0.8% agar with pH 5.7) media. The highest number of induced shoots, leaves and roots were shown in 1/2 MS medium concentration. On the 1/2 MS medium, shoot numbers, shoot length, leaf numbers and root numbers were 11.0, 1.9 cm, 24.7, and 8.0, respectively. On the absence of CPPU in the 1/2 MS medium, shoot length and root numbers was greater than CPPU treatment, but the highest number of shoots was induced by the $2.0mg{\cdot}L^{-1}$ of CPPU concentration in 1/2MS medium. TDZ, zeatin, and BA treatments were not effective on the induction of multiple shoot in vitro culture. As a result, in vitro culture of new Saxifraga fortunei, 'Greenstar' with $2.0mg{\cdot}L^{-1}$ of CPPU in 1/2 MS medium was most effective for the rapid multiplication.

• Journal of Life Science
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• v.28 no.6
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• pp.718-725
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• 2018

Toll-like receptor 4 (TLR4) has been implicated in cell proliferation and apoptosis in several types of cancer. In this study, the impact of TLR4 activation on apoptotic cell death in gynecologic cancers induced by lipopolysaccharide (LPS) was investigated. Cervical cancer cell lines were produced from isolated surgical specimens supplied by Paik Hospital. The primary cultures of normal myometrium and gynecologic cancers, including cervical, endometrial, and ovarian cancers, were used to examine the differences in morphological characteristics between normal and cancerous cells. A reverse transcription polymerase chain reaction analysis was used to determine the relative expression levels of TLR4 gene involved in apoptosis-associated signaling in cervical cancer cells. The cancer cell colonies showed a tendency to reach high levels of confluency compared with normal cells. In addition, an enhanced growth rate and loss of contact inhibition were observed in gynecologic cancer cells compared with normal cells (doubling times of 16.6 hr vs. 26 hr, respectively). The expression level of ITGA5, an alpha-5 integrin marker, was upregulated in normal myometrial cells, but this tendency was not exhibited in cervical cancer cells. Furthermore, p53 tumor suppressor gene expression was upregulated, whereas TLR4 and caspase-3 gene expressions were downregulated in cervical cancer cells. Notably, the expression levels of TLR4 and caspase-3 were increased significantly in LPS-treated cancer cells compared with those in non-LPS-treated cells. These results suggest that the TLR4-mediated caspase-dependent apoptotic signaling pathway could be suggested as a therapeutic target for the treatment of gynecologic cancers, including cervical cancers.

• Korean Journal of Pharmacognosy
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• v.23 no.4
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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.

• Journal of the Korea Institute of Military Science and Technology
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• v.16 no.3
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• pp.390-397
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• 2013

Primary-cultured immune cells are widely used in research to elucidate the mechanism of inflammation including chemotaxis, production of reactive oxygen species, cytokine release and antigen presenting. Mice are one of the species of experimental animals commonly used for such studies. Immune cells can be isolated and cultured from various organs such as bone marrow, peritoneal cavity, lung, spleen. For elaborated experimental studies, immune cells should be elicited with inflammatory substances or proliferated in vitro with special media. This paper details methods of obtaining immune cells from various organs of mice and investigating immune mechanism using isolated immune cells. It contains standard protocols of isolating and culturing immune cells from bone marrow, peritoneal cavity and lymphoid organs. It also covers the methods of investigating immune mechanism such as ELISA, western blotting, confocal microscopy and ELISPOT assay. With the works in this study, we established the standardized isolation and analysis methods of primary-cultured immune cells.

• Korean Journal of Ichthyology
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• v.12 no.3
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• pp.173-179
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• 2000

Effects of Estradiol-$17\beta(E_2)$ and 2, 4-dichlorophenoxy acetic acid (2, 4-D) on vitellogenin(VTG) production were investigated in primary hepatocyte culture of olive flounder, Paralichthys olivaceus. Highest survival rate of hepatocyte were observed at $27^{\circ}C$, which markedly declined equal to 50% of those of $15^{\circ}C$. Vitellogenin production peaked at the concentration of $10^{-6}M\;E_2$. No effect was observed on VTG production at various concentrations of 2, 4-D. However, a low concentration of 2, 4-D (ie, $10^{-8}M$) only appeared increased VTG production. $E_2$ or $10^{-8}M$ 2, 4-D-primed VTG production was markedly inhibited by the addition of $10^{-6}M$ tamoxifen to the culture medium(P<0.01). Inhibition was not affected by combinational treatment with $10^{-6}M$ $E_2$ and $10^{-6}M$ 2, 4-D. These results from the current investigation suggest that 2, 4-D mimics $E_2$, but the mechanism of reaction in inducing the $E_2$ receptor are different in VTG production in oliver flounder hepatocytes.

• Journal of Bio-Environment Control
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• v.24 no.3
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• pp.167-172
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• 2015

The aim of this study was to develop an effective production system of blueberry plants by using tissue culture technique. Murashige and skoog medium (MS) and woody plant medium (WPM) were compared for shoot formation of highbush blueberries. Also medium supplemented with zeatin/2-isopentenyl adenine (2iP)/benzyl aminopurine (BA) (1, 2/10, 15/4, $6mg{\cdot}L^{-1}$)and zeatin/2iP/BA (0.5/10, 15/$0.05mg{\cdot}L^{-1}$) as plant growth regulators to determine the effect of shoot formation and shoot proliferation, respectively. The shoot explants cultured on WPM showed higher shoot formation rates, more number of nodes, and longer root length than those on MS medium during the primary culture. Shoots were not formed when the explants were cultured on the medium without plant growth regulators or on only BA. The shoot explants cultured on the medium supplemented with 2iP showed low rates of shoot formation. On the other hand, zeatin was the most effective for shoot formation and growth of the explants. Also influence of different cytokinins (zeatin, 2iP) on the shoot proliferation of subcultured shoot explants was studied. There was no significant difference among the different concentrations of zeatin in the rate of shoot formation and number of shoots. However at higher concentration of zeatin, number of nodes was increased, and shoot length was shorted. The proper concentrations of zeatin for shoot propagation in subculture were found to be $0.5mg{\cdot}L^{-1}$ and $1mg{\cdot}L^{-1}$.

• Environmental Analysis Health and Toxicology
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• v.9 no.1_2
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• pp.19-23
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• 1994

The tetrazolium dye, 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT), is reduced by live but not dead cell, and this reaction is used as the end point in a rapid drug screening assay. It can also be used for accurate determinations of drug sensitivity but only if a quantative relationship is established between cell number and MTT-formazan production. Several conditions were examined to devise an in vitro assay method in primary cultured hepatocytes, such as optimum wavelength, optimal MTT concentration, optimal incubation time, and cell density.