• 한국임상수의학회지
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• 제32권4호
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• pp.289-294
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• 2015

Fucoidan은 갈조류로부터 추출되는 황산다당류로 다양한 생리학적 활성을 갖고 있다. 본 연구의 목적은 LPS로 자극한 돼지 말초혈액 단핵구세포(PBMCs)의 NO 생산에 있어 fucoidan의 효과를 검토하고, 이러한 효과가 iNOS의 발현과 AP-1의 활성화와 관련이 있는지를 조사하는데 있다. LPS 무처치 돼지 PBMCs에서 fucoidand의 처리는 NO 생산과 AP-1활성에 대해 효과를 보이지 않았다. 또한 iNOS와 AP-1의 mRNA 발현도 fucoidan 처치에 의해 영향을 받지 않았다. 그러나 LPS로 자극한 PBMCs에서는 NO 생산과 AP-1의 활성 그리고 iNOS와 AP-1의 mRNA 발현이 현저하게 증가하였다. 이와 같은 LPS에 의한 돼지 PBMCs의 NO 생산과 AP-1 활성증가는 fucoidan 첨가에 의해 감소되었다. 또한 fucoidan은 LPS에 의한 iNOS와 AP-1의 mRNA 발현증가도 억제시켰다. 이상의 결과는 fucoidan이 LPS 자극 돼지 PBMCs에서 iNOS 발현과 AP-1 활성의 억제와 함께 NO 생산을 하향 조절함으로써 항염증효과를 나타내는 것으로 사료되었다.

• Journal of Ginseng Research
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• 제39권3호
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• pp.189-198
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• 2015

Background: Antiallergic effect of 20(S)-protopanaxatriol (PPT), an intestinal metabolite of ginseng saponins, was investigated in guinea pig lung mast cells and mouse bone marrow-derived mast cells activated by a specific antigen/antibody reaction. Methods: Increasing concentrations of PPT were pretreated 5 min prior to antigen stimulation, and various inflammatory mediator releases and their relevant cellular signaling events were measured in those cells. Results: PPT dose-dependently reduced the release of histamine and leukotrienes in both types of mast cells. Especially, in activated bone marrow-derived mast cells, PPT inhibited the expression of Syk protein, cytokine mRNA, cyclooxygenase-1/2, and phospholipase $A_2$ ($PLA_2$), as well as the activities of various protein kinase C isoforms, mitogen-activated protein kinases, $PLA_2$, and transcription factors (nuclear factor-${\kappa}B$ and activator protein-1). Conclusion: PPT reduces the release of inflammatory mediators via inhibiting multiple cellular signaling pathways comprising the $Ca^{2+}$ influx, protein kinase C, and $PLA_2$, which are propagated by Syk activation upon allergic stimulation of mast cells.

• BMB Reports
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• 제28권5호
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• pp.387-391
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• 1995

In order to investigate whether phospholipase C (PLC) activity in oat celIs is regulated by Gprotein, we have characterized PLC in plasma membranes of oat tissues. To identify the purified plasma membrane, $K^+$-stimulated, $Mg^{2+}$-dependent ATPase activity was measured. The activity of ATPase was shown to be proportional to the concentration of membrane protein. To examine the PLC activity regulated by G-protein, we used the inside-out and outside-out plasma membrane mixture isolated from the oat cells. The plasma membrane mixture showed higher PLC activity than the one of the outside-out plasma membrane. This suggests that PLC activity is located at the cytoplasmic surface of plasma membrane. PLC activity in plasma membrane mixture was dependent on $Ca^{2+}$ with maximum activity at 100 ${\mu}m$ $Ca^{2+}$ and it was inhibited by 1 mM EGTA. Using Sep-pak $Accell^{TM}$ Plus QMA chromatography, we found that inositol 1,4,5-trisphosphate ($IP_3$) was produced in the presence of 10 ${\mu}m$ $Ca^{2+}$. The PLC activity in the membrane was enhanced by an activator of G-protein ($GTP{\gamma}S$) and not by an inhibitor ($GDP{\beta}S$). This indicates that a G-protein is involved in the activation of PLC in the plasma membrane of oat cells.

• Journal of Photoscience
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• 제9권2호
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• pp.86-89
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• 2002

Blue light activates proton pump, and creates electrical gradient across the plasma membrane and drives $K^{+}$ uptake in stomatal guard cells. In this presentation, we provide evidence for regulatory mechanisms of the pump and the identification of blue light receptor. The pump is shown to be the plasma membrane H$^{+}$- ATPase and is activated through phosphorylation of the C-terminus. Phosphorylation occurred and 14-3-3 protein bound to the phosphorylation site. The binding of 14-3-3 protein was required for the H$^{+}$-ATPase activation. We also found that phot1 phot2 double mutant does not respond to blue light but other mutants respond to blue light by stomatal opening. However, all these mutants are capable of stomatal opening in the presence of fusicoccin, an activator of the H$^{+}$-ATPase. These results suggest that both photl and phot2 act as blue light receptors in guard cells.d cells.

• 한국약용작물학회지
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• 제18권1호
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• pp.1-8
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• 2010

Inonotus obliquus is one of the immune-regulatory substances and is recognized to play the role in the metabolic process of inflammation, allergy and immuntiy. The purpose of this study was to evaluate the effects of water extracts of Inonotus obliquus (IOW) on the liver lymphocyte immune function in the Sprague-Dawley male rats treated with carbon tetrachloride ($CCl_4$) to induce liver damage. Rats were fed with each experimental diet and water for 4 weeks. We found that effects of IOW on interferon-gamma (IFN-$\gamma$), signal transducer and activator of transcription 1 (STAT1), phospho-signal transducer and activator of transcription 1 (pSTAT1) and GATA-binding protein 3 (GATA-3) were decrease in vivo. Interleukin-4 (IL-4), STAT6, pSTAT6 and T-box expressed in T-cells (T-bet) decreased significantly lower in $CCl_4$+IOW group than the $CCl_4$ group. Our data indicated that cytokine protein production were increased in $CCl_4$ group and $CCl_4$+IOW group. As a result of this study, we assume that IOW fed could regulate the immuno-modulating functions through regulate the cytokine production capacity activated by liver damage.

• Journal of Ginseng Research
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• 제48권2호
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• pp.211-219
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• 2024

Background: Recently, plant-derived exosome-like nanoparticles (PDENs) have been isolated, and active research was focusing on understanding their properties and functions. In this study, the characteristics and molecular properties of ginseng root-derived exosome-like nanoparticles (GrDENs) were examined in terms of skin protection. Methods: HPLC-MS protocols were used to analyze the ginsenoside contents in GrDENs. To investigate the beneficial effect of GrDENs on skin, HaCaT cells were pre-treated with GrDENs (0-2 × 10⁹ particles/mL), and followed by UVB irradiation or H₂O₂ exposure. In addition, the antioxidant activity of GrDENs was measured using a fluorescence microscope or flow cytometry. Finally, molecular mechanisms were examined with immunoblotting analysis. Results: GrDENs contained detectable levels of ginsenosides (Re, Rg1, Rb1, Rf, Rg2 (S), Gyp17, Rd, C-Mc1, C-O, and F2). In UVB-irradiated HaCaT cells, GrDENs protected cells from death and reduced ROS production. GrDENs downregulated the mRNA expression of proapoptotic genes, including BAX, caspase-1, -3, -6, -7, and -8 and the ratio of cleaved caspase-8, -9, and -3 in a dose-dependent manner. In addition, GrDENs reduced the mRNA levels of aging-related genes (MMP2 and 3), proinflammatory genes (COX-2 and IL-6), and cellular senescence biomarker p21, possibly by suppressing activator protein-1 signaling. Conclusions: This study demonstrates the protective effects of GrDENs against skin damage caused by UV and oxidative stress, providing new insights into beneficial uses of ginseng. In particular, our results suggest GrDENs as a potential active ingredient in cosmeceuticals to promote skin health.

• Radiation Oncology Journal
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• 제24권1호
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• pp.58-66
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• 2006

목적: 방사선에 의한 신장손상은 궁극적으로 신장 섬유화로 인한 신부전으로 나타나며 여기에는 세포외기질의 변화가 동반된다. 방사선 신장손상에서 신사구체 상피세포의 역할을 알아보기 위하여 방사선에 의한 세포외기질과 연관된 여러 유전자 발현의 변화를 알아보고자 하였다. 대상 및 방법 : 백서 사구체 상피세포 (rat glomerular epithelial cell: GEpC) 에 6 MV 선형가속기 (Siemens, USA)를 이용하여 0, 2, 5, 10, 20 Gy 의 단일 방사선량을 조사한 후 각각 6, 24, 48, 72 시간에 시료를 채취하였다. Northern blot, Western blot, Zymography를 이용하여 fibronectin (Fn), plasminogen activator inhibitor-1 (Pai-1), matrix metalloproteinases-2, 9 (MMP-2, 9), tissue inhibitor of matrix metallproteinase-2 (TIMP-2), tissue-type plasminogen activator (t-PA), Urokinase-type plasminogen activator (u-PA)의 발현을 측정하였다. 결과: GEpC 에 대한 10 Gy 단일 방사선 조사후 24 시간부터 Fn mRNA 가 유의한 증가를 나타냈으며 48 시간에 측정한 Fn 단백질은 5, 10 Gy 의 방사선량에서 유의하게 증가되었다. 방사선조사에 의해서 Pai-1 유전자의 발현도 mRNA 및 단백질 단계에서 증가되었으며, 특히 10Gy 조사 후 24, 48 시간에 측정한 mRNA 의 증가는 통계적으로 유의하였다. GEpC에 방사선조사 후 24 시간에 측정한 MMP-2 활성형은 방사선량에 따라 증가하였으나 통계학적 유의성은 없었다. 그밖의 MMP-9, TIMP-2, t-PA 와 u-PA 는 아무런 변화를 나타내지 않았다. 결론 : 방사선에 의하여 GEpC에서 세포외 기질과 관련된 유전자 발현의 번화가 관찰되었으며 이는 방사선 신장 손상에 GEpC가 관여함을 나타낸다.

• Journal of Microbiology and Biotechnology
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• 제16권5호
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• pp.695-703
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• 2006

The dissolved oxygen level of any cell culture environment has a critical effect on cellular metabolism. Specifically, hypoxia condition decreases cell viability and recombinant protein productivity. In this work, to develop CHO cells producing recombinant protein with high productivity, mammalian expression vectors containing a human tissue-type plasminogen activator (t-PA) gene with hypoxia response element (HRE) were constructed and stably transfected into CHO cells. CHO/2HRE-t-PA cells produced 2-folds higher recombinant t-PA production than CHO/t-PA cells in a $Ba^{2+}-alginate$ immobilized culture, and 16.8-folds in a repeated batch culture. In a non-aerated batch culture of suspension-adapted cells, t-PA productivity of CHO/2HRE/t-PA cells was 4.2-folds higher than that of CHO/t-PA cells. Our results indicate that HRE is a useful tool for the enhancement of protein productivity in mammalian cell cultures.

• BMB Reports
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• 제37권3호
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• pp.320-334
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• 2004

StMBF1 (Solanum tuberosum multiprotein bridging factor 1) is a plant member of the MBF1 family of transcriptional co-activators. In an attempt to understand the role of StMBF1, we analyzed its interaction with plant transcription factors of the homeodomain-leucine zipper (Hd-Zip) family, a group of proteins with a typical leucine zipper motif adjacent to a homeodomain. StMBF1 is able to interact in vitro with the Hd-Zip protein Hahb-4 both in the presence and absence of DNA. Upon binding, StMBF1 increases the DNA binding affinity of Hahb-4, and of another plant homeodomain containing protein from the GL2/Hd-Zip IV family, HAHR-1. The biological role of interactions is discussed in this paper.

• BMB Reports
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• 제52권4호
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• pp.265-270
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• 2019

We investigated whether SIRT1 is associated with reactive oxygen species (ROS) accumulation during CK2 downregulation-mediated senescence. SIRT1 overexpression suppressed ROS accumulation, reduced transcription of FoxO3a target genes, and nuclear export and acetylation of FoxO3a, which were induced by CK2 downregulation in HCT116 and MCF-7 cells. Conversely, overexpression of a dominant-negative mutant SIRT1 (H363Y) counteracted decreased ROS levels, increased transcriptional activity of FoxO3a, and increased nuclear import and decreased acetylation of FoxO3a, which were induced by CK2 upregulation. CK2 downregulation destabilized SIRT1 protein via an ubiquitin-proteasome pathway in human cells, whereas CK2 overexpression reduced ubiquitination of SIRT1. Finally, the SIRT1 activator resveratrol attenuated the accumulation of ROS and lipofuscin as well as lifespan shortening, and reduced expression of the DAF-16 target gene sod-3, which were induced by CK2 downregulation in nematodes. Altogether, this study demonstrates that inactivation of the SIRT1-FoxO3a axis, at least in part, is involved in ROS generation during CK2 downregulation-mediated cellular senescence and nematode aging.