• KSBB Journal
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• 제24권4호
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• pp.397-402
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• 2009

본 연구에서는 몇 가지 생물공학 기반 기술로서 이미 한방 화장품의 소재로 사용되고 있는 쇠비름 에탄올 추출액의 tyrosinase 저해, collagen 합성, 항염증 기대 효과, 항노화 대한 효능을 알아보기 위하여 B16F10 mouse melanoma cell, NIH3T3 mouse embryonic fibroblast, MCF-7 human breast adenocarcinoma cell에 쇠비름 추출액을 처리하여 실험하였다. 실험 결과, 쇠비름 에탄을 추출액 (0.5mg/mL)은 tyrosinase 발현을 억제하여 멜라닌 합성을 억제하며, 농도 의존적으로 NIH3T3 mouse fibroblast 세포의 collagen 합성을 촉진시켰다. 또한 2.0 mg/mL 농도의 쇠비름 추출액은 목단피보다 더욱 효과적으로 cytokine (TNF-$\alpha$)에 의한 NF-${\kappa}B$ 활성을 억제시켰다. Doxorubicin에 의한 과노화에서도 효과적인 항노화 작용을 하는 것으로 나타났다. Tyrosinase 합성을 억제하므로 미백에 대한 효과와 세포의 생장을 도와 collagen 합성을 촉진함으로서 노화방지에 효과적인 것으로 사료되며, TNF-$\alpha$에 의한 NF-${\kappa}B$의 활성화를 저해함으로서 염증반응 억제 효과도 기대 할 수 있다.

• 대한약학회:학술대회논문집
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• 대한약학회 2001년도 Proceedings of the Pharmaceutical Society of Korea
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• pp.169.2-170
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• 2001

• BMB Reports
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• 제49권2호
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• pp.99-104
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• 2016

The nuclear non-histone protein high mobility group box (HMGB) 1 is known to having an inhibitory effect on the repair of DNA damaged by the antitumor drug cisplatin in vitro. To investigate the role of HMGB1 in living cells, we studied the DNA repair of cisplatin damages in mouse fibroblast cell line, NIH-3T3. We evaluated the effect of the post-synthetic acetylation and C-terminal domain of the protein by overexpression of the parental and mutant GFP fused forms of HMGB1. The results revealed that HMGB1 had also an inhibitory effect on the repair of cisplatin damaged DNA in vivo. The silencing of HMGB1 in NIH-3T3 cells increased the cellular DNA repair potential. The increased levels of repair synthesis could be "rescued" and returned to less than normal levels if the knockdown cells were transfected with plasmids encoding HMGB1 and HMGB1 K2A. In this case, the truncated form of HMGB1 also exhibited a slight inhibitory effect.

• 한국해양바이오학회지
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• 제7권1호
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• pp.28-34
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• 2015

We examined cell regeneration efficiency of brown marine algae living in Jeju coast for search of a novel therapeutic device with cutaneous wound healing materials. The five algae were collected and compared with epidermal growth factor (EGF) as a positive control in the assays of cell proliferation and cell migration of NIH3T3 fibroblast cells. Among the 80% methanol extracts of these brown algae, the two algal extracts from Ishige foliacea and Colpomenia bullosa showed the proliferative effects of the cells similar to the effect of EGF. Besides it was found that Colpomenia bullosa extract significantly enhanced cell migration of NIH3T3 cell. In the study, therefore, we confirmed that the Colpomenia bullosa extract improved proliferation of NIH3T3 cell and a potential candidate for cultaneous wound healing.

• Parasites, Hosts and Diseases
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• 제48권1호
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• pp.23-33
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• 2010

An understanding of the nature of the immune response to asexual erythrocytic stages of malaria parasites will facilitate vaccine development by identifying which responses the vaccine should preferentially induce. The present study examined and compared the immune responses of NIH mice in either single or mixed infections with avirulent (DK) or virulent (DS) strains of Plasmodium chabaudi adami using the ELISA test for detecting and measurement of cytokines and antibody production. In both single and mixed infections, the study showed that both cell- and antibody-mediated responses were activated. In all experiments, an early relatively high level of IFN-$\gamma$ and IgG2a during the acute phase of the infection, and later elevation of IL-4 and IgG1, suggested that there was a sequential Th1/Th2 response. However, in the avirulent DK strain infection a stronger Th1 response was observed compared to the virulent DS strain-infection or in mixed infections. In the virulent DS infection, there was a stronger Th2 response compared to that in the DK and mixed infections. The faster proliferation rate of the virulent DS strain compared to the DK strain was also evident.

• BMB Reports
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• 제49권7호
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• pp.370-375
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• 2016

Ras oncoproteins are small molecular weight GTPases known for their involvement in oncogenesis, which operate in a complex signaling network with multiple effectors. Approximately 25% of human tumors possess mutations in a member of this family. The Raf1/MEK/Erk1/2 pathway is one of the most intensively studied signaling mechanisms. Different levels of regulation account for the inactivation of MAP kinases by MAPK phosphatases in a cell type- and stimuli-dependent manner. In the present study, using three inducible Ras-expressing NIH/3T3 cell lines, we demonstrated that MKP3 upregulation requires the activation of the Erk1/2 pathway, which correlates with the shutdown of this pathway. We also demonstrated, by applying pharmacological inhibitors and effector mutants of Ras, that induction of MKP3 at the protein level is positively regulated by the oncogenic Ras/Raf/MEK/Erk1/2 signaling pathway.

• BMB Reports
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• 제28권2호
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• pp.87-93
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• 1995

To investigate the biological functions of the EGF-like domain of mouse betacellulin (BTC), mouse BTC(33-80), a 48-residue peptide corresponding to the EGF-like domain, was synthesized by stepwise solidphase methods using a 9-fluorenylmethoxycarbonyl (Fmoc) strategy. The homogeneity of synthetic mouse BTC(33-80) was confirmed by analytical reversed phase (RP)-HPLC, amimo acid analysis, and fast atom bombardment mass spectrometer (FAB-MS). Three disulfide bond pairings of synthetic mouse BTC(33-80) were established by amino acid analysis of cysteine-containing fragments derived from thermolytic digestion. These were consistent with the pairings of EGF and transforming growth factor ($TGF-{\alpha}$). The EGF-Iike domain of mouse BTC showed equipotent activity in both EGF-receptor binding on A-431 epidermoid carcinoma cells, and mitogenesis on NIH-3T3 fibroblast cells, as compared with authentic h-EGF. Results suggest that the EGF-Iike domain of BTC plays a significant role in mitogenic activity with an EGF-receptor mediated system.

• Applied Biological Chemistry
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• 제47권2호
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• pp.202-207
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• 2004

어성초 분말을 포함하는 차가버섯 발효 조성물의 물 추출물이 인체 위암 세포 AGS 및 대장암 세포 HCT-15, 그리고 정상세포 NIH3T3 fibroblast의 세포 생육에 미치는 영향을 검토하였다. 세포독성 실험은 세포수 count와 MTT 방법으로 측정하였다. 어성초 분말을 포함하는 차가버섯 발효 조성물의 제조는 차가버섯과 어성초 분말을 혼합하고, 여기에 대두 발효 미생물원을 다시 혼합시켜 습도 $50{\sim}60%$,온도 $30{\sim}37^{\circ}C$에서 30일 정도 발효시킨 후 건조 시킨 분말에서 5% 물 추출물을 얻어 동결건조 시켜 실험에 제공하였다. MTT assay 방법에서 차가버섯 발효 조성물의 수용성 추출물 0.16, 0.4, 0.8, 1.6 및 4.0 mg/ml 첨가 농도에서 AGS 세포 생육은 13, 25, 40, 67 및78% 억제 되었으며, HCT-15세포 생육은 22, 40, 50, 69및 76%씩 각각 억제되었다. 그러나 동일한 실험조건에서 정상 세포수 NIH3T3은 86% 이상의 생존율을 나타내었다. 차가버섯 발효 조성물의 수용성 추출물은 인체 대장암 세포주 HCT-15와 위암 세포수 AGS에 대해 생육억제 작용이 강한 반면, 정상세포 NIH3T3에 대해서는 세포 독성을 거의 나타내지 않아 가장 바람직한 암예방 또는 항암식품 개발가능성을 제시하였다.

• Journal of Ginseng Research
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• 제15권3호
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• pp.183-187
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• 1991

Effect of ginseng hairy and native roots on body absorption, fecal and urinary excretion of Ingested radiostrontium were investigated in the mouse(NIH-strain, male) treated with or without pre-feeding of each ginseng soluble fraction. The test groups were fed with basic diet supplemented with 1% each ginseng soluble fraction for 7 darts before the radiostrontium were administered by intragastric intubation. In the groups of treated with soluble fraction from ginseng hairy roots, the radioactivities of fecal and urinary excretion increased about 15% over than that of control groups and the whole body retention were about 38%. In the groups of treated with soluble fraction from native ginseng roots, the radioactivities of fecal and urinary excretion increased about 25% over than that of control groups and the whole body retention were about 28%. Also, the levels of radiostrontium accumulation retained significantly the higher percent in skeletons than in other organs.

• Clinical and Experimental Reproductive Medicine
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• 제33권3호
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• pp.189-198
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• 2006

목적: 본 연구진은 난자성숙 과정의 조절 기작을 규명하기 위하여 생쥐의 미성숙 난자와 성숙난자에서 차이 나게 발현하는 유전자의 목록을 얻은 바 있다. 이들 유전자 중에서 Bcl-2 homolog인 Diva 유전자가 난자에 특이적으로 발현함을 본 연구를 통해 규명하였는데 이러한 Diva의 기능을 밝혀내기 위하여 immunoprecipitation (IP)과 Mass Spectrometry (MS)를 이용하여 Diva와 결합하여 상호작용하는 단백질을 동정하고자 하였다. 연구방법: NIH/3T3 세포주에 Diva를 encoding하는 pCMV-FLAG-Diva를 24 시간 동안 과발현 시키고 대조군으로는 유전자 없는 pCMV-FLAG empty vector를 transfection 하였다. FLAG에 특이적인 항체로 IP하여 Diva와 결합하는 면역복합체를 형성하게 한 후 이를 12% SDS-polyacrylamide gel 상에서 전기 영동하였고 Coomassie Blue 염색을 통해 단백질 발현양상을 관찰하였다. 대조군에서는 관찰되지 않으면서 Diva 유전자가 발현하는 실험군에서만 확인되는 밴드를 오려내어 trypsin을 사용하여 in-gel digestion 한 후 MS 분석을 시행하였다. 모든 mass spectra는 4700 Proteomics Analyzer (Applied Biosystems, Framingham, MA)에 의해 positive reflector mode에서 얻어졌다. 이렇게 얻어진 단백질들은 MASCOT Peptide Mass Fingerprint software (Matrixscience, London)을 이용하여 NCBI nonredundant database를 찾아서 동정하였다. 결과: Diva를 과발현하는 세포주에서만 관찰되는 15개 밴드에 대한 MS/MS 분석 결과, Diva와 결합하는 단백질로서 actin과 그 외에 ${\alpha}$-actinin, tropomyosin, tropomodulin 3 등의 actin-binding 단백질을 동정하였다. Diva를 과발현하는 NIH/3T3 세포주에서 면역 복합체를 형성하는 actin과 tropomyosin이 실제 난소 조직에서도 Diva와 결합하는지 IP와 Western blot을 통해 확인한 결과, actin과 tropomyosin 모두 Diva와 결합함을 확인함으로써, Diva는 이 두 단백질과 난소에서 상호작용함을 알 수 있었다. 결론: 본 연구는 Diva와 결합하여 상호작용하는 단백질들이 cytoskeletal system의 actin filament와 관계 있음을 규명한 최초의 보고이다. Diva가 actin과 tropomyosin과 결합하는 것을 고려해볼때, 난자 특이적인 Diva는 아마도 난자성숙 동안에 cytoskeletal system 을 조절하는 역할을 할 것으로 사료된다.