• 한국임상수의학회지
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• 제28권6호
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• pp.549-554
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• 2011

FK506은 말기 간암환자의 간이식 후 널리 사용되는 면역억제제이다. Dexamethasone은 세포독성 암 치료에서 오심 방지, 정상세포의 보호와 기타 이유 등의로 빈번하게 병용처치된다. 본 연구의 목적은 간암세포주(Hep3B)에서 FK506의 항암효과와 FK506에 의한 항암효과에 대한 dexamethasone의 억제효과를 알아보기 위함이다. 세포의 손상은 세포 생존성 평가와 LDH 및 세포내 ROS 양의 측정으로 평가 하였다. 세포내 칼슘 농도([$Ca^{2+}$]i)와 JNK, Bax 단백질의 발현 정도도 평가하였다. FK506의 처치는 Hep3B의 세포사를 유도하였으며 세포생존성의 감소와 LDH, ROS 및 [$Ca^{2+}$]i 를 증가시켰다. FK506은 Bax와 JNK 의 활성을 증가시켰으며 Bcl-2의 활성을 억제하였다. Dexamethasone 처치 그 자체는 세포생존성, LDH와 ROS에 영향을 주지 않았다. 그러나 dexamethasone과 FK506의 병용처치는 FK506에 의한 LDH 방출, ROS 생성 및 JNK의 활성을 감소시켰다. 이 결과는 간암세포주에서 FK506은 항암효과를 가지지만 dexamethasone의 병용처치는 FK506에 의한 항암효과를 길항한다.

• 대한수의학회지
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• 제54권4호
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• pp.209-218
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• 2014

Experimental autoimmune encephalomyelitis (EAE), an animal model of human multiple sclerosis (MS), reflects pathophysiologic steps in MS such as the influence of T cells and antibodies reactive to the myelin sheath, and the cytotoxic effect of cytokines. Galectin-9 (Gal-9) is a member of animal lectins that plays an essential role in various biological functions. The expression of Gal-9 is significantly enhanced in MS lesions; however, its role in autoimmune disease has not been fully elucidated. To identify the role of Gal-9 in EAE, we measured changes in mRNA and protein expression of Gal-9 as EAE progressed. Expression increased with disease progression, with a sharp rise occurring at its peak. Gal-9 immunoreactivity was mainly expressed in astrocytes and microglia of the central nervous system (CNS) and macrophages of spleen. Flow cytometric analysis revealed that $Gal-9^+CD11b^+$ cells were dramatically increased in the spleen at the peak of disease. Increased expression of tumor necrosis factor (TNF)-R1 and p-Jun N-terminal kinase (JNK) was observed in the CNS of EAE mice, suggesting that TNF-R1 and p-JNK might be key regulators contributing to the expression of Gal-9 during EAE. These results suggest that identification of the relationship between Gal-9 and EAE progression is critical for better understanding Gal-9 biology in autoimmune disease.

• 폴리머
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• 제27권3호
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• pp.217-225
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• 2003

약물의 방출 경향을 제어할 목적으로 폴리비닐피롤리돈 (PVP) 또는 염화나트륨을 함유한 항암제 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU, carmustine)/poly(L-lactide-co-glycolide) (PLGA) 웨이퍼를 직접 압축성형 방법에 의해 직경 3 mm와 두께 1 mm의 조건으로 제조하였다. 생체외 방출실험에서 BCNLT/PLGA 웨이퍼로부터 약물 방출거동은 웨이퍼에 함유된 첨가제에 의해 조절할 수 있었다. BCNU의 초기 방출량과 방출속도 및 기간은 염화나트륨 또는 PVP의 첨가에 외해 변화하였다. 9L gliosarcoma세포를 이용한 생체 외 항암 활성 실험에서 첨가제가 함유된 웨이퍼로부터 방출된 BCNU를 처리한 세포의 생존율을 분석하여 평가했고, 그 결과 지속적으로 9L gliosarcoma세포의 성장을 억제함을 확인하였다. 9L gliosarcoma세포에 대한 첨가제 함유 BCNU/PLGA 웨이퍼로부터 방출된 BCNU 약효 검색을 통하여 웨이퍼로부터 BCNU가 지속적으로 방출되어 9L gliosarcoma세포의 생존과 증식을 효과적으로 억제함을 확인하였다. 특히, 첨가제 함유 BCNU/PLGA 웨이퍼의 지속적인 성장 억제는 첨가제를 함유하지 않은 웨이퍼의 것보다 더욱 효과적이었다. 또한 염화나트륨 함유 BCNU/PLGA 웨이퍼가 PVP 함유 BCNU/PLCA 웨이퍼보다 세포 증식 억제 효과가 뛰어남을 보였다.

• 한국식품위생안전성학회지
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• 제15권3호
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• pp.241-247
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• 2000

국내 유통식품에서 verotoxin(VT)을 생성하는 대장균의 오염도를 조사하고 이로 인한 식중독의 발생을 사전에 예방하기 위하여 1997년부터 1999년 사이 햄버거. 식육 및 채소류에서 VT생성 대장균의 분포조사를 실시하였으며, 이 결과 분리된 대장균에 대한 분자생물학적 특성을 조사하였다 식육, 햄버거 및 채소류 총 1,700건의 식품 중에서 VT를 생성하는 대장균이 검출된 것은 3건으로 매우 낮은 검출율을 보였는데, 분리주의 혈청형은 각각 026 : H4, 0157 : H7 및 055 : Hl2였다. 혈청형 026 : H4 분리주는 VT I과 VT II를 생성하였고,055 : Hl2주는 VT I을 각각 생성하였으며, 두 분리주에서 eae유전자와 60 MDa plasmid DNA는 검출되지 않았다. 혈청형 0157 : H7주는 VT II를 생성하였고 60 MDa plasmid DNA는 검출되었으나 eae유전자는 확인되지 않았다. 따라서 분리된 VT생성 대장균들은 eae 유전자가 검출되지 않아 질병유발 가능성은 낮은 것으로 추정되었다. 항생제 내성능에서 혈청형 0157 : H7 분리주는 ampicillin과 streptomycin에 내성을 나타낸 반면, 혈청형 026 : H4주와 혈청형 055 : Hl2주는 ampicillin, carbenicillin, cephalothin, trimethoprim/sulfamethoxazole과 tetracycline에 내성을 보여 분리주의 다재내성능을 확인하였다 세포배양액으로 실시한 vero cell과 HeLa cell에 대한 세포독성능은 3주의 분리주에서 확인되었다.

• 한국근적외분광분석학회:학술대회논문집
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• 한국근적외분광분석학회 2001년도 NIR-2001
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• pp.4105-4105
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• 2001

Near infrared spectroscopy (NIRS) was employed to qualify and quantify on survival, the injury rate and apoptosis of the human breast cancer cell line MCF-7 cells. MCF-7 cells were cultured in RPMI medium supplemented with 10% FCS in a 95% air and 5% CO2 atmosphere at 37$^{\circ}C$. For the viable cells preparation, cells were de-touched by 0.1% of trypsin treatment and washed with RPMI supplemented with 10% FCS medium by centrifugation at 1000 rpm for 3min. For the dead cells preparation, cells were de-touched by a cell scraper. The cells were counted by a hemacytometer, and the viability was estimated by the exclusion method with frypan blue dye. Each viable and dead cells were suspended in PBS (phosphate bufferred saline) or milk at the cell density desired. For the quantitative determination of cell death by measuring the LDH (lactate dehydrogenase) activity liberated from cells with cell membrane injuries, LDH-Cytotoxic Test Wako (Wako, Pure Pharmaceutical Co. Ltd., Japan) was used. We found that NIRS measurement of MCF-7 cells at the density range could evaluate and monitor the different characteristics of living cells and dead cells. The spectral analysis was performed in two wavelength ranges and with 1,4, 10 mm pathlength. Different spectral data pretreatment and chemometrics methods were used. We applied SIMCA classificator on spectral data of living and dead cells and obtained good accuracy when identifying each class. Bigger variation in the spectra of living cells with different concentrations was observed when compared to the same concentrations of dead cells. PLS was used to measure the number of cells in PBS. The best model for measurement of dead cells, as well as living cells, was developed when raw spectra in the 600-1098 nm region and 4 mm pathlength were used. Smoothing and second derivative spectral data pretreatment gave worst results. The analysis of PLS loading explained this result with the scatter effect found in the raw spectra and increased with the number of cells. Calibration for cell count in the 1100-2500 nm region showed to be very inaccurate.

• 한국식품영양과학회지
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• 제26권2호
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• pp.254-260
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• 1997

본 연구에서는 in vitro와 in vivo에서 sarcoma-180 cells과 마우스를 이용하여 김치의 세포독성 효과와 면역계, 특히 대식세포의 탐식능의 활성 증강에 미치는 효과를 중심으로 김치의 항암기작을 연구하였다. 김치가 발효되기 전인 0주(생김치)와 최적의 숙성도를 나타내는 3주 김치$(5^{\circ}C)$를 메탄올, 헥산추출물과 MSF(methanol soluble fraction)로 분리 조제하여 실험에 이용하였다. 김치추출물은 탐식능 증진효과를 나타냈으며, 0주 생김치 보다 3주 발효 김치에서 더욱더 활성이 높았다. 이러한 실험결과를 요약하면 다음과 같다. 1. Sarcoma-180 종양세포에 대한 시료의 직접적인 작용에서는 김치추출물에 의하여 총 세포수가 크게 줄어드는 것을 볼 수 있었다. 3주 발효 김치추출물은 같은 농도에서 0주(생김치) 시료 보다 세포독성작용이 더 높았다. 2. Phagocytic activity는 in vitro와 in vivo에서 대조군에 비해 김치추출물의 모든 시료에서 탐식율이 높게 나타났다. In vitro에서는 대조군이 41.3%에 비해, 3주 김치의 메탄올추출물은 57.3%, 헥산추출물에서는 49.2%를 나타냈고, in vivo에서는 대조군이 24.3%에 비해, 3주 김치의 메탄올추출물은 57.0%, MSF에서는 55.0%였다. 3. In vitro에서는 메탄올추출물에서, in vivo에서는 3주 발효 김치의 메탄올추출물과 MSF에서 대식세포 당 평균 2개 이상의 C. albicans를 탐식하고 있음을 알 수 있었으며 0주(생김치) 보다 3주 김치 추출물에서 더욱더 탐식능 증진효과를 나타냈다.

• Journal of Acupuncture Research
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• 제23권3호
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• pp.177-190
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• 2006

Objectives : It has long been known about the anticancer effect of GRR-HAS, however, it has not been systemically determined the differentially regulated genes by GRR-HAS in cancer cells. The purpose of this study is to screen the GRR-HAS mediated differentially expressed genes in cancer cells such as HepG2 hepatoma cell lines. Oligonucleotide microarray and proteomic approaches were employed to screen the differential expression genes. Methods : GRR~HAS was prepared by boiling and stored at $-70^{\circ}C$ until use. Cells were treated with various concentrations of GRR-HAS (0.1, 0.5, 1.5, 10, $20mg/m{\ell}$) for 24 h. Cell toxicity was tested by MTT assay. To screen the differentially expressed genes in cancer cells, cells were treated with $1.5mg/m{\ell}$ of GRR-HAS. For oligonucleotide microarray assay, total RNA was used for gene expression analysis using oligonucleotide genechip (Human genome Ul33 Plus 2.0., Affimatrix Co.). For proteomic analysis, total protein was analyzed by 2D gel electrophoresis and Q-TOF mass spectrometer. Results : It has no cytotoxic effects on both HepG2 cells in all concentrations(0.1, 0.5, 1.5, 10,$20mg/m{\ell}$). In oligonucleotide microarray assay, the number of more than twofold differentially regulated known genes was 320 with 6 up-regulated and 314 down-regulated genes in HepG2 cells. In proteomic analysis, three spots were identified by 2D-gel electrophoresis and Q-TOF analysis. One down -regulated protein was protein disulfide isomerase and up-regulated proteins were fatty acid binding protein 1 and 14-3-3 gan1lTIa protein by $1.5mg/m{\ell}$ of CRR-HAS. Discussion : This study showed the comprehensive gene expression analysis using oligonucleotide microarray for the screening of GRR-HAS mediated differentially regulated genes. These results will provide a better application of GRR-HAS in cancer field and drug target development.

• 대한한의학방제학회지
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• 제22권1호
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• pp.151-165
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• 2014

Objectives : The aim of this study was to evaluate the efficacy of Aconiti Lateralis Preparata Radix (AP) and Acanthopanacis Cortex (AT) extracts in bone-derived adipocyte OP9 cell, osteoclast and osteoblast-like MG63 cells. Methods : MTT assay was used to evaluate the cytotoxicity of AP and AT extracts on OP9, osteoclast and MG63 cells. OP9 cells were treated with AP and AT, and the alterations in fat storage in the cells were determined by the Oil red O. To explain effects of RANKL-induced osteoclast differentiation in bone marrow macrophages, we performed the TRAP staining. The protein level of CAAAT/enhancer binding protein alpha ($C/EBP{\alpha}$) and peroxisome proliferator-activated receptor ${\gamma}$ ($PPAR{\gamma}$) as a adipocyte differentiation marker, and adiponectin was examined using western blot in differentiated OP9 cells. Effects of related genes were confirmed by luciferase assay using reporter assay. Results : AP and AT was not toxic on OP9 and MG63 cells, but AT was a little cytotoxic to osteoclast at the dose of $100{\mu}g/m{\ell}$. They could inhibit differentiation of OP9 cells and osteoclast with results of oil red O staining and TRAP staining. By western blot, AP and AT decreased the expression of $PPAR{\gamma}$ and $C/EBP{\alpha}$ which is the key transcription factor in adipogenesis and adiponectin secretion. AT also inhibited the BMP-4 activity in luciferase assay. AP also inhibited BMP-4 and Wnt3a activity, stimulated ER-${\beta}$ activity but inhibited androgen receptor activity. Conclusions : These results show AP and AT can be useful in osteoporosis and obesity via inhibition of osteoclast and adipocyte differentiation.

• Journal of Microbiology and Biotechnology
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• 제15권4호
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• pp.756-766
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• 2005

The signaling mechanism underlying aburatubolactam C-induced FasL upregulation was investigated in human Jurkat T cells. After treatment with aburatubolactam C, the src-family PTKs $p56^{lck}\;and\;p59^{fyn}$, and MAP kinases ERK2 and JNK1, were activated prior to FasL upregulation; Both $p56^{lck}\;and\;p59^{fyn}$ were directly activated 2.4- and 2.2-fold, respectively, in vitro by aburatubolactam C. The aburatubolactam C-induced cellular changes, including the activation of ERK2 and INK1, and FasL upregulation, were completely prevented by the PTK inhibitor genistein. The activation of protein kinase C (PKC$\delta,\;\epsilon\;and\;\mu$ was also induced following aburatubolactam C treatment. Although the activation of $p56^{lck}$ and tyrosine phosphorylation of the cellular proteins were not blocked by the PKC inhibitor GFl09203X, the activation of ERK2 was completely abrogated, along with a detectably enhanced JNK1 activation; FasL upregulation, and apoptosis. However, the FasL upregulation and apoptosis were significantly inhibited by the PKC activator PMA, with a remarkable increase in the ERK2 activation. The cytotoxic effect of aburatubolactam C was reduced in the presence of the anti-Fas neutralizing antibody ZB-4. Although ectopic expression of Bcl-2 failed to completely block the cytotoxicity of aburatubolactam C, it was clearly suppressed. The c-Fos mRNA expression was upregulated in a biphasic manner, where the second phasic expression overlapped with the FasL upregulation. Accordingly, these results demonstrate that aburatubolactam C-induced apoptosis is exerted, at least in part, by FasL upregulation dictated by activation of the PTK ($p56^{lck}\;and\;p59^{fyn}$) /JNKI pathway, which is negatively affected by the concurrent activation of the PKC/ERK2 pathway proximal to PTK activation.

• 한국미생물·생명공학회지
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• 제44권3호
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• pp.285-292
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• 2016

본 연구에서는 에탄올과 고압 열수 추출에 의해 준비된 벼세포 추출물의 창상 치유 효능 평가를 위해, 인간 섬유아세포와 각질세 포에 추출물 처리하여 세포 독성, 세포 이동능, collagen 단백질 및 유전자의 발현 양상에 대해 실험을 진행하였다. 그 결과 추출물은 처리한 범위에서 두 세포에 전혀 독성을 나타내지 않았으며, 대조구에 비해 증식을 최대 25% 촉진시키는 것으로 나타났다. 또한 인위적으로 세포에 창상을 낸 후 세포 이동능을 평가하였을 때, 비처리군에 비해 처리 농도에 비례하여 230−450% (섬유아세포), 170−350%(각질세포) 이동성이 향상되는 것을 확인할 수 있었다. 창상 치유의 중요한 역할을 하는 collagen의 유전자와 단백질 모두 추출물 처리시 농도의존적으로 증가하는 것을 확인할 수 있었다. 특히 에탄올 추출물 100 mg/ml 처리시에는 양성 대조군인 vitamin C (500 μM)와 유사한 효능을 나타내었다. 따라서 본 연구의 결과는 벼세포 추출물이 창상 치유제로서의 활용 가능성에 대해 제시하고 있다.