• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 춘계학술발표회
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• pp.67-67
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• 2018

Trichoderma species are a rich source of metabolites, but less known for biomedical potential. This work deals with antibacterial and antioxidant potentials of intracellular non-cytotoxic metabolites, extracted from Trichoderma atroviride (KNUP001). A total of 53 fractions was collected by column chromatography and tested for cytotoxicity by MTT assay. Only one fraction (F41) was found to be non-toxic to Vero cells with $95.4{\pm}0.61%$ of survival. The F41 was then subjected to chemical analysis, antibacterial and antioxidant assays. The F41 at $500{\mu}g.ml^{-1}$ showed the total antioxidant of $48.70{\pm}2.90%$, DPPH radical scavenging activity of $37.25{\pm}2.25$, nitric oxide (NO) radical scavenging activity of $54.55{\pm}1.95$ and $H_2O_2$ radical scavenging activity of $43.75{\pm}3.21$. The F41 at $25{\mu}g.ml^{-1}$ displayed antibacterial activity against E. coli ($14.25{\pm}0.2mm$), P. mirabilis ($10.4{\pm}0.6mm$), S. dysenteriae ($18.6{\pm}03mm$), S. paratyphi A ($14.1{\pm}1.1mm$), E. aerogenes ($5.6{\pm}0.4mm$) and S. marcescens ($14.25{\pm}0.2mm$). GC-MS analysis revealed the dominant presence of oleic acid C 18.1 (63.18%), n-hexadecanoic acid (6.17%), and ethyl oleate (4.93%) and potent molecules such as 8-[(2E)-2-(3-hydroxybenzylidene)hydrazinyl]-1,3,7-trimethyl-3,7-dihydro-1H-purine-2,6-dione, 2-(Dimethylamino)ethyl (1Z)-N-hydroxy-2-(4-morpholinyl)-2-oxoethanimidothioate, Fluorene in the F41, and virtual study revealed that these molecules are likely responsible for the antibacterial activities of F41. Hence, further investigation deserves on purification and characterization of the active metabolites from T. atroviride strain KNUP001 towards developing molecular leads to effective antibacterial drugs, and non-toxic to host cells.

• International Journal of Oral Biology
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• 제34권1호
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• pp.7-14
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• 2009

Nitric oxide (NO) acts as an intracellular messenger at the physiological level but can be cytotoxic at high concentrations. The cells within periodontal tissues, such as gingival and periodontal fibroblasts, contain nitric oxide syntheses and produce high concentrations of NO when exposed to bacterial lipopolysaccharides and cytokines. However, the cellular mechanisms underlying NO-induced cytotoxicity in periodontal tissues are unclear at present. In our current study, we examined the NO-induced cytotoxic mechanisms in human gingival fibroblasts (HGF). Cell viability and the levels of reactive oxygen species (ROS) were determined using a MTT assay and a fluorescent spectrometer, respectively. The morphological changes in the cells were examined by Diff-Quick staining. Expression of the Bcl-2 family and Fas was determined by RT-PCR or western blotting. The activity of caspase-3, -8 and -9 was assessed using a spectrophotometer. Sodium nitroprusside (SNP), a NO donor, decreased the cell viability of the HGF cells in a dose- and time-dependent manner. SNP enhanced the production of ROS, which was ameliorated by NAC, a free radical scavenger. ODQ, a soluble guanylate cyclase inhibitor, did not block the SNP-induced decrease in cell viability. SNP also caused apoptotic morphological changes, including cell shrinkage, chromatin condensation, and DNA fragmentation. The expression of Bax, a member of the proapoptotic Bcl-2 family, was upregulated in the SNP-treated HGF cells, whereas the expression of Bcl-2, a member of the anti-apoptotic Bcl-2 family, was downregulated. SNP augmented the release of cytochrome c from the mitochondria into the cytosol and enhanced the activity of caspase-8, -9, and -3. SNP also upregulated Fas, a component of the death receptor assembly. These results suggest that NO induces apoptosis in human gingival fibroblast via ROS and the Bcl-2 family through both mitochondrial- and death receptor-mediated pathways. Our data also indicate that the cyclic GMP pathway is not involved in NO-induced apoptosis.

• 한국식품영양과학회지
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• 제35권10호
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• pp.1291-1296
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• 2006

메밀 국수의 주 재료인 메밀분의 함량을 60%와 70%로 다르게 하여 건면, 압출면 그리고, 생면의 추출물을 조제하여 항돌연변이원성과 세포독성 효과를 조사하였다. 세포독성 효과를 알아보기 위해서 A549, Hep3B, MCF-7, AGS와 HeLa를 사용하였다. 이들 시료의 추출물의 암세포 성장 억제효과를 조사한 결과, 메밀 생면은 자궁암세포에서 68.5%의 높은 억제 활성을 나타내었으며 압출면의 경우는 위암세포에서 75.7%의 억제율을 나타내었다. 건면의 경우는 폐암 세포에서 80%의 높은 억제효과를 나타내었다. 그리고 돌연 변이성 및 항돌연변이 활성은 S. Typhimurium TA98과 TA100 균주를 이용한 돌연변이성 실험 결과 메밀면 자체에는 돌연변이성이 없었으며, 항돌연변이원성 실험에서는 직접변이원인 MNNG$(0.4{\mu}g/place)$의 경우 TA100 균주에서 시료농도 $160{\mu}g/place$에서 메밀분 70% 혼합분 생면 추출물의 경우 41%의 억제효과를 나타내었으며 4NQO$(0.15{\mu}g/place)$에 대해서는 같은 시료농도에서 TA98 균주에 대해서 43%의 억제 효과를 나타내었다. 그리고 메밀분 70% 혼합분 압출면의 경우 28%의 낮은 효과를 보였으나 S. Typhimurium TA98 균주에서는 같은 시료농도에서 45%의 억제효과를 나타내었으며 공시 시료 모두 농도 의존적으로 억제하는 것으로 나타났다.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제27권2호
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• pp.103-109
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• 2005

In spite of the ongoing advances, standard therapies for oral cancer still has some limitations in efficacy and in ability to prolong survival rate of advanced disease and result in significant functional defect and severe cosmetic deformity. Currently gene therapy using tumor suppressor gene is considered as a potent candidate for new therapeutic approaches that can improve efficacy and reduce complications. The purpose of this research is to identify the role of adenoviral vector to transfer HCCS-1 tumor suppressor gene in oral cancer cells and to find out whether there is a possibility for it to serve in the field of gene therapy. The human SCC-25 cell line was used for transfection. To determine the efficiency of the adenovirus as a gene delivery vector cell line was transduced with LacZ gene and analysed with X-gal staining. Northern blot was performed to confirm the tranfection with HSCC-1 gene and cell viability was assessed by cell cytotoxicity assay. We had successfully construct the recombinant HSCC-1 adenovirus(Ad5CMV-HCCS-1). DNA extracted from Ad5CMV-HCCS-1 revealed HCCS-1 gene is incorporated. The transduction efficiencies were over than 50% of SCC-25 cells with a MOI of 2 and over 95% with a MOI of 50. Northern blot analysis showed that a single 0.6kb mRNA transcript was expressed in Ad5CMV-HCCS-1 transduced SCC-25 cells. There was no or very low transcription HCCS-1 mRNA in wild and Ad5CMV-LacZ transduced SCC-25 cells. Cells transduced with Ad5CMV-HCCS-1 showed significant growth inhibition. By day 6, Ad5CMV-HCCS-1 treated cell count was decreased to 30% of mock-infected cells, while that of Ad5CMV-LacZ treated cells was 90% of mock-infected cells (p<0.05). Finally, these result suggest that the Ad5CMV-HCCS-1 has potential as a gene therapy tool for oral cancer.

• Journal of Ginseng Research
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• 제42권3호
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• pp.327-333
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• 2018

Background: Bioactive compounds in plant extracts are able to reduce metal ions to nanoparticles through the process of green synthesis. Panax ginseng is an oriental medicinal herb and an adaptogen which has been historically used to cure various diseases. In addition, the P. ginseng leaves-mediated gold nanoparticles are the value-added novel materials. Its potential as a cosmetic ingredient is still unexplored. The aim of this study was to evaluate the antioxidant, moisture retention and whitening properties of gold nanoparticles (PgAuNPs) in cosmetic applications. Methods: Cell-free experiments were performed to evaluate PgAuNP's antioxidant and moisture retention properties and inhibition activity on mushroom tyrosinase. Furthermore, in vitro cell cytotoxicity was evaluated using normal human dermal fibroblast and murine B16BL6 melanoma cells (B16) after treatment with increasing concentrations of PgAuNPs for 24 h, 48 h, and 72 h. Finally, in vitro cell assays on B16 cells were performed to evaluate the whitening effect of PgAuNPs through reduction of cellular melanin content and tyrosinase activity. Results: In vitro DPPH radical scavenging assay results revealed that PgAuNPs exhibited antioxidant activity in a dose-dependent manner. PgAuNPs exhibited moisture retention capacity and effectively inhibited mushroom tyrosinase. In addition, 3-(4,5-dimethyl-thiazol-2yl)-2,5-diphenyl tetrazolium bromide results revealed that PgAuNPs were not toxic to human dermal fibroblast and B16 cells; in addition, they significantly reduced melanin content, tyrosinase activity, and mRNA expression of melanogenesis-associated transcription factor and tyrosinase in B16 cells. Conclusion: Our study is the first report to provide evidence supporting that P. ginseng leaves-capped gold nanoparticles could be used as multifunctional ingredients in cosmetics.

• Archives of Pharmacal Research
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• 제21권5호
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• pp.581-590
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• 1998

We developed a novel water-soluble camptothecin analobue, CKD602, and evaluated the inhibition of topoisomerase I and the antitumor activities against mammalian tumor cells and human tumor xenografts. CKD602 was a nanomolar inhibitor of the topoisomerase I enzyme in the cleavable complex assay. CKD602 was found to be 3 times and slightly more potent than topotecan and camptothecin as inhibitors of topoisomerase, respecitively. In tumor cell cytotoxicity, CKD602 was more potent than topotecan in 14 out of 26 human cancer cell lines tested, while it was comparable to camptothecin. CKD602 was tested for the in vivo antitumor activity against the human tumor xenograft models. CKD602 was able to imduce regression of established HT-29, WIDR and CX-1 colon tumors, LX-1 lung tumor, MX-1 breast tumor and SKOV-3 ovarian tumor as much as 80, 94, 76, 67, 87% and 88%, respectively, with comparable body weight changes to those of topotecan. Also the therapeutic margin (R/Emax: maximum tolerance dose/$ED-{58}$) of CKD602 was significantly higher than that of topotecan by 4 times. Efficacy was determined at the maximal tolerated dose levels using schedule dependent i.p. administration in mice bearing L1210 leukemia. On a Q4dx4 (every 4 day for 4 doses) schedule, the maximum tolerated dose (MTD) was 25 mg/kg per administration, which caused great weight loss and lethality in <5% tumor bearing mouse. this schedule brought significant increase in life span (ILS), 212%, with 33% of long-term survivals. The ex vivo antitumor activity of CKD602 was compared with that of topotecan and the mean antitumor index (ATI) values recorded for CKD602 were significantly higher than that noted for topotecan. From these results, CKD602 warrants further clinical investigations as a potent inhibitor of topoisomerase I.

• 대한화학회지
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• 제57권5호
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• pp.634-639
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• 2013

암이나 혈관질환을 진단하기 위하여 영상의학기기들의 사용이 증가되고 있다. 영상의학기기 중 컴퓨터단층촬영(CT)은 가장 널리 이용되는 방법의 하나로서, CT를 하기 위해서는 조영제를 투여하여야 한다. 따라서, 본 연구에서는 리포솜의 표면에 금 입자를 코팅함으로써 조영제로 개발하고자 하였다. 금 입자를 코팅하기 위하여, 양이온성 리포솜을 제조하였고, 양이온성 리포솜의 표면에 음이온을 띄는 $Au^-$가 정전기적으로 코팅이 되게 하였다. 금 코팅 리포솜(GCL)의 크기는 $154.8{\pm}9.2$ nm 이었고, 표면전하는 $27{\pm}3.2$ mV 이었다. 리포솜의 형태는 주사전자현미경과 투과전자현미경으로 확인하였다. 리포솜 표면의 금 코팅 효율은 18% 였으며, MTT 분석을 한 결과, 금 코팅 과정에 대한 세포독성은 없었다. 그리고, 제조된 금 입자 코팅 리포솜을 CT로 촬영했을 경우 우수한 조영 효과를 나타냈다. 따라서 본 연구에서 제조된 GCL은 CT 조영제로서 다양한 혈관질환에 적용이 가능할 것이다.

• Journal of Ginseng Research
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• 제41권3호
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• pp.330-338
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• 2017

Background: Ginsenoside Rh2 (G-Rh2) is a ginseng saponin that is widely investigated because of its remarkable antitumor activity. However, the molecular mechanism by which (20S) G-Rh2 triggers its functions and how target animals avoid its cytotoxic action remains largely unknown. Methods: Phage display was used to screen the human targets of (20S) G-Rh2. Fluorescence spectroscopy and UV-visible absorption spectroscopy were used to confirm the interaction of candidate target proteins and (20S) G-Rh2. Molecular docking was utilized to calculate the estimated free energy of binding and to structurally visualize their interactions. MTT assay and immunoblotting were used to assess whether human serum albumin (HSA), bovine serum albumin (BSA), and bovine serum can reduce the cytotoxic activity of (20S) G-Rh2 in HepG2 cells. Results: In phage display, (20S) G-Rh2-beads and (20R) G-Rh2-beads were combined with numerous kinds of phages, and a total of 111 different human complementary DNAs (cDNA) were identified, including HSA which had the highest rate. The binding constant and number of binding site in the interaction between (20S)-Rh2 and HSA were $3.5{\times}10^5M^{-1}$ and 1, and those in the interaction between (20S) G-Rh2 and BSA were $1.4{\times}10^5M^{-1}$ and 1. The quenching mechanism is static quenching. HSA, BSA and bovine serum significantly reduced the proapoptotic effect of (20S) G-Rh2. Conclusion: HSA and BSA interact with (20S) G-Rh2. Serum inhibited the activity of (20S) G-Rh2 mainly due to the interaction between (20S) G-Rh2 and serum albumin (SA). This study proposes that HSA may enhance (20S) G-Rh2 water solubility, and thus might be used as nanoparticles in the (20S) G-Rh2 delivery process.

• 한국식품과학회지
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• 제35권4호
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• pp.690-695
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• 2003

Phellinus ribis(찔레 영지버섯)는 약용버섯으로 일부 항암 및 관절염 치료에 사용되고 있으나 그 활성이 전혀 알려져 있지 않아 1차적으로 40% ethanol 추출물의 생리활성을 확인하였다. 항산화 활성에 있어 nitrosamine의 원인물질인 nitrite 제거활성은 pH 1.2에서 0.5 mg/mL 이하 농도에서는 50% 이하, 그리고 1 mg/mL 처리 시 $64.0{\pm}1.6%$의 제거활성을 나타냈으며, 농도 의존적으로 증가하였다. DNA 손상 및 암, 당뇨, 간경화증, 심혈관질환 등의 원인물질에 관련하여 DPPH radicals의 제거활성은 0.5 mg/mL에서 $62.5{\pm}0.3%$, 1 mg/mL 처리 시 $91.3{\pm}0.8%$로 높게 나타났다. 2.5% linoleic acid의 자동산화에 미치는 항산화 활성을 측정한 결과 0.01 mg/mL 처리 시 p<0.05 수준에서, 0.5 및 1 mg/mL에서는 p<0.001 수준에서 유의성 있게 산화를 억제하였으며, superoxide dismutase 유사활성은 $530{\pm}81\;unit/g$으로 나타났다. 그리고, 항고혈압에 관련된 angiotensin converting enzyme 저해 활성은 $12.0{\pm}1.5%$로 그 활성이 미약하였다. Human 유래 암세포에 대한 세포독성 실험결과 폐암세포인 A549의 경우 100 mg/mL 농도에서도 16%로 나타나 세포독성이 미약하였고, 자궁암 세포인 HeLa의 경우 50 mg/mL에서 45%의 효과를, 위암 세표인 AGS에서는 5 mg/mL 처리 시 24%, 50 mg/mL에서는 76%, 그리고 간암 세포주인 SK-Hep-1의 경우 50 mg/mL에서 42%의 세포독성을 나타내었다. Salmonella typhimurium TA98 및 TA100에 대한 돌연변이원성 시험결과 찔레 영지버섯 추출물은 자연복귀 집락보다 histidine revertant colony 수가 2배 이상 증가하지 않았고, 추출물의 첨가농도를 증가시켜도 histidine revertant colony 수가 증가하지 않아 돌연변이성이 없었다.

• 동의생리병리학회지
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• 제23권6호
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• pp.1341-1348
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• 2009

Hoechunyanggyeok-san (HYS) is a traditional oriental herbal medicine widely used for treating inflammatory disorders. Although there are numerous clinical results of HYS reported in the literature of oriental hebal medicine, it has been rarely conducted to evaluate the immuno-biological activity. The present study was conducted to examine the anti-inflammatory effects of HYS extract (HYSE) in vivo and in vitro. To determine the cytotoxic concentration of HYSE, cell viability was tested by MTT assay. All four doses of HYSE (0.01, 0.03, 0.10 and 0.30 mg/ml) had no significant cytotoxicity during the entire experimental period. In order to measure NO levels in culture medium, the cells were treated with $1\;{\mu}g/ml$ of LPS 1h before adding HYSE for 24 h and then culture medium were reacted with Griess reagent. Increased NO production and iNOS expression were detected in LPS-activated cells compared to control. However, these increases were dose-dependently attenuated by treatment with HYSE. LPS plays a key role in leading to the massive production of pro-inflammatory cytokines such as TNF-$\alpha$, IL-$1{\beta}$ and IL-6 in macrophages. Thus, we next determined the levels of these cytokines. HYSE reduced the elevated production of TNF-$\alpha$, IL-$1{\beta}$ and IL-6 by LPS. Moreover, the effects of HYSE were in a dose-dependent manner. In vivo, histopathological study, HYSE effectively inheefed the increases of hind paw skin thicknesses and inflammatory cell infiltrations induced by carrageenan treatment. It, therefore, considered that HYSE will be favorably inheefed the acute edematous inanner. In s. These findings showed that HYSE could have anti-inflammatory effects through the reduction of NO and inflammatory cytokines in macrophage. Furthermore, the reduction of carrageenan-induced paw oedema by HYSE helps to understand its actions on inflammatory conditions.