• 한국수정란이식학회지
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• 제28권2호
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• pp.157-162
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• 2013

Methoxychlor (MXC) was developed to be a replacement for the banned pesticide DDT. HPTE [2,2-bis (p-hydroxyphenyl)-1,1,1-trichloroethane], which is an in vivo metabolite of MXC, has strong oestrogenic and anti-androgenic effects. MXC and HPTE are thought to produce potentially adverse effects by acting through oestrogen and androgen receptors. Of the two, HPTE binds to sex-steroid receptors with greater affinity, and it inhibits testosterone biosynthesis in Leydig cells by inhibiting cholesterol side-chain cleavage enzyme activity and cholesterol utilisation. In a previous study, MXC was shown to induce Leydig cell apoptosis by decreasing testosterone concentrations. I focused on the effects of MXC on male mice that resulted from interactions with sex-steroid hormone receptors. Sex-steroid hormones affect other organs including the kidney and liver. Accordingly, I hypothesised that MXC can act through sex-steroid receptors to produce adverse effects on the testis, kidney and liver, and I designed our experiments to confirm the different effects of MXC exposure on the male reproductive system, kidney and liver. In these experiments, I used pre-pubescent ICR mice; the puberty period in ICR mice is from postnatal day (PND) 45 to PND60. I treated the experimental group with 0, 100, 200, 400 mg MXC/kg b.w. delivered by an intra-peritoneal injection with sesame oil used as vehicle for 4 weeks. At the end of the experiment, the mice were sacrificed under anaesthesia. The testes and accessory reproductive organs were collected, weighed and prepared for histological investigation. I performed a chemiluminescence immune assay to observe the serum levels of testosterone, LH and FSH. Blood biochemical determination was also performed to check for other effects. There were no significant differences in our histological observations or relative organ weights. Serum testosterone levels were decreased in a dose-dependent manner; a greater dose resulted in the production of less testosterone. Compared to the control group, testosterone concentrations differed in the 200 and 400 mg/kg dosage groups. In conclusion, I observed markedly negative effects of MXC exposure on testosterone concentrations in pre-pubescent male mice. From our biochemical determinations, I observed some changes that indicate renal and hepatic failure. Together, these data suggest that MXC produces adverse effects on the reproductive system, kidney and liver.

• Journal of Microbiology and Biotechnology
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• 제19권3호
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• pp.331-337
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• 2009

Interleukin-13 (IL-13) has been proposed as a therapeutic target for bronchial asthma as it plays crucial roles in the pathogenesis of the disease. We developed an in vitro test system measuring transcriptional downregulatory activities on IL-13 as a primary screening method to select drug candidates from natural products. The promoter region of IL-13 (-2,048 to +1) was cloned into the upstream of a luciferase gene in the plasmid pGL4.14 containing the hygromycin resistance gene as a selection marker, generating pGL4.14-IL-13. The EL-4 thymoma and RBL-2H3 mast cells transiently expressing this plasmid highly produced the luciferase activities by responding to PI (PMA and ionomycin) stimulation up to 8-fold and 13-fold compared with the control, respectively, whereas cyclosporin A, a well-known antiasthmatic agent, significantly downregulated the activities. The BF1 clone of RBL-2H3 cells constitutively expressing pGL4.14-IL-13 was established by selecting surviving cells under a constant lethal dose of hygromycin treatment. The feasibility of this system was evaluated by measuring the downregulatory activities of 354 natural products on the IL-13 promoter using the BF1 clone. An extract from Morus bombycis (named TBRC 156) significantly inhibited PI-induced luciferase activities and IL-13 mRNA expression, but not the protein expression. Fisetin (named TBRC 353) inhibited not only PI-induced luciferase activities and mRNA expression, but also the IL-13 protein secretion, whereas myricetin (named TBRC 354) could not suppress the IL-13 expression at all. Our data indicated that this in vitro test system is able to discriminate the effects on IL-13 expression, and furthermore, that it might be suitable as a simple and time-saving primary screening system to select antiasthmatic agents by measuring transcriptional activities of the IL-13 promoter.

• Journal of Microbiology and Biotechnology
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• 제25권7호
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• pp.1170-1176
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• 2015

Ginsenosides, the major active component of ginseng, are traditionally used to treat various diseases, including cancer, inflammation, and obesity. Among these, compound K (CK), an intestinal bacterial metabolite of the ginsenosides Rb₁, Rb₂, and Rc from Bacteroides JY-6, is reported to inhibit cancer cell growth by inducing cell-cycle arrest or cell death, including apoptosis and necrosis. However, the precise effect of CK on breast cancer cells remains unclear. MCF-7 cells were treated with CK ($0-70{\mu}M$) for 24 or 48 h. Cell proliferation and death were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and flow cytometry assays, respectively. Changes in downstream signaling molecules involved in cell death, including glycogen synthase kinase $3\beta$ ($GSK3\beta$), $GSK3\beta$, $\beta$-catenin, and cyclin D1, were analyzed by western blot assay. To block $GSK3\beta$ signaling, MCF-7 cells were pretreated with $GSK3\beta$ inhibitors 1 h prior to CK treatment. Cell death and the expression of $\beta$-catenin and cyclin D1 were then examined. CK dose- and time-dependently inhibited MCF-7 cell proliferation. Interestingly, CK induced programmed necrosis, but not apoptosis, via the $GSK3\beta$ signaling pathway in MCF-7 cells. CK inhibited $GSK3\beta$ phosphorylation, thereby suppressing the expression of $\beta$-catenin and cyclin D1. Our results suggest that CK induces programmed necrosis in MCF-7 breast cancer cells via the $GSK3\beta$ signaling pathway.

• 대한약침학회지
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• 제19권1호
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• pp.37-44
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• 2016

Objectives: This study examined the relative efficacies of a derivative of betulinic acid (dBA) and its poly (lactide-co-glycolide) (PLGA) nano-encapsulated form in A549 lung cancer cells in vivo and in co-mutagen [sodium arsenite (SA) + benzo[a]pyrene (BaP)]-induced lung cancer in mice in vivo. Methods: dBA was loaded with PLGA nanoparticles by using the standard solvent displacement method. The sizes and morphologies of nano-dBA (NdBA) were determined by using transmission electron microscopy (TEM), and their intracellular localization was verified by using confocal microscopy. The binding and interaction of NdBA with calf thymus deoxyribonucleic acid (CT-DNA) as a target were analyzed by using conventional circular dichroism (CD) and melting temperature (Tm) profile data. Apoptotic signalling cascades in vitro and in vivo were studied by using an enzyme-linked immunosorbent assay (ELISA); the ability of NdBA to cross the blood-brain barrier (BBB) was also examined. The stage of cell cycle arrest was confirmed by using a fluorescence-activated cell-sorting (FACS) data analysis. Results: The average size of the nanoparticles was ~ 110 nm. Confocal microscopy images confirmed the presence of NdBA in the cellular cytoplasm. The bio-physical properties of dBA and NdBA ascertained from the CD and the Tm profiles revealed that NdBA had greater interaction with the target DNA than dBA did. Both dBA and NdBA arrested cell proliferation at G0/G1, NdBA showing the greater effect. NdBA also induced a greater degree of cytotoxicity in A549 cells, but it had an insignificant cytotoxic effect in normal L6 cells. The results of flow cytometric, cytogenetial and histopathological studies in mice revealed that NdBA caused less nuclear condensation and DNA damage than dBA did. TEM images showed the presence of NdBA in brain samples of NdBA fed mice, indicating its ability to cross the BBB. Conclusion: Thus, compared to dBA, NdBA appears to have greater chemoprotective potential against lung cancer.

• 한국응용과학기술학회지
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• 제36권4호
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• pp.1303-1311
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• 2019

본 연구는 열수와 에탄올 용매에 따른 복령 및 산수유 추출물의 농도별로 항산화 활성, 항염증 효과 및 신경세포 보호효과에 미치는 영향을 살펴보고자 진행하였다. 추출물들의 총 polyphenol 함량은 복령 열수 추출물에서 가장 높았고, 복령 에탄올 추출물, 산수유 열수 추출물, 산수유 에탄올 추출물 순으로 유의적으로 감소되는 경향을 나타냈다. DPPH 및 ABTS radical 소거 활성능은 추출물의 농도 의존적으로 증가하였고 산수유 열수 추출물은 모든 추출물에서 항산화 활성이 가장 높게 나타났으며, 복령 열수 추출물은 에탄올 추출물에 비해 높은 활성이 확인되었다. 복령과 산수유에 함유된 총 polyphenol과 항산화 성분을 추출하기 위해서는 에탄올 추출방법 보다는 열수 추출방법이 효과적인 방법이라고 사료된다. LPS로 염증이 유도된 RAW 264.7 세포에서 복령추출물이 산수유 추출물에 비해 NO 생성 억제 효과가 우수한 것으로 확인되었다. MPP⁺에 의해 유도된 SH-SY5Y 신경세포에 복령 열수 추출물은 산수유 열수 추출물에 비해 산화적 스트레스로부터 신경세포 보호효과가 우수하게 나타났다. 복령과 산수유 추출방법에 따른 항산화, 항염 및 산화적 스트레스로부터의 신경세포 보호효과를 갖는 기능성 소재로서 가능성이 있다고 사료된다.

• 한국산학기술학회논문지
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• 제17권11호
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• pp.186-195
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• 2016

동해 울릉도-독도 2정점에서 식물플랑크톤의 군집조성 및 수직적 영양염분포 특성과 표층 현장수를 이용하여 N, P, NP, Fe을 첨가하여 식물플랑크톤의 영양염소비 및 성장특성을 파악하였다. 현장에서 영양염류농도는 유광층 상부에서 낮았고, 100 m보다 깊은 수심에서 증가하였다. N:P비는 Redfield비(16)보다 표층에서 21-25로 높았고, 조사 최대심도인 400 m 층에서는 6 전후로 극히 낮게 관찰되었다. 식물플랑크톤의 총개체수는 St.UD3과 St.50에서 각각 $4.9{\times}10^5cells\;L^{-1}$과 $1.9{\times}10^5cells\;L^{-1}$로 관찰되었다. 주요 우점종은 St.UD3에서 침편모조류 Heterosigma akashiwo, 은편모조류 Crytomonas spp., 규조류 Leptocylindrus danicus로 파악되었고, St.50에서는 규조류 Chaetoceros socialis, H. akashiwo, L. danicus로 각각 관찰되었다. 영양염첨가실험의 +N과 +NP실험군에서 빠른 식물플랑크톤의 성장반응을 보였고, +P실험군에서는 대조군과 유사하게 식물플랑크톤 성장이 관찰되지 않았다. 아울러 Fe을 첨가한 대조군, +N, +P, +NP실험군에서 Fe을 첨가하지 않은 다른 실험군과 비교하여 형광값의 유의한 차이를 보이지 않았다는 것은 Fe첨가에 따른 식물플랑크톤의 성장반응이 명확하지 않다는 것을 의미한다(p>0.05). 결과적으로 현장에서 우점한 H. akashiwo 와 L. danicus는 영양염이 추가적으로 공급된 환경의 생물검증실험에서도 빠르게 반응하여 높은 개체수로 우점한 것을 파악하였고, 특히 +Fe 실험군에서 L. danicus의 빠른 성장이 두드려졌다.

• 대한화장품학회지
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• 제33권4호
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• pp.269-273
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• 2007

산업발전에 따라 환경분야가 대두됨에 따라 민감성 피부에 화장품을 사용하는데 있어 부작용이 증가하였다. 이에 따라 저자극성 방부제의 개발을 위하여 항암효과가 입증된 침엽수 중 구상나무, 향나무의 활성성분을 추출하여 항균력을 측정 평가하였다. 항균력 측정에는 Broth dilusion법이 사용되었으며, 균주는 그람 음성균인 Escherichia coli와 그람 양성균인 Staphylococcus aureus를 사용하였다. 그 결과 항균효과는 향나무와 구상나무가 625 ppm에서 methyl paraben에 비하여 각각 17.02 %, 8.5 % 더 높은 효과를 나타냈다. 항산화 효과 측정은 DPPH법, 총 폴리페놀 함량은 Folin-Denis법, 총 플라보노이드의 함량은 Nieva Moreno법을 사용하여 분석하였다. 항산화 효과에서 향나무는 5,000 ppm에서 45 %, 구상나무는 44 %의 항산화 효과를 나타냈으며 분석 결과 총 폴리페놀과 총 플라보노이드 함유량과 비례하였다. 침엽수 추출물의 세포독성실험은 피부섬유아세포인 CCK-986sk를 사용하였다. 그 결과 향나무, 구상나무가 39 ppm에서 1,250 ppm까지의 모든 농도에서 95% 이상의 높은 세포 생존률을 나타냈다. 이상을 종합해 본 결과 향나무 줄기, 구상나무 잎은 합성물질을 대체할 수 있는 방부제 및 보조 항산화제로서 사용이 가능하다고 할 수 있다.

• Journal of Ginseng Research
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• 제44권6호
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• pp.799-807
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• 2020

Background: The skin acts as a barrier to protect organisms against harmful exogenous agents. Compound K (CK) is an active metabolite of ginsenoside Rb1, Rb2 and Rc, and researchers have focused on its skin protective efficacy. In this study, we hypothesized that increased expression of the serine protease inhibitor Kazal type-5 (SPINK5) may improve skin barrier function. Methods: We screened several ginsenosides to increase SPINK5 gene promoter activity using a transactivation assay and found that CK can increase SPINK5 expression. To investigate the protective effect of CK on the skin barrier, RT-PCR and Western blotting were performed to investigate the expression levels of SPINK5, kallikrein 5 (KLK5), KLK7 and PAR2 in UVB-irradiated HaCaT cells. Measurement of transepidermal water loss (TEWL) and histological changes associated with the skin barrier were performed in a UVB-irradiated mouse model and a 1-chloro-2,4-dinitrobenzene (DNCB)-induced atopic dermatitis-like model. Results: CK treatment increased the expression of SPINK5 and decreased the expression of its downstream genes, such as KLKs and PAR2. In the UVB-irradiated mouse model and the DNCB-induced atopic dermatitis model, CK restored increased TEWL and decreased hydration and epidermal hyperplasia. In addition, CK normalized the reduced SPINK5 expression caused by UVB or DNCB, thereby restoring the expression of the proteins involved in desquamation to a level similar to normal. Conclusions: Our data showed that CK contributes to improving skin-barrier function in UVB-irradiated and DNCB-induced atopic dermatitis-like models through SPINK5. These results suggest that therapeutic attempts with CK might be useful in treating barrier-disrupted diseases.

• 한국식품위생안전성학회지
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• 제19권4호
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• pp.171-175
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• 2004

국내에서 유통되는 식용 타르색소 9종에 대하여 한국, 일본, 미국 및 JECFA의 색소시험법에 따라 비교${\cdot}$시험하였다. 식용색소의 물불용물 함량은 각 기관의 시험법에서 차이가 없었으며, 모두 규격기준에 적합하였다. 식용색소 녹색제3호, 적색제3호, 청색제2호 및 황색제4호의 염화물 및 황산염 함량은 각 기관의 방법에 따라 약간 달랐지만 모두 규격기준에 적합하였고, JECFA와 미국방법에서는 분석하는데 시간이 더 많이 소요되었다. 비소 함량은 한국과 일본에서 비색법으로 비교하였으며, 규격기준이 미국 및 JECF와 달랐다. 중금속 함량은 모두 규격기준에 적합하였으나 한국은 비색법, 일본은 원자흡광도법, 미국과 JECFA는 두 가지 방법을 모두 사용하였다. 비술폰화방향족제 1급 아민의 함량은 분석시료 모두 $0.0005\%$(aniline으로서)이하로서 규격기준($0.01\%$이하)에 모두 적합하였다.

• 한국식품과학회지
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• 제40권6호
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• pp.696-701
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• 2008

이 연구의 목적은 더덕 및 도라지 에틸아세테이트 분획물(CLEA나 PGEA)이 다양한 항산화 시스템을 가동시켜 sodium nitroprusside(SNP)에 의해 유도된 산화적 스트레스에 대항하여 세포를 보호해 줄 수 있을지 연구하는 것이다. 0.5 mM의 SNP를 HepG2 세포주에 24시간 동안 노출시켰을 때 세포 생존율이 감소하였으나, CLEA와 PGEA 천처리에 의해 SNP 노출에 의한 세포 생존율 감소가 저해되었다. 또한 항산화 시스템들 발현에 미치는 CLEA와 PGEA의 영향을 RT-PCR로 알아보았다. Catalase, glucose-6-phosphate dehydrogenase(G6PD) 그리고 metallothionein (MT)-1A mRNA 수준이 CLEA를 세포에 24시간 동안 처리한 후 증가하였고, Mn superoxide dismutase, catalase, G6PD, MT-1A 와 MT-2A mRNA 수준이 PGEA 처리에 의해 증가하였다. 우리는 CLEA와 PGEA가 아마 다양한 항산화 시스템들을 증가시키는 간접적인 항산화 효과를 나타내며, 이들 효과들이 산화적 스트레스로부터 세포를 보호해 줄 것이라 추정하였다.