• Archives of Pharmacal Research
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• 제24권1호
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• pp.74-78
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• 2001

A preparation of water soluble components (EA) was made from carpophores of Elfvingia applanata (Pers.) Karst and its in vitro antiviral activity on vesicular stomatitis virus [(Indiana serotype, VSV(IND)] was investigated by plaque reduction assay. EA exhibited potent antiviral activity on VSV(IND) growth and negligible cytotoxicity on Vero cells, 50% effective concentration ($EC_{50}C$/) of 104$ug\textrm\/ml$ and 50% cytotoxic concentration ($CC_{50}C$) of 3,793$ug\textrm\/ml$, respectively. Selectivity index (Sl $CC_{50}C$/$EC_{50}C$) of EA on Vero cell and VSV(IND) was about 36.5. EA did not display either a direct virucidal effect on V5V(IND) or induction of antiviral substance by Vero cells upon its treatment. Thus, the mode of antiviral activity of EA was studied at steps of viral adsorption onto cell. When both EA and virus were added to cell monolayers, titer of cell-free virus in culture supernatant increased in ca. 30-40% compared with that of control group and titer of cell-associated virus was 60-100% higher than that of control group. These results suggested that antiviral activity of EA on VSV(IND) might be due to the hindrance of viral entry to cells at eITher endocytosis or loss of envelope.

• Nutrition Research and Practice
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• 제12권1호
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• pp.41-46
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• 2018

BACKGROUND/OBJECTIVES: Exposure of the normal lung tissue around the cancerous tumor during radiotherapy causes serious side effects such as pneumonitis and pulmonary fibrosis. Radioprotectors used during cancer radiotherapy could protect the patient from side effects induced by radiation injury of the normal tissue. Delphinidin has strong antioxidant properties, and it works as the driving force of a radioprotective effect by scavenging radiation-induced reactive oxygen species (ROS). However, no studies have been conducted on the radioprotective effect of delphinidin against high linear energy transfer radiation. Therefore, this study was undertaken to evaluate the radioprotective effects of delphinidin on human lung cells against a proton beam. MATERIALS/METHODS: Normal human lung cells (HEL 299 cells) were used for in vitro experiments. The 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay assessed the cytotoxicity of delphinidin and cell viability. The expression of radiation induced cellular ROS was measured by the 2'-7'-dicholordihydrofluorescein diacetate assay. Superoxide dismutase activity assay and catalase activity assay were used for evaluating the activity of corresponding enzymes. In addition, radioprotective effects on DNA damage-induced cellular apoptosis were evaluated by Western blot assay. RESULTS: Experimental analysis, including cell survival assay, MTT assay, and Western blot assay, revealed the radioprotective effects of delphinidin. These include restoring the activities of antioxidant enzymes of damaged cells, increase in the levels of pro-survival protein, and decrease of pro-apoptosis proteins. The results from different experiments were compatible with each to provide a substantial conclusion. CONCLUSION: Low concentration ($2.5{\mu}M/mL$) of delphinidin administration prior to radiation exposure was radioprotective against a low dose of proton beam exposure. Hence, delphinidin is a promising shielding agent against radiation, protecting the normal tissues around a cancerous tumor, which are unintentionally exposed to low doses of radiation during proton therapy.

• 한국자원식물학회지
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• 제27권3호
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• pp.215-222
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• 2014

This study was conducted to isolate a compound with anticancer properties from the roots of Peucedanum japonicum Thunb. (Umbelliferae), and to evaluate the efficacy of that compound's anticancer activity. The $CHCl_3$ layer was purified via repeated column chromatography and recrystallization. The two compounds isolated from $CHCl_3$ layer were identified via NMR spectroscopic analysis as (10E) 1,10-heptadecadiene-4,6-diyne-3,8,9-triol (Comp. I) and anomalin (Comp. II). (10E) 1,10-heptadecadiene-4,6-diyne-3,8,9-triol was the first report from the roots of P. japonicum. MTT assays were conducted to evaluate the in vitro cytotoxic activities of Compounds I and II against the following human cancer cell lines: HeLa, HepG2, SNU-16, and AGS. Comp. I evidenced the most profound cytotoxic activity against HepG2 cells ($IC_{50}=6.04{\mu}g/mL$), and Comp. II exhibited the most profound cytotoxic activity against SNU-16 cells ($IC_{50}=18.24{\mu}g/mL$) among the human cancer cell lines tested in this study. However, no significant cell death was observed in the CCD-25Lu human normal lung fibroblast cells. Quantitative analysis using UPLC (Ultra Performance Liquid Chromatography) showed that the roots of P. japonicum contained 0.015 (Comp. I) and 1.69 mg/g (Comp. II) of these compounds.

• Biomolecules & Therapeutics
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• 제23권4호
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• pp.379-385
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• 2015

The eye irritation potential of drug candidates or pharmaceutical ingredients should be evaluated if there is a possibility of ocular exposure. Traditionally, the ocular irritation has been evaluated by the rabbit Draize test. However, rabbit eyes are more sensitive to irritants than human eyes, therefore substantial level of false positives are unavoidable. To resolve this species difference, several three-dimensional human corneal epithelial (HCE) models have been developed as alternative eye irritation test methods. Recently, we introduced a new HCE model, MCTT HCE$^{TM}$ which is reconstructed with non-transformed human corneal cells from limbal tissues. Here, we examined if MCTT HCE$^{TM}$ can be employed to evaluate eye irritation potential of solid substances. Through optimization of washing method and exposure time, treatment time was established as 10 min and washing procedure was set up as 4 times of washing with 10 mL of PBS and shaking in 30 mL of PBS in a beaker. With the established eye irritation test protocol, 11 solid substances (5 non-irritants, 6 irritants) were evaluated which demonstrated an excellent predictive capacity (100% accuracy, 100% specificity and 100% sensitivity). We also compared the performance of our test method with rabbit Draize test results and in vitro cytotoxicity test with 2D human corneal epithelial cell lines.

• 생약학회지
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• 제36권3호통권142호
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• pp.201-204
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• 2005

The rhizome extract of Atractylodes japonica Koidzumi(Compositae) exhibited a particular inhibition on the proliferation of cultured human tumor cell lines, in vitro. Thus, the intensive phytichemical investigation of the MeOH extract of Atractylodes japonica have been conducted by the way of activity-guided purification. The repeated column chromatographic separation of the n-hexane soluble part of extract resulted in the isolation of four sesquiterpenes (1-4) and a polyacetylene component (5). Chemical structures of them were identified as atractylon (1), atractylenolide Ⅰ(2), atractylenolide Ⅲ(3), eudesma-4(15),7(11)-dien-8-one (4) and 1,3-diacetyl-atractylodiol (5) by spectroscopic means. Among the isolates, compound 2-4 were shown to give moderate inhibitory effect in a dose dependent manner on the proliferation of cultured human tumor cell lines such as A549 (non small cell lung), SK-OV-3 (ovary), SK-MEL-2 (melanoma), XF498 (central nerve system) and HCT 15(colon), respectively.

• 한국축산식품학회지
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• 제41권4호
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• pp.687-700
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• 2021

This study performed to evaluate the applicability of functional dairy food materials by comparing the calcium solubilization ability and anti-inflammatory effects of hydrolyzed casein protein. Commercial enzyme (Alcalase^®; Neutrase^®; Protamex^®; Flavourzyme^®) was added to the 10% casein solution to prepare the casein hydrolysates. Samples obtained every hour [1:200 (w/v)]. According to results of measuring the degree of hydrolysis (DH), all of four enzymatic hydrolysates increased rapidly from 30 to 40 min, and after 150 min, there were no change. Protamex^® and Neutrase^® had the highest DH compared to others enzymatic hydrolysates. After that, peptides obtained throughout a preparative liquid chromatography system. In the calcium solubility experiments, neutrase fraction (NF) 4 and NF7 showed similar activities with casein phosphopeptide (CPP). In vitro cell experiments showed that no cytotoxicity except for NF6. Also, the production of nitric oxide (NO) inhibited as the concentration of fraction samples increased. The cytokine (IL-1α, IL-6, and TNF-α) production was lower than lipopolysaccharide (+) group significantly. Therefore, the possibility of anti-inflammatory activity found in the hydrolyzed samples. According to the above experiments, NF3 and Protamex Fraction (PF) 3 selected. Amino acids selected throughout an AccQ-Tag system. As a result, 17 species of amino acids and several species of unknown amino acids identified. Both fractions had the highest content of phenylalanine. This study identified the potential of biologically active and functional peptides derived from casein that affect the food and dairy industry.

• 한국미생물·생명공학회지
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• 제50권1호
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• pp.22-30
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• 2022

Haliotis discus hannai called abalone, is the valuable marine mollusks and the by-products of abalone processing are viscera. Brownish abalone male viscera (AMV), which have not been reported as having anti-inflammatory effects, was extracted with acetone and fractionated by different six acetone/hexane ratios (0, 10, 20, 30, 40, and 100%) using a silica column via in vitro ABTS and DPPH radical and nitric oxide (NO) production assay-guided fractionation. Among the fractions, the acetone/hexane ratio 40%, A40 exhibited the most potent radical scavenging activities and inhibition of lipopolysaccharide (LPS)-induced NO production without cytotoxicity. A40 inhibited LPS-induced intracellular reactive oxygen species (ROS) production in a dose-dependent manner. Western blot analysis revealed that A40 down-regulated the activation of NF-κB, MAPK (ERK 1/2, p-38, and JNK), and inflammatory enzymes, inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2. Moreover, this fraction inhibited the generation of pro-inflammatory cytokines such as interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α. These results suggested that AMV containing A40 with anti-inflammatory and anti-oxidantive effects, is the effective therapeutic and functional material for treating inflammatory disorders.

• 생명과학회지
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• 제34권5호
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• pp.339-355
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• 2024

폐는 생리학적 기능과 해부 조직학적 구조 측면을 통합적으로 고려하여 분석해야만 하는 매우 복잡한 조직이기 때문에 폐질환의 병리학적 연구와 흡입독성 평가에 현재까지 주로 동물모델을 사용하고 있다. 그러나 실험동물 윤리와 동물복지를 이유로 점차적으로 실험동물 수를 줄이자는 전세계적인 움직임에 맞춰 생체 외 동물실험 대체법들이 집중적으로 개발되고 있다. 특히 경제협력개발기구(OECD)와 미국 환경보호청(USEPA)은 2030년대 이후, 동물실험을 금지하기로 잠정적으로 합의함에 따라 의생명공학과 제약 분야에서 생체 외 흡입 독성 및 폐질환 모델들을 확립하고 개발된 모델을 이용한 평가 법들의 표준화 연구가 활발하다. 그 모델 중에 예를 들어, 생체칩(organ-on-a-chip, OoC) 및 오가노이드(organoid) 모델은 3차원 바이오 프린터, 미세 유체 시스템, 인공지능(artificial intelligent) 기술들과 접목되어 연구되고 있다. 이러한 생체 장기를 모방한 복합 장기 생체 외 모델링 시스템은 개체 차이를 가지는 생체 내 동물 실험에 비해 복잡한 생물학적 환경을 보다 정확하게 모방할 수 있을 것으로 기대되고 있으나 생체 모방성, 재현성, 민감성, 기반 데이터베이스의 부족 등 아직은 여러 한계점도 가지고 있다. 따라서 본 리뷰 논문에서는 만능성 줄기 세포 또는 암세포를 이용한 폐포, 폐 공기액 인터페이스(air-liquid interface, ALI) 시스템, 트랜스웰 멤브레인(transwell membrane)을 포함하여 폐 OoC 및 오가노이드의 최근 생체 외 폐 시스템 연구결과들과 AI와 접목된 인실리코(in silico) 폐 모델링에 대한 결과들의 현황을 살펴보고자 한다.

• 한국식품조리과학회지
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• 제15권3호
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• pp.231-237
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• 1999

닥나무 추출물의 항암효과를 검토하기 위하여 닥나무뿌리 껍질 methanol 추출물로부터 얻은 분획물(hexane, chloroform, ethylacetate, butanol, aqueous 층)들의 0.05∼1.0mg/ml 농도에 대해서 MTT assay를 통한 세포독성 측정 및 암세포 증식억제 효과를 in vitro로 검색하였고, in vivo에서 BALB/c mouse를 이용한 고형암 성장저지 실험 및 비장 장기 중량변화의 항암실험을 행하여 다음과 같은 결과를 얻었다. 세포독성 실험에 의한 MTT assay 결과 모든 분획물들은 농도 의존성으로 효과가 나타났으며 1.0 mg/ml 농도에서 ethylacetate층은 sarcoma 180 cell에 대하여 75.7%의 가장 높은 세포독성 효과를 나타내었고 그 다음 aqueous, hexane, butanol, chloroform 층 순으로 각각 62.7%, 58.7%, 52.6%, 40.1%의 효과를 보였다. 각 분획물들에 의한 sarcoma 180 cell에 증식억제 효과는 배양일(1, 2, 3, 6일)이 증가함에 따라 암세포수는 증가하였으며 처음 1일째에 암세포 증식억제가 가장 많이 되었다. 1.0mg/ml 농도로 시료를 첨가하여 6일간 배양했을 때 ethylacetate 층은 60%의 억제율로 가장 현저 한 활성을 보였고 aqueous(42%), hexane(31%), butanol(30%), chloroform(19%)층 순으로 암세포 증식억제율을 나타내었다. 또한 암세포 증식에 대한 현미경 관찰을 해 본 결과 ethylacetate층은 대조군에 비하여 암세포 증식억제 정도와 형태 변화가 두드러지게 나타났다. BALB/c mouse에서 sarcoma 180 cell에 대한 각 분획물들의 고형암 성장저지효과는 1.0mg/kg으로 시료투여했을 때 ethylacetate 층은 대조군에 비하여 41%의 유의적인 저지율을 나타냈으며(p<0.05), 그 다음은 유의적이지는 않으나 aqueous 층 처리구가 39%,의 저지율을 나타내었다. 비장의 체중에 대한 중량변화는 대조군의 경우 1.08%인데 비해, ethylacetate층 처리군은 2.10%로 증가하였고 나머지 처리군에서는 대조군과 차이를 나타내지 않았다(p<0.05). 한편, mouse의 체중 변화는 대조군과큰 차이가 없었으므로 시료 추출물로 인한 체중 및 장기의 독성 문제는 없을 것으로 사료된다. 이상의 연구 결과로 닥나무 뿌리 껍질 methanol 추출물로부터 얻은 분획물중 ethylacetate와 aqueous 층은 세포독성 효과와 암세포 성장억제 효과 및 고형암 성장 저지 효과가 크게나타났으며, 체중에 대한 면역관련 장기인 비장의 중량증가로부터 면역계 활성과도 관계가 있을 것으로 사료된다. 따라서 닥나무 뿌리 껍질의 효능이 큰 분획물로부터 항암활성 물질을 분리 정제하고 기전확립의 연구가 앞으로 더욱 보충되어야 할 것이며, 암예방을 위한 기능성 식품 및 치료제의 개발에 대한 가능성 확인을 본 연구를 통하여 확인할 수 있었다.

• 생명과학회지
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• 제16권6호
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• pp.904-910
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• 2006

수지상세포는 종양면역에서 필수적인 강력한 CTL 반응을 개시할 수 있는 유일한 세포이다 . 특히 외인성 종양항원에 대한 CTL 반응 유도는 활성화된 수지상세포의 IL-12 분비를 통한 CD4+ helper T세포의 cross-priming을 필요로 한다. 그러나 최근에 활성화된 수지상세포는 $Th_1$ 면역반응을 유도하지만 활성화 시간이 경과함에 따라 오히려 $Th_2$반응을 유도 할 수 있다. 따라서 본 연구에서는 OVA를 종양항원 모델로 설정하여 종양특이적인 CTL 반응을 형성하기 위한 최적의 수지상세포 활성화 조건을 조사하였다. 마우스 골수세포에 서 수지상세포로의 분화는 항원제시 기능을 위한 표면분자의 발현 측면에서 볼 때 배양 6일-7일 정도가 적합하였다. 수지상세포의 IL-12 생성능은 배양 6일 이상, OVA 항원 탑재 8시간 이상의 경우에 연이은 LPS 성숙자극으로 오히려 감소하는 경향을 보였다. 즉 배양 6일의 수지상세포에 OVA 항원 탑재를 8시간 수행한 경우(8-h DC)가 in vitro에서의 IL-12생성능, ex vivo에서의 세포내 $IFN-{\gamma}$를 발현하는 CD8+ T세포의 증가 및 OVA 특이적인 세포독성효과 등에서 가장 좋은 결과를 보였다. 또한 in vivo에서 종양 치료 및 예방효과에서도 8-h DC로 면역한 경우에 가장 우수한 종양형성 억제 효과와 생존기간 연장효과를 보였다. 현재 대부분의 수지상 세포를 이용한 항종양 백신에서 항원 탑재반응을 24시간 동안 수행하고 있으나, 본 실험의 결과로 볼 때, 8시간의 in vitro 항원 탑재가 보다 효과적인 종양특이적 CTL 반응과 항종양 면역반응을 유도함을 알 수 있다. 결론적으로 본 연구를 통하여 8시간 이상의 항원접촉은 수지상세포의 기능적 활성능력을 오히려 고갈시킬 수 있음을 제시한다.