• 한국식품과학회지
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• 제32권4호
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• pp.964-969
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• 2000

랩 등 PVC 제품의 가소제인 DEHA의 에스트로겐 활성을 검색하기 위하여 내분비계 장애작용 중 에스트로겐성 작용을 검색하는 대표적인 방법인 in vitro E-screen assay와 OECD 및 미국 EPA EDSTAC에서 권고하고 있는 in vivo 난소절제 랫드를 이용한 자궁 비대반응시험(uterotrophic assay)을 실시하였다. 시험결과 $DEHA(5{\times}10^{-9}{\sim}5{\times}10^{-4}\;M)$는 MCF-7세포의 증식을 유발하지 않았고 난소절제 랫드에 200 mg/kg/day 용량까지 투여하여도 자궁 및 질 무게에 영향을 미치지 않았다. 따라서 본 실험결과 DEHA는 E-screen assay 및 자궁비대반응시험에서 에스트로겐성 작용이 없는 것으로 확인되었으나, 이들 물질의 에스트로겐성에 대한 결론을 내리기 위해서는 여러 종류의 in vitro 시험 data 및 in vivo 시험 data가 더 보충되어야 할 것으로 생각된다.

• 한국식품영양과학회지
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• 제28권1호
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• pp.240-245
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• 1999

Growth inhibitory effect of doenjang(Korean soypaste) methanol extracts in SRB assay using AGS human gastric adenocarcinoma cell, Hep 3B human hepatocellular carcinoma cell and HT 29 human colon cancer cell was studied. The treatment of doenjang methanol extracts(2mg/assay) to the AGS, Hep 3B and HT 29 cancer cells inhibited the growth of the cancer cells by 55%, 60%, and 71%, respectively. Doenjang methanol extracts exhibited the highest inhibitory effect among other soybean fermented foods and original materials in the SRB assay. In addition, to separate active compounds of doenjang methanol extracts, we fractionated the doenjang with hexane, methanol, dichloromethane, ethylacetate and butanol. Growth inhibitory effect on the AGS, Hep 3B, HT 29 and MG 63 cancer cells was the highest in the fractions of dichloromethane and ethylacetate among other solvent fractions of the doenjang. These results showed that some compounds contained in the fractions of dichloromethane and ethylacetate might play a role on the anticanceric effect of doenjang.

• Environmental Analysis Health and Toxicology
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• 제15권1_2호
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• pp.13-18
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• 2000

A few in vitro methods were developed to compare the result on the phototoxicity of phenothiazines. By the MTT assay, the Candida test, and the RBC photohemolysis, the phototoxicities of UVA and UVB irradiation were measured. This paper presents the comparisons of methods which are effective to measure the phototoxicities of the chemicals causing phototoxicity and photoallergy. The tested chemicals of phenothiazines include Chlorpromazine, Promethazine, Perphenazine, Chlorprothixene, Trifluoperazine and Thioridazine. Each chemical represented variable results according to the test methods. MTT assay shows the most sensitive method.

• Journal of Nutrition and Health
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• 제49권5호
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• pp.277-287
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• 2016

본 연구는 제5기 2차년도 국민건강영양조사 결과를 활용하여 한식 식품군의 총 페놀 함량, in vitro 항산화활성 및 인체세포를 이용한 ex vivo DNA 손상 감소효과를 비교하고, 각 지표간의 상관성을 분석하며, 한식의 총 식사 항산화능에 대한 각 식품군의 기여도를 알아보기 위해 수행되었다. 제5기 2차년도 국민건강영양조사 결과를 바탕으로 한식의 식물성 식품을 10가지 식품군 (곡류, 과일류, 채소류, 견과류, 김치류, 해조류, 감자류, 버섯류, 두류, 오일류)으로 분류한 후 각 식품군별로 총 섭취량의 1% 이상 섭취한 식품 84종을 한식의 식물성 식품으로 최종 선정하였다. 각 식품군의 총 페놀함량을 측정하였고, DPPH radical scavenging assay, TEAC assay, $ORAC_{ROO{\cdot}}$ assay를 사용하여 in vitro 항산화능을 측정하였다. 한식의 식품군별 항산화능 (dietary antioxidant capacity, DAC)은 in vitro 항산화활성 평균값과 각 식품군의 1일 섭취량을 고려하여 계산하였고 한식 TDAC는 각 식품군의 DAC로의 합으로 구하였으며, TDAC에 대한 각 식품군 항산화능의 기여도를 평가하였다. 인체 임파구에서의 ex vivo DNA 손상 정도는 comet assay를 사용하여 평가하였다. 한식 식품군의 총 페놀함량은 버섯류, 과일류, 채소류, 해조류, 김치류 등의 순으로 높았으며, 3가지 in vitro 실험법을 평균한 식품군의 항산화활성 순위는 버섯류, 해조류, 채소류, 김치류, 과일류 등의 순이었다. 각 식품군의 항산화활성에 식품섭취량을 고려하여 계산한 한식의 TDAC에 대한 식품군의 항산화능 기여도는 곡류가 33.4%로 가장 높았으며, 과일류 (23.9%), 채소류 (12.7%), 김치류 (11.2%) 등의 순으로 나타났다. 인체 임파구에서 ex vivo DNA 손상 보호효과는 버섯류에서 가장 높았으며, 그 다음 채소류, 과일류, 해조류, 김치류의 순으로 나타났다. 각 식품군의 페놀함량과 in vitro 항산화 활성, 그리고 ex vivo DNA 보호효과의 순위가 비슷하게 나타났으며 각 지표간의 상관성은 매우 높았다. 한식 식품군 중 버섯류, 과일류, 채소류, 해조류에서 총 페놀함량과 항산화 활성, DNA 손상 보호효과가 높게 나타났다. 각 식품군의 총 페놀함량과 in vitro 항산화 활성, ex vivo DNA 보호효과 지표 간의 상관성은 매우 높았다. 한식의 TDAC에 대한 식품군별 항산화능 기여도는 곡류가 가장 높았고, 그 다음이 과일류, 채소류, 김치류의 순이었다. 이러한 결과는 앞으로 한식의 우수성을 항산화 측면에서 밝히는데 매우 중요한 기초자료로 활용될 수 있을 것이다.

• 한국식품영양과학회지
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• 제33권9호
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• pp.1486-1491
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• 2004

식품오염 관련 중금속들의 에스트로겐성을 in vitro 와 in vivo 분석방법을 병행하여 평가하였다. 분석방법은 1) estrogen receptor dependent transcriptional expression 분석법, 2) E-screen assay 그리고, 3) 마우스 자궁비대시험 (uterotropic assay)을 사용하였다. 시험에 사용한 물질로는 $17\beta$-estradiol, diethylstilbestrol(DES), arsenic oxide, bis (tri-n-butyltin), cadmium chloride, chromium chloride, lead acetate, mercuric chloride을 사용하였다. Estrogen receptor dependent transcriptional expression 분석 결과, bis(tri-nbutyltin) > cadmium chloride > chromium chloride 순으로 에스트로겐성이 크게 나타났으며, mercuric chloride, lead acetate, arsenic oxide는 거의 나타나지 않았다. E-screen test 결과, bis(tri-n-butyltin) > cadmium chloride > chromium chloride 순으로 에스트로겐성이 크게 나타났으며, mercuric chloride, lead acetate, arsenic oxide는 거의 나타나지 않았다. 자궁비대시험 결과도 마찬가지로 bis(tri-nbutyltin), cadmium chloride, chromium chloride은 자궁중량 비대를 크게 초래하였으며, 반면에 mercuric chloride, lead acetate, arsenic oxide는 그러한 효과가 미약하거나 없었다. 세 분석방법 결과 bis(tri-n-butyltin), cadmium chloride, chromium chloride 순으로 에스트로겐성이 크게 나타났다. 이러한 결과는 최근 bis(tri-n-butyltin)과 cadmium chloride이 에스트로겐성이 있다는 다른 연구결과들과 잘 일치하며, 또한 크롬화합물도 에스트로겐성이 있다는 것을 새롭게 제시하고 있다. 본 연구는 세 단계 수준(전사활성화단계, 세포증식작용, in vivo assay)의 분석을 병행함으로써 수많은 중금속의 에스트로겐성을 효과적으로 평가할 수 있다는 것을 제시해주고 있다.

• 한국양식학회지
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• 제16권3호
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• pp.196-201
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• 2003

Sessile organisms such as the oyster Crassostrea gigas have been given much attention as a potential biomonitoring indicator to assess the impact of toxicants on aquatic organism. In this study, we exposed cells isolated from gill of oyster (Crassostrea gigas) to hydrogen peroxide in vitro. In addition oysters were in vivo exposed to pyrene and benzo(a)pyrene at various concentrations for 2 weeks. Comet assay was used to detect DNA single strand breaks and to investigate the application of this technique as a tool for aquatic biomonitoring. Hydrogen peroxide increased DNA single strand break with increasing concentration after 30 minutes exposure in vitro. Pyrene and benzo(a)pyrene caused DNA damage only at very high concentration (100 $\mu\textrm{g}$/L or 1000 $\mu\textrm{g}$/L) at two week exposure in vivo. DNA damage was relatively higher at hemocyte than at gill. It suggested that metabolized PAHs are transferred to hemolymph from digestive gland which have a relatively high enzyme activity, and attacked the DNA of hemocyte, while gill accumulated PAHs without degrading them to their metabolites due to low enzyme activity at gill. Both in vitro and in vivo exposure experiments showed that the comet assay is an effective tool on screening whether the organism are exposed to genotoxic contaminants.

• Asian-Australasian Journal of Animal Sciences
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• 제29권11호
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• pp.1608-1615
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• 2016

This study was conducted to determine the solubility of copper (Cu) in two sources of copper(II) sulfate ($CuSO_4$) including monohydrate and pentahydrate and three sources of dicopper chloride trihydroxide (dCCTH) including ${\alpha}$-form (dCCTH-${\alpha}$), ${\beta}$-form (dCCTH-${\beta}$), and a mixture of ${\alpha}$- and ${\beta}$-form (dCCTH-${\alpha}{\beta}$) at different pH and a 3-step in vitro digestion assay for pigs. In Exp. 1, Cu sources were incubated in water-based buffers at pH 2.0, 3.0, 4.8, and 6.8 for 4 h using a shaking incubator at $39^{\circ}C$. The $CuSO_4$ sources were completely dissolved within 15 min except at pH 6.8. The solubility of Cu in dCCTH-${\alpha}$ was greater (p<0.05) than dCCTH-${\beta}$ but was not different from dCCTH-${\alpha}{\beta}$ during 3-h incubation at pH 2.0 and during 2-h incubation at pH 3.0. At pH 4.8, there were no significant differences in solubility of Cu in dCCTH sources. Copper in dCCTH sources were non-soluble at pH 6.8. In Exp. 2, the solubility of Cu was determined during the 3-step in vitro digestion assay for pigs. All sources of Cu were completely dissolved in step 1 which simulated digestion in the stomach. In Exp. 3, the solubility of Cu in experimental diets including a control diet and diets containing 250 mg/kg of additional Cu from five Cu sources was determined during the in vitro digestion assay. The solubility of Cu in diets containing additional Cu sources were greater (p<0.05) than the control diet in step 1. In conclusion, the solubility of Cu was influenced by pH of digesta but was not different among sources based on the in vitro digestion assay.

• Toxicological Research
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• 제14권3호
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• pp.315-320
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• 1998

In vitro skin iritation of anti-inflammatory drugs was investigated in terms of the cytotoxicity method to human skin fibroblast cells. Five anti-inflammatory drugs (Diclofenac, Naproxen, Meclofenamic acid, Ibuprofen and Fnoprofen) which are commercially available as oral preparations or injections were tested. The cytotoxicity of 5 chemicals was evaluated by using MTT[tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay. NRU (neutral red uptake) assay and Alamar Blue assay after fibroblast cells had been exposed to the chemicals for 24 hours or 489 hours. The $IC_{50}$ values of the chemicals showed the comparative strength of cytotoxicity as following order of Meclofenamic acid>Diclofenac>Fenoprofen>Ibuprofen>Naproxen. The values of $IC_{50}$ determined by Alamar Blue assay were lower than those of MTT and NRU assay. These data suggest Alamar Blue assay can be useful method for assessing in vitro skin irritation potential of anti-inflammatory drugs.

• Toxicological Research
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• 제16권2호
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• pp.141-146
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• 2000

Phthalate esters are used extensively as a plasticizer in the manufacture of plastic products such as PVC bags and medical devices. Recently, phthalate esters have been shown to induce endocrine system mediated responses. However. only a Jew studies have been conducted for estrogenic activity of phthalate esters. In this study estrogenic activities of seven phthalate esters. butyl benzyl phthalate (BBP), di(2-ethylhexyl) phthalate (DEHP), di-n-butylphthalate (DBP), diethylphthalate (DEP), di-n-pentylphthalate (DPP), di-n-propylphthalate (DPrP) and dicyclohexylphthalate (DCHP), were examined in vitro using E-screen assay and competitive binding assay. From the E-screen assay, BBP. DEHP. DBP and DEP showed weak estrogenic activity at the concentration of 5 $\mu\textrm{M}$. The relative proliferative effect (RPE) and the relative proliferative potency (RPP) were 50~70% and 0.01%. respectively, when compared with 500 pM of 17$\beta$-estradiol (E2). In competitive binding assay with the rat uterine estrogen receptor (ER), BBP and DEP showed weak binding potency ［(l/$10^4$~1/$10^5$ of E2］ while DEHP and DBP scarcely bound to ER. These results suggest that some phthalate esters have weak estrogenic activities in vitro.

• Environmental Analysis Health and Toxicology
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• 제17권3호
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• pp.253-259
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• 2002

The phototoxicity inhibitory activity of 15 natural products having antiinflammatory effect was screened by three in vitro methods: yeast growth inhibition test with Candida albicans, RBC photohemolysis and MTT assay. We induced phototoxic reaction by irradiating UVA (365 nm) on chlorpromazine (CPZ) that has been widely documented as phototoxic agent in clinical and experimental studies and then observed the effects of the natural products after treating them with CPZ. In yeast growth inhibition test, X. stramonium showed the inhibitory effect on the UVA phototoxicity and E. officinalis, Yeast, P. suffruticosa showed phototoxicity inhibitory effect in that their ％ hemolysis compared with control were 36.14${\pm}$ 2.69, 42.82${\pm}$1.35, 36.41${\pm}$0.48 on UVA. In MTT assay, all tested natural products increased cell viability compared with the control.