• 제목/요약/키워드: in vitro detection system

검색결과 71건 처리시간 0.024초

In vitro 검출시스템을 이용한 한약재 추출물로부터의 에스트로겐 활성의 검증 (Verification of Estrogenic Activities in Ethanol Extracts of Oriental Herbal Medicines using In vitro Detection System)

  • 이상현
    • 동의생리병리학회지
    • /
    • 제17권4호
    • /
    • pp.1054-1058
    • /
    • 2003
  • In order to evaluate the direct effect of estrogenic compounds in oriental herbal medicines, the estrogenic activity was measured using an in vitro detection system. For this system, human breast cancer cell line MCF7 was transfected using an estrogen responsive CAT reporter plasmid. Estrogenic activities of Platycodi radix, Astragali radix and Glycyrrhizae radix were evaluated using this system. Estrogenic activity of a 500 ㎍/ml ethanol extract of Platycodi radix was as same as that of a 10/sup -8/ M standard solution (17β-estradiol) and activity of a 50 ㎍/ml ethanol extract was between those of a 10/sup -8/ M and a 10/sup -9/ M standard solutions. In addition, estrogenic activity of a 50 ㎍/ml ethanol extract of Platycodi radix was as same as that of a 10/sup -10/ M standard solution. The same activity patterns were observed in the system which was treated by Astragali radix or Glycyrrhizae radix extracts. The most effective activity was observed in a system which was treated by Platycodi radix extract, but the least activity was observed by Glycyrrhizae radix extract. In this result, it was confirmed that Platycodi radix, Astragali radix and Glycyrrhizae radix extracts possess estrogenic compounds.

테스토스테론 물질 검출을 위한 in vitro TCD 시스템 구축 (Establishment of an In Vitro TCD (Testosterone Compound Detection) System)

  • 이동근;조정권;이상현
    • 생명과학회지
    • /
    • 제29권10호
    • /
    • pp.1159-1163
    • /
    • 2019
  • 남성호르몬 감소와 관련된 남성갱년기에 대한 관심이 고조되고 있지만, 남성호르몬의 정량을 위해 항체를 이용하는 고가의 kit가 이용되고 있다. 본 연구에서는 in vitro 전사 활성 시험법을 이용하여 남성 스테로이드호르몬의 활성 혹은 농도를 검증하는 시스템을 구축하였다. 테스토스테론-AR (androgen receptor) 복합체와 반응하는 ARE-AdE1bTATA 염기서열이 삽입되고 리포터로 luciferase를 발현하는 테스토스테론 유사활성 검증 리포터 플라스미드인 pGL2-Neo-ARE-AdE1BTATA를 제조하고, 인체 전립선암 세포인 LNcap-LN3 세포에 stable transfection을 실시하였다. 구축된 LNcap-LN3/pGL2-Neo-ARE-AdE1BTATA TCD (testosterone compound detection) 시스템은 표준물질인 테스토스테론의 $10^{-13}{\sim}10^{-8}M$ 범위에서 농도 증가에 비례하는 정량성을 보였다. 이 연구에서 확립된 in vitro TCD 시스템을 이용하면 천연물 유래 테스토스테론 유사물질 및 테스토스테론 저하물질의 대량 탐색 등이 가능할 것이므로, 건강기능성 식품이나 의약품 신소재의 개발에 기여할 것이다.

In vitro 검출시스템을 이용한 해양생물 추출물로부터 에스트로겐 활성 검증 (Verification of Estrogenic Activity in Ethanol Extracts of Marine Organisms Using in vitro Test System.)

  • 하종명;이상현
    • 생명과학회지
    • /
    • 제13권6호
    • /
    • pp.799-804
    • /
    • 2003
  • 천연물에 포함되어 있는 에스트로겐 효과를 가지는 성분의 생체 내에서의 직접적인 효과를 검정하기 위하여, 에스트로겐 수용체를 발현하는 것으로 알려진 인체 유방암 세포주 MCF7에 에스트로겐에 반응성을 나타내도록 고안된 CAT 리포터 플라스미드를 도입한 in vitro 검출시스템을 사용하여 에스트로겐 활성을 측정하였다. 이미 여러 분야에서 그 기능성의 연구가 활발히 진행되고 있는 광합성 조류인 스피루리나(spirulina)와 파래 등의 해양식물을 대상으로 에스트로겐 반응 리포터 시스템을 이용하여 에스트로겐 활성을 측정하였다. 그 결과, 스피루리나 에탄올 추출물의 CAT 활성은 $500\mug/ml\; 와\; 50 \mug/ml$의 농도에서 표준물질인 $17\beta-estradiol$의 농도 $10^{-8}$ M과 비슷한 정도의 에스트로겐 활성 효과를 나타내었고, $5\mug/ml$의 농도에서는 표준물질인 $17\beta-estradiol$의 농도 $10^{-10}$ M과 비슷한 정도의 에스트로겐 활성 효과를 나타내었다. 하지만 파래 추출물의 경우에는 에탄올을 처리한 대조군과 비교하여 유의한 CAT활성 변화를 나타내지 않았다 한편, 불가사리와 새우 등의 해양동물을 대상으로 한 실험에서는 에탄올을 처리한 대조군과 비교하여 유의한 CAT 활성 변화를 나타내지 않았다. 이 연구 결과로 광합성 조류인 스피루리나에 에스트로겐 활성을 효과적으로 나타내는 생리활성성분이 포함되어 있을 수 있다는 가능성을 확인하였다.

iNOS 발현 검출을 위한 in vitro 시스템의 확립 및 적송잎 추출물에 의한 저해효과 검증 (Establishment of In vitro Detection System for iNOS Expression and the Verification of Suppressive Effect by Pine Needle Extract)

  • 김남영;장혜지;이동근;장민경;이승우;전명제;김미향;김성구;이상현
    • KSBB Journal
    • /
    • 제26권2호
    • /
    • pp.172-176
    • /
    • 2011
  • This study was aimed to verify suppressive effect of pine-needle extract on lipopolysaccharide (LPS)-induced inducible nitric oxide synthase (iNOS) expression. In order to evaluate suppressive effect on iNOS expression, RAW 264.7 cells were stably transfected using an iNOS promoterluciferase reporter plasmid yielding RAW 264.7/pGL2-NeomiNOS_ pro11 cells. Established in vitro detection system revealed to diminish LPS-induced iNOS expression by 0.1~500 ${\mu}g/mL$ of saponin at the concentration-dependant manner. Pine needle extract also diminished LPS-induced iNOS expression to 92 and 88% at 500 and 50 ${\mu}g/mL$, respectively. These results suggest that the in vitro detection system developed here could be useful for the verification of suppressive materials on iNOS expression and pine needle extract could be used for the development of functional foods.

The Isolation of Acetylcholinesterase Inhibitory Constituents from Lycoris radiata using On-line HPLC-biochemical Detection System

  • Yang, Hee-Jung;Yoon, Kee-Dong;Chin, Young-Won;Kim, Young-Choong;Kim, Jin-Woong
    • Natural Product Sciences
    • /
    • 제16권4호
    • /
    • pp.228-232
    • /
    • 2010
  • Bioactivity-guided fractionation using on-line HPLC biochemical detection system on $CHCl_3$-soluble fraction of Lycoris radiata led to the isolation of deoxylycorenine (1), O-demethylhomolycorine (2), galanthamine (3), lycoramine (4), mixture of $6{\alpha}$-and $6{\beta}$-haemanthidine (5), and lycorine (6), identified by spectroscopic data and physicochemical property. Among the isolated compounds, 1, 3 and 6 showed acetylcholinesterase inhibitiory activities with $IC_{50}$ values of 18.0, 12.0 and $16.6\;{\mu}M$, respectively, in in vitro colorimetric microplate assay.

Evaluation of the Genetic Toxicity of Synthetic Chemicals (XVI) - in vitro Mouse Lymphoma Assay with 3 chemicals -

  • Kim, Youn-Jung;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
    • /
    • 제2권4호
    • /
    • pp.244-250
    • /
    • 2006
  • The detection of many synthetic chemicals used in industry that may pose a genetic hazard in our environment is of great concern at present. Since these substances are not limited to the original products, and enter the environment, they have become widespread environmental pollutants, thus leading to a variety of chemicals that possibly threaten the public health. In this respect, to regulate and to evaluate the chemical hazard will be important to environment and human health. The genotoxicity of 3 synthetic chemicals was evaluated in L5178Y $tk^{+/-}$ mouse lymphoma cells in vitro. 9H-carbazole (CAS No. 86-74-8) did not induce significant mutation frequencies both in the presence and absence of metabolic activation system. 1, 3-Dichloro-2-propanol (CAS No. 96-23-1) revealed a significant increase of mutation frequencies in the range of $625-373\;{\mu}g/mL$ in the absence of metabolic activation system and $157-79\;{\mu}g/mL$ in the presence of metabolic activation system. And also, fenpropathrin (CAS No. 64257-84-7) appeared the positive results only in the absence of metabolic activation system. Through the results of MLA tk assay with 3 synthetic chemicals in L5178Y cells in vitro, we may provide the important clues on the genotoxic potentials of these 3 chemicals.

가상내시경의 개발 및 가상 대장내시경으로 적용 시 성능평가 (Development of Virtual Endoscopy and Evaluation of Performance as a 3D Virtual Colonoscopy)

  • 김정훈;이상훈;고성호;김상준
    • 대한의용생체공학회:의공학회지
    • /
    • 제24권2호
    • /
    • pp.69-75
    • /
    • 2003
  • Virtual colonoscopy is one of the Powerful tool for non-invasive colon examination and many in-vitro and in-vivo studies have shown its accuracy in Polyp or adenoma detection. But most of virtual colonoscopy requires high quality workstation and software and its cost is high to setup whole system. We developed PC-based 3D model creation and navigation program which has diverse functions. It can be easily installed to PC and connected to network system. The performance. when used as a virtual colonoscopy. is evaluated by calculating sensitivity of detection for the simulated polyp which is artificially made inside the Pig's colon and checked its clinical feasibility, Its total sensitivity is 76%. Grouping according to Polyps diameter, the sensitivity for detection of polyps 10 ㎜ or larger was 100%(40 of 40); 5.0-9.9 ㎜, 90.0(90 of 100): and smaller then 5 ㎜. 36.7%(22 of 60).

Evaluation of the Genetic Toxicity of Synthetic Chemicals (IV) - in vitro Chromosomal Aberration Assay with 18 Chemicals in Chinese Hamster Lung Cells -

  • Ryu, Jae-Chun;Kim, Kyung-Ran;Kim, Youn-Jung
    • 한국환경성돌연변이발암원학회지
    • /
    • 제22권3호
    • /
    • pp.149-156
    • /
    • 2002
  • The detection of many synthetic chemicals used in industry that may pose a genetic hazard in our environment is of great concern at present. Since these substances are not limited to the original products, and enter the environment, they have become widespread environmental pollutants, thus leading to a variety of chemicals that possibly threaten the public health. In this respect, to regulate and to evaluate the chemical hazard will be important to environment and human health. The clastogenicity of 18 synthetic chemicals was evaluated in Chinese hamster lung fibroblast cells in vitro. 4-Chloro-3,5-dimethyl phenol (CAS No. 88-04-0) induced chromosomal aberrations with significance at the concentration of 15.7 $\mu\textrm{g}$/$m\ell$ both in the presence and absence of metabolic activation system. Phenoxybenzene (CAS No. 101-84-8) which is one of the most cytotoxic chemical among 18 chemicals tested revealed no clastogenicity in the range of 0.11-0.43 $\mu\textrm{g}$/$m\ell$ both in the presence and absence of metabolic activation system. From the results of chromosomal aberration assay with 18 synthetic chemicals in Chinese hamster lung cells in vitro, 4-chloro-3,5-dimethyl phenol (CAS No. 88-04-0) revealed weak positive clastogenic results in this study.

  • PDF

Evaluation of the Genetic Toxicity of Synthetic Chemicals (XIV)-in vitro Chromosomal Aberration Assay with 11 Chemicals in Chinese Hamster Lung Cells

  • Kim, Youn-Jung;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
    • /
    • 제2권2호
    • /
    • pp.89-96
    • /
    • 2006
  • The detection of many synthetic chemicals used in industry that may pose a genetic hazard in our environment is of great concern at present. Since these substances are not limited to the original products, and enter the environment, they have become widespread environmental pollutants, thus leading to a variety of chemicals that possibly threaten the public health. In this respect, to regulate and to evaluate the chemical hazard will be important to environment and human health. The clastogenicity of 11 synthetic chemicals was evaluated in Chinese hamster lung fibroblast cells in vitro. 1-Chloro-3-bromopropane CAS No. 109-70-6) induced chromosomal aberrations with significance at the concentration of $185.0\;{\mu}g/mL\;and\;1,600\;{\mu}g/mL$ both in the presence and absence of metabolic activation system, respectively. Triphenyl phosphite (CAS No. 101-02-0), which is one of the most cytotoxic chemical among 11 chemicals tested revealed no clastogenicity in the range of $95.0-4.9\;{\mu}g/mL$ both in the presence and absence of metabolic activation system. From the results of chromosomal aberration assay with 11 synthetic chemicals in Chinese hamster lung cells in vitro, 1-chloro-3-bromopropane revealed a positive clastogenic result in this study.

In vitro 검출 시스템을 이용한 한약재 열수 및 에탄올 추출물의 남성호르몬 유사활성 검출 (Detection of Testosterone-like Activities of Hydrothermal and Ethanol Extracts of Oriental Medicines Using an in Vitro Detection System)

  • 이동근;조정권;이상현
    • 생명과학회지
    • /
    • 제30권6호
    • /
    • pp.550-554
    • /
    • 2020
  • 이 연구의 목적은 시험관내 TCD (테스토스테론 화합물 검출) 시스템으로 8개 한약재의 테스토스테론 유사 활성을 검색하는 것이었다. 녹각상, 백수오, 야관문, 홍삼, 토령, 민들레, 남가세, 호로파의 열수 추출물과 80% 에탄올 추출물을 5, 50, 500 ㎍/ml의 농도로 in vitro TCD 시스템에 적용하였다. 80% 에탄올 추출물의 테스토스테론 유사 활성은 일반적으로 농도에 비례하였고, 음성대조군보다 0.9 내지 5.0 배 더 높았다. 호로파와 야관문은 동일한 농도에서 우수한 테스토스테론 유사 활성을 보였으며 민들레와 홍삼이 그 뒤를 이었다. 토령과 남가세는 낮은 테스토스테론 유사 활성을 보였으며 녹각상과 백수오는 매우 낮은 활성을 나타냈다. 양성대조군 5α-DHT의 활성과 비교하면, 호로파 및 야관문의 80% 에탄올 추출물은 500 ㎍/ml에서 10-8 M 5α-DHT보다 높은 테스토스테론 유사 활성을 나타냈다. 열수 추출물은 80% 에탄올 추출물보다 낮은 테스토스테론 유사 활성을 보였고, 농도에 따른 활성의 증가도 에탄올 추출물보다 낮았다. 호로파, 야관문 및 민들레 등의 80% 에탄올 추출물이 테스토스테론 관련 유전자의 전사 효율을 향상시킬 수 있음이 확인되었다. 따라서, 본 연구는 남성 갱년기 증상의 개선에 필요한 건강 기능성 식품 재료 또는 의약 재료를 개발하는 데 참고가 될 수 있을 것이다.