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한국동물실험대체법학회> 동물실험대체법학회지

동물실험대체법학회지 update

Journal of Alternatives to Animal Experiments

  • : 한국동물실험대체법학회
  • : 의약학분야  >  기타(의약학)
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  • : 1975-9657
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수록정보
수록범위 : 1권1호(2007)~14권1호(2020) |수록논문 수 : 103
동물실험대체법학회지
14권1호(2020년 12월) 수록논문
최근 권호 논문
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KCI후보

1연잎 추출물과 연잎 발효 추출물의 항산화 및 항염 효과 비교

저자 : 김지은 ( Jieun Kim ) , 김배환 ( Bae-hwan Kim ) , 권수경 ( Soo Kyung Kwon )

발행기관 : 한국동물실험대체법학회 간행물 : 동물실험대체법학회지 14권 1호 발행 연도 : 2020 페이지 : pp. 5-14 (10 pages)

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This study tended to identify the potential as functional natural materials for soothing of skin and scalp using lotus leaf hot water extract (Nelumbo nucifera hot water extract, NHWE) and fermented lotus leaf extract (fermented nelumbo nucifera leaf extract, FNLE).
The antioxidant evaluations such as total polyphenol and flavonoid content, DPPH electron-donating ability, and ABTS radical scavenging ability, were measured. Cell survival rate was measured in Raw 264.7 cells treated with LPS to check the toxicity, and the inhibitory rate of NO-producing and TNF-α and COX-2 protein expression were measured in Raw 264.7 cells after treatment of LPS in order to evaluate the efficacy of anti-inflammatory effect.
NHWE and FNLE showed 182.2 mg/g and 103.3 mg/g each in total polyphenol contents, and 26.8 mg/g and 49.1 mg/g each in flavonoid content. As for ABTS radical scavenging ability, high levels of antioxidants (86.6% and 93.7% in NHWE and FNLE) appeared at the concentration of 1000 μg/mL. In DPPH electron-donating ability, NHWE and FNLE showed 66.1% and 73.3% at 1000 μg/mL. FNLE had more antioxidant ability than NHWE. In anti-inflammatory effects, inhibition rate of NO production by LPS increased in a concentration-dependent manner at both NHWE and FNLE, and TNF-α and COX-2 expressions were also decreased by treatment of NHWE and FNLE. In particular, 200 μg/mL of FNLE showed the best anti-inflammatory effect.
In summary, fermented lotus leaf extract showed good efficacy in antioxidant and anti-inflammatory tests, and its potential as a functional ingredients was confirmed. It is expected that not only the soothing effect of the scalp and skin, but also the possibility of application as a functional food can be expanded, and it is considered to be suitable as a natural functional raw material.

KCI후보

2피부감작능 평가 대체시험법간 위양성 및 위음성 화학물질에 대한 인체 세포주 활성화방법의 전수에 따른 예측력 평가

저자 : 조아랑 ( Ahrang Cho ) , 여경욱 ( Kyenguk Yeo ) , 정미숙 ( Misook Jung ) , 이재희 ( Jaehee Lee ) , 양수정 ( Sujeong Yang ) , Ravi Gautam , 이기용 ( Giyong Lee ) , 조지훈 ( Jihun Jo ) , Manju Acharya , Anju Maharjan , 김창열 ( Changyul Kim ) , 허용 ( Yong Heo )

발행기관 : 한국동물실험대체법학회 간행물 : 동물실험대체법학회지 14권 1호 발행 연도 : 2020 페이지 : pp. 15-21 (7 pages)

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The in vitro alternative assay for evaluating skin sensitizing potentials (SSP) called human Cell Line Activation Test (h-CLAT) has been recently adopted as an international test guideline. The present study was proceeded to pursue the domestic establishment and expansion of h-CLAT. The standard operational procedure was prepared in detail by the lead laboratory (LL) and transferred to one participating laboratory (PL). The LL demonstrated technical proficiency for the 10 substances listed in OECD TG 442E. The PL also demonstrated a certain level of proficiency, in that 4-phenylenediamine and 4-aminobenzoic acid distributed with coded names were correctly classified as positive and negative on SSP, respectively, and other indices including 75% cell viability (CV75) and relative fluorescence intensity (RFI) % for CD54 and CD86 were fell within the reference range. Methyl methacrylate, nickel chloride, and resorcinol, which are inconsistent in predicting SSP through various alternative test methods, were unanimously predicted as negative, positive, and positive on SSP by the two laboratories for the first time through h-CLAT. Both chlorobenzene and sodium lauryl sulfate with the existing h-CLAT report as positive and negative, respectively, were predicted as negative by the PL, which could contribute toward overall categorization into non-skin sensitizer.

KCI후보

3Profiling on Altered Productions of Cytokines or Chemokines from THP-1 Cell Line Following in Vitro Exposure to Cyclophosphamide, Cyclosporine, Dexamethasone, or Tacrolimus, Representative Immunosuppressants

저자 : Manju Acharya , Jihun Jo , Anju Maharjan , Ravi Gautam , Daeun Lee , Changyul Kim , Yong Heo

발행기관 : 한국동물실험대체법학회 간행물 : 동물실험대체법학회지 14권 1호 발행 연도 : 2020 페이지 : pp. 23-30 (8 pages)

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Although immune suppressive drugs have contributed for the cancer chemotherapy, organ transplant, but their therapeutic efficacy is limited. This study aims to evaluate the immunological modulations induced by four representative immunosuppressive on THP-1 cell line. 75% cell viabilities were determined and four test concentrations, 0.01X, 0.1X, 0.5X, and vehicle control were chosen. Culture supernatants were collected at 24 h after lipopolysaccharide (LPS1 ㎍/㎖) activation in the presence of test substances. 27 target cytokines were measured through luminex system and relative cytokine production levels (RCPLs,%) were calculated. Cytokines with the RCPL below 100% at all the three concentrations were 21 including IL-1beta, -1ra, -2, -4, -5, -6, -7, -9, -10, -12, -13, -15, -17, Eotaxin, FGF-basic, GM-CSF, IFNγ, IP-10, MCP-1α, PDGF-BB, and VEGF in dexamethasone-treated cells. Concerning on cyclophosphamide, the RCPLs on 11 cytokines were less than 100% at all the 3 concentrations. Treatment of cyclosporine demonstrated 10 cytokines with below 100% RCPL at all the 3 concentrations. The RCPLs below 100% were observed with 22 cytokines including IL-1β, -1ra, -2, -4, -5, -6, -7, -9, -12, -13, -15, -17, Eotaxin, GFG-basic, G-CSF, GM-CSF, IFNγ, MCP-1, MIP-1α, PDGF-BB, TNFα, and VEGF from THP-1 cells treated with tacrolimus at all the 3 concentrations. The present study indicates that tacrolimus and dexamethasone are stronger immunosuppressants than cyclophosphamide and cyclosporine on THP-1 cell line.

KCI후보

4두 가지 단계적 접근법을 이용한 30종 화학물질의 안자극성 평가

저자 : 전혜련 ( Hye Lyun Jeon ) , 조아랑 ( Ah Rang Cho ) , 고경육 ( Kyung Yuk Ko ) , 김주환 ( Joohwan Kim ) , 정미경 ( Mi Kyung Jeong ) , 박교현 ( Kyo-hyun Park ) , 김배환 ( Bae-hwan Kim ) , 오원준 ( Won Jun Oh ) , 이종권 ( Jong Kwon Lee ) , 박기숙 ( Ki Sook Park )

발행기관 : 한국동물실험대체법학회 간행물 : 동물실험대체법학회지 14권 1호 발행 연도 : 2020 페이지 : pp. 31-39 (9 pages)

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Various alternative test methods are being developed to replace in vivo Draize rabbit test that evaluates eye irritation. However, a single alternative method has difficulty to be applied in safety evaluation on substances, because it cannot fully replace the in vivo test by simulating only part of the in vivo system. For this reason, different studies using several alternative test methods and test results of literatures have been under way. Our previous study suggested effective tiered approaches using three tests among Short Time Exposure (STE), Bovine Corneal Opacity and Permeability (BCOP), Hen's Egg Test-Chorioallantoic Membrane (HET-CAM), re-constructed human cornea epitheliums (RhCE) tests. This study aimed to evaluate eye irritation potential for 30 new test chemicals using two tiered approaches. According to the data generated by direct test performance and literature survey, the accuracy, sensitivity and specificity of the two tiered approaches were 83.3%∼86.7%, 93.3%, 73.3%∼ 80%, respectively. Furthermore, the accuracy, sensitivity, and specificity of the two tiered approaches by applying combined data against 30 test chemicals in our present study and 47 test chemicals in previous study were 90.9%∼92.2%, 95.9%∼98.0%, 78.6%∼85.7%. Consequently, the two tiered approaches may be used to identify between irritants and non-irritants to replace in vivo test.

KCI후보

5치올기 함유 저분자 화합물과의 반응성을 이용하는 간단한 in chemico 피부감작성 시험법의 개발 가능성 평가

저자 : 김건호 ( Geon Ho Kim ) , 차동호 ( Dong Ho Cha ) , 구형모 ( Hyung Mo Gu ) , 박재성 ( Jae Sung Park ) , 정재훈 ( Jae Hoon Jeong ) , 황진선 ( Jin Sum Hwang ) , 네팔마헤시 ( Mahesh Raj Nepal ) , 정태천 ( Tae Cheon Jeong )

발행기관 : 한국동물실험대체법학회 간행물 : 동물실험대체법학회지 14권 1호 발행 연도 : 2020 페이지 : pp. 41-48 (8 pages)

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For the initiation of skin sensitization reaction, a sensitizer should react with endogenous proteins. During the haptenization of a sensitizer with proteins, it is also possible for a sensitizer to react with small molecules that contain either amino or thiol group, indicating a possibility to develop a simple skin sensitization test using the reactivity of test compounds with thiol-containing low molecular weight compounds. In the present study, 10 compounds that contain a thiol group were pre-tested to determine whether the reactivity with a thiol-detecting monobromobimane would be in concentration-dependent manners. Subsequently, among thiol compounds, a simple spectrofluorometrical method to identify skin sensitizers in chemico was tested with glutathione, an endogenous substance that contains amino and thiol groups. To flurorometrically quantitate the remaining thiol-containing compounds following a reaction with skin sensitizers, monobromobimane was employed. The conditions optimized included: incubation conditions of thiol compounds including glutathione with monobromobimane, molar ratios of thiol compounds to monobromobimane, optimal concentration and incubation time of monobromobimane, and pH and incubation time with test compounds. With a tentatively optimized conditions, 4 skin sensitizers and 2 non-sensitizers were tested to know whether the method could correctly identify skin sensitizers and non-sensitizers. The results indicated a good possibility. Although further optimization is required, it would be a useful screening tool for determining skin sensitization potential of test compounds, because the present method employs a simple endogenous thiol compound as an acceptor for sensitizers with a spectrofluorometric detection system requiring neither experimental animals nor cell cultures.

KCI후보

6RAW 264.7 cell에서의 미모사 에탄올 추출물의 항산화 및 항염증 효능 연구

저자 : 전민영 ( Min Yeong Jeon ) , 김배환 ( Bae-hwan Kim )

발행기관 : 한국동물실험대체법학회 간행물 : 동물실험대체법학회지 14권 1호 발행 연도 : 2020 페이지 : pp. 49-57 (9 pages)

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The purpose of this study is to identify the possibility of Mimosa pudica ethanol extracts (MEE) as a cosmeceutical material with antioxidant and anti-inflammatory effects.
At first, the antioxidant effect of the MEE was carried out through Total polyphenol, 2,2diphenyl-1- picrylhydrazyl (DPPH) electro-donating activity, 2,2'-azino-bis diammonium salt (ABTS+) radial scavenging activity. The total polyphenol content of the MEE was 3.514 mg/g. DPPH electro-donating activity of MEE were increased in a dose-dependent manner (P<0.05), and was about 88.1% at 1 mg/mL concentration. Also, ABTS+ radical scavenging activity of MEE were remarkably increased in a dose-dependent manner (P<0.05), and was about 99.7% at 1 mg/mL concentration. The MEE showed relatively high antioxidant effects in most experiments.
To evaluate the anti-inflammatory effects in RAW 264.7 cells, the maximum permissible concentration was verified first by cell viability assessment using MTT assay. And then, nitric oxide (NO) production inhibition and amounts of inflammatory related proteins and gene expressions were identified after LPS treatment. NO inhibition of MEE was decreased concentration dependently, and was about 84.3% at 1 mg/mL concentration. And TNF-α and IL-6 proteins by western blotting were also decreased significantly by treatment of MEE. As a result of measuring the expression levels of TNF-α and IL-6, the expression inhibition rates were 67.87% and 71.91%, respectively.
Based on the results of these experiments, we suggest that MEE could be useful as a cosmeceutical material with anti-inflammatory effects.

KCI후보

7Mulberroside A 및 Oxyresveratrol의 병용 처리에 의한 미백 시너지 효과

저자 : 박희진 ( Heui-jin Park ) , 허윤정 ( Yoon-jung Huh ) , 임경민 ( Kyung-min Lim )

발행기관 : 한국동물실험대체법학회 간행물 : 동물실험대체법학회지 14권 1호 발행 연도 : 2020 페이지 : pp. 59-65 (7 pages)

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Melasma is an acquired hyper-melanosis characterized by irregular light to dark brown macules and patches on sun-exposed areas of the skin. Topical treatments of melasma involve the use of hypo-pigmenting agents which modifies various stages of melanogenesis. The most common mode of action for hypo-pigmentation is the inhibition of the tyrosinase. Mulberroside A and oxyresveratrol, well known tyrosinase inhibitors, are found frequently together in natural products. The aim of the experiment is to find out the synergism in decreasing pigmentation by mulberroside A and oxyresveratrol. B16F10 murine melanoma cells were used for identification of melanin content and cell viability. In addition, inhibition against tyrosinase, a key enzyme for melanin synthesis, was determined. Single treatment of mulberroside A or oxyresveratrol exhibited dose-dependent inhibition of mushroom tyrosinase activity, which showed a reduction of extra melanin in B16F10 melanoma cells. Interestingly, while mulberroside A or oxyresveratrol rarely reduced melanin inside the melanocytes alone, their mixture could significantly attenuate the production of intracellular melanin, which might be manifested as synergistic hypopigmenting effects. Further studies are needed to understand the mechanism of synergism by mulberroside A and oxyresveratrol in the inhibition of melanin synthesis.

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