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인체 각질형성세포 배양모델을 이용한 수종 용제의 독성도 측정에 관한 연구
유재학 ( Jae Hak Yoo ),은희철 ( Hee Chul Eun ),정진호 ( Jin Ho Chung ),유일재 ( Il Je Yu ) 대한피부과학회 1996 대한피부과학회지 Vol.34 No.1
Background: As in vivo models such as animal and human tests have many potential problems the keratinocyte culture model has previously been used as an in vitro model for testing skin irritancy for common skin irritants. Objective : To determine the skin irritant potency of several solvents, we employed cultured human keratinocytes as an in vitro model. Methods : To evaluate the cytotoxicity of solvents, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MTT) test, lactic dehydrogenase(LDH) release test, and tritiated thymidine incorporation test were used. Results : Dose dependent decrease of cell viability and DNA synthesis, and dose dependent increase in leakage of LDH liberation were observed in normal cultured human keratinocytes after exposure to several solvents. The cytotoxicity potency of several solvents measured by each method were slightly different. Conclusion : These observations suggest that the cultured human keratinocyte model would be useful in evaluating the cytotoxicity of solvents. (Kor J Dermatol 1996;34(1): 58~63)
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비침습적 측정방법을 이용한 수종 용제가 자극 피부염에 미치는 영향
유재학(Jae Hak Yoo),은희철(Hee Chul Eun),서대헌(Dae Hun Suh),유일재(Il Je Yu) 대한피부과학회 1994 대한피부과학회지 Vol.32 No.6
Background : Solvents play an immense role in the industria sector. Irritant dermatitis which is more common than allergic contact dermatitis can be caused by solvents, Objective : Our purpose was to compare the skin irritancy of several solvents using human and guinea pig skin models. Methods : The skin responses to short contact with etharol, acetone, dimethylsulfoxide (DMSO) and xylene were measured by visual scoring of erythm, transepidermal water loss and laser doppler flowmetry. Results : The results are summarized as follows . 1. Guinea pig and human skin responses to normal saline, ethanol, and acetone were nearly negligible. 2. Guinea pig skin responses to 99.9% DMSO under occlusion for 0.5 min were assessed by visual scoring system, TEWL, and LDF. They are measured 3+0.(0,1+21.70, 45+12.70 at 5 min after removal of 99.9% DMSQ, and 0.83+0.41, 10.5+3.83, 36+4.0, to 120 min after removal. 3. Guinea pig skin responses to 97% xylene under occlusion for 5 nin were assessed by visual scoring system, TEWL and LDF. They are measured 3+0.00, 1. 5.82, 77+11.7 at 5 min after removal of 97% xylene, and 1.83+0.75, 5.5+3.21, 39.17+11.53 at l2 min after removal. 4. Human skin responses to 75% DMSO under occlusion for 1 min were assessed by visual scoring system, TEWL and LDF. They are measured 2,5+0.5, 63+25.8, 51+13.7 at 5 min after removal of 75% DMSO, and 0.17+0.41, 14.67+15.87, 21.17-8. 1 at 120 min after removal. 5. Human skin responses to 97% xylene under occlusion for 12 min were assessed by visual scoring system, TEWL and LDF. They are measured 2.7+0.52+4.22, 76+14.30 at 5 min after removal of 97% xylene, and 0+0, 2.5+0.55, 3.17+0.98, 120 emOVBI. Concbsion : Short contact vrith DMSO and xylene cause visib erythema and an increase in TEWL and cutaneous blood flow. The reaction patterns in hurr an and guinea pig skin models were similar. (Kor J Dermatol 1994: 32(6): 1026-1034)
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금은화 추출물의 항산화 효과를 통한 세포 보호효과 및 유전자 발현 양상
조원준(Won June Cho),윤희승(Hee Seung Yoon),김용현(Yong Hyun Kim),김정민(Jung Min Kim),유일재(Il Jae Yoo),한만덕(Man-Deuk Han),방인석(In Seok Bang) 한국생명과학회 2013 생명과학회지 Vol.23 No.8
금은화의 메탄올 추출에 의한 유기 용매별 분획에서 항산화 효과를 근거로 에틸 아세테이트 분획에서 H₂O₂로 유도된 RAW 264.7 세포독성에 대한 세포보호 효과와 유전자 발현 양상을 분석하였다. 용매별 분획의 항산화 활성은 시료의 농도가 증가할수록 DPPH에 대한 전자공여능도 증가하였으며, ED50은 에틸 아세테이트 분획에서 39.56 μg/ml로 가장 높게 나타났다. 또한 H2O2에 의해 유도된 Raw 264.7 세포사멸((IC50)에 대하여 에틸 아세테이트 분획은 농도 의존적으로 유의적인 세포 생존율과, 100 μg/ml의 농도에서 82.49%의 세포보호 효과를 나타내었다. 한편 IC50의 H₂O₂ 독성에 대한 에틸 아세테이트 분획의 Raw 264.7 세포보호 효과에 관련된 유전자 발현의 패턴은 BHA와 유사한 것으로 나타났다. 항산화 관련 유전자군의 비교 분석에서는 에틸 아세테이트 분획과 BHA는 매우 유사한 유전자 발현 양상을 보였으나, 세포 내 주요 항산화 효소를 만드는 유전자 중 하나인 heme oxygenase1 (Hmox1) 유전자의 발현 정도가 에틸 아세테이트 분획에서 4 배 이상 증가하였다. 항염증 관련 유전자군의 비교에서는 T-box transcription factor (Tbx21)가 BHA 처리군에서는 2 배 감소하였으나 에틸 아세테이트 분획의 처리군에서는 오히려 약 2 배 증가한 것으로 나타났다. In this study, based on the antioxidative effects in organic solvent fractions obtained from the main methanolic extract of L. japonica, the protective cellular effects and gene expression patterns of ethyl acetate fractions on H₂O₂-induced Raw 264.7 cell death (IC50) were analyzed. The antioxidant activity of the fractions measured using DPPH free radical scavenging activity increased in a dose-dependent manner, and the ED50 exhibited the highest 39.56 μg/ml in the ethyl acetate fraction. In addition, the ethyl acetate fractions’ cell viability on H₂O₂-induced Raw 264.7 cell damage increased in a concentration- dependent manner, showed a visible cell survival rate of 82.49% at a concentration of 100 μg/ml. The gene expression patterns related to the ethyl acetate fractions’ cytoprotective effect in H₂O₂-induced Raw 264.7 cell damage presented similar patterns to those of BHA. In comparative analysis for antioxidant activity-related genes affected by ethyl acetate fractions and BHA in H₂O₂-induced Raw 264.7 cells, both ethyl acetate fractions and BHA showed very similar gene expression patterns, but the gene expression level of the heme oxygenase 1 (Hmox1) gene making antioxidant enzymes in cells was four times higher in ethyl acetate fractions than BHA. In inflammation-related genes in H₂O₂induced Raw 264.7 cells, the T-box transcription factor (Tbx21) gene was expressed about two times more frequently in the ethyl acetate fraction treatment group, while it was expressed half as frequently in the BHA treatment group.
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