사람 섬유아세포에서 UVB 조사에 대한 능소화 추출물의 항산화 효과

김진화,이범천,표형배,Kim, Jin-Hwa,Lee, Bum-Chun,Zhang, Yong-He,Pyo, Hyeong-Bae 대한약학회 2005 약학회지 Vol.49 No.2

The human skin is constantly exposed to environmental irritants such as ultraviolet, smoke, chemicals. Free radicals and reactive oxygen species (ROS) caused by these environmen tal facts play critical roles in cellular damage. These irritants are in themselves damaging to the skin structure but they also participate the immensely complex inflammatory reaction. The purpose of this study was to investigate the skin cell protective effect of Campsis grandiflora extract on the UVB-irradiated human dermal fibroblasts (HDFs). We tested free radical and superoxide scavenging effect in vitro. C. grandiflora extracts had potent radical scavenging effect by 82% at $100{\mu}g/ml$, respectively. For testing intracellular ROS scavenging activity the cultured HDFs were analyzed by increase in DCF fluorescence upon exposure to UVB 20 $MJ/cm^2$ after treatment of C.grandiflora extracts. The results showed that oxidation of CM-DCFDA was inhibited by C.grandiflora extracts effectively and C.grandiflora extracts has a potent free radical scavenging activity in UVB- irradiated HDFs. In ROS imaging using confocal microscope we visualized DCF fluorescence in HDFs directly. In conclusion, our results suggest that C.grandiflora can be effectively used for the prevention of UV-induced adverse skin reactions such as radical production, and skin cell damage.

Tensolin-F^® (3,9-diferuloyl-6-oxopterocarpen)의 단회 독성시험 및 4주 반복투여 독성시험

김근수,박성민,이남진,표형배,채희열,정유리,임춘매,김선희,이혜영,강종구,Kim, Keun-Su,Park, Sung-Min,Lee, Nam-Jin,Pyo, Hyeong-Bae,Chai, Hee-Yul,Jung, Yu-Ri,Lin, Chun-Mai,Kim, Sun-Hee,Lee, Hye-Young,Kang, Jong-Koo 한국독성학회 2007 Toxicological Research Vol.23 No.4

This study was to investigate single and repeated-dose toxicities of Tensolin-$F^{(R)}$, an anti-wrinkle agent, in Sprague-Dawley (SD) rats or ICR mice. In single-dose oral toxicity study, the test materials were administered once by gavage to male and female SD rats at dose levels of 0 and 2,000 mg/kg. No dead animals and abnormal necropsy findings were found in control and Tensolin-$F^{(R)}$ treated group. Therefore, the approximate lethal dose of Tensolin-$F^{(R)}$ was considered to be higher than 2,000 mg/kg in rats. In the 4-week repeated oral toxicity study, the test material was administered once daily by gavage to male and female ICR mice at dose levels of 0, 25, 50 and 100 mg/kg/day for 4-weeks. In the results, no abnormality was observed in mortality, clinical findings, body weight changes, food and water consumptions, opthalmoscopic findings, necropsy findings, histopathological findings. In hematological analysis, there was a trend of increase in reticulocyte at male 25 mg/kg, although such changes were in normal ranges. On the other hand, there was a trend of decrease in hemoglobin at female 50, 100 mg/kg, such changes were in normal ranges. In addition, serum biochemical parameters including sodium, BUN and chloride increased at 25, 50 and 100 mg/kg. Relative organ weights of right testis, brain, lung and left epididymis were increased in 100 mg/kg groups of male rats in contrast to not change in female groups. However, these changes of relative organ weights, hematological and serum biochemical parameters were not accompanied with related signs such as histopathological changes or clinical findings. In conclusion, 4-week repeated oral dose of Tensolin-$F^{(R)}$ to ICR mice did not cause apparent toxicological change at the dose of 25, 50, 100 mg/kg body weight. Consequently the no-observed-adverse-effect level (NOAEL) for Tensolin-$F^{(R)}$ in ICR mice following gavage for at least 4-week is higher than 100 mg/kg/day.

새로운 미백물질을 함유하는 SLN의 제조와 봉입효율에 관한 연구

강기춘 ( Ki-choon Kang ),표형배 ( Hyeong-bae Pyo ),이청희 ( Cheong-hee Lee ),김태훈 ( Tae-hoon Kim ),마상철 ( Sang-chol Ma ),임원철 ( Noh-hee Jeong ),정노희 대한화장품학회 2008 대한화장품학회지 Vol.34 No.1

본 연구는 유용성 물질 포집에 효과적인 solid lipid nanoparticle (SLN)을 이용하여 수용성물질인 LA-PEG을 새로운 제조방법에 응용하여 실험하였다. 지질로 사용된 오일은 coconut oil, macadamia oil, 그리고 jojoba oil 3가지로 이들의 특징은 생분해성이 강하다. 외부유화제로는 Tween 20, Tween 60을 이용하여 T-SLN을 제조하였으며, SLN의 입자 분포를 비교 분석한 결과 coconut oil을 지질로 하여 사용한 것이 크기가 가장 작았으며 사용한 계면활성제의 양에 따라 입자크기와 분포형태가 달라졌다. 1 %의 Tween 60과 macadamia oil을 이용한 베이스가 입자크기가 가장 컸다. 방출관찰결과 coconut oil을 지질로 한 2 %의 Tween 20의 베이스가 가장 늦게 방출되었고, 가장 빠른 방출한 것은 Tween 60 2 % 베이스였다. This study is on encapsulation of a new whitening agent, LA-PEG using solid lipid nanoparticle (SLN) method, one of nanoparticle preparation method. Classical method has high capsulation efficiency for hydrophobic compounds but has demerit of low capsulation efficiency (2 ∼ 3 %) for hydrophilic compounds. Purpose of this study is preparation of SLN that has higher skin penetration effect compared with general liposome, and also has higher encapsulation efficiency of hydrophilic compounds. For SLN preparation, coconut oil, macadamia oil, and jojoba oil were used. As a result, SLN preparation using coconut oil (include LA-PEG) has the most high encapsulation efficiency and also has the smallest average particle size (270 nm). SLN prepared with macadamia oil and 1 % of Tween 60 has the largest particle size. Base made with coconut oil and 2 % of Tween 60 showed the fastest release and base made with macadamia oil and 2 % of Tween 20 showed the latest release.

잎새버섯 균사체를 이용한 감초추출발효물의 플라보노이드 생성과 항염 활성 연구

배준태 ( Jun Tae Bae ),송민현 ( Min Hyeon Song ),김진화 ( Jin Hwa Kim ),이근수 ( Geun Soo Lee ),표형배 ( Hyeong Bae Pyo ) 대한화장품학회 2012 대한화장품학회지 Vol.38 No.4

리퀴리티게닌과 이소리퀴리티게닌은 감초의 주요 플라보노이드 성분이다. 이들 플라보노이드는 수용성 감초 추출물과 β-glucosidase를 생성하는 잎새버섯 HB0071 균사체 발효배양을 통하여 생산하였다. 감초추출물 내 리퀴리티게닌과 이소리퀴리티게닌은 잎새버섯 발효배양 동안 현저히 증가하였다. 이 균주의 β-glucosidase의 활성은 배양 96시간을 기준으로 최고 91.5 mU/mL로 확인되었으며, 감초추출발효물로부터 생성된 리퀴리티게닌과 이소리퀴리티게닌의 함량은 HPLC 분석을 통하여 최대 568.5 μg/mL과 89.6 μg/mL로 확인되었다. 본 연구에서는 감초추출물의 잎새버섯 발효 전·후의 시료가 처리된 각질형성세포를 이용하여 자외선 UVB에 조사로 발현된 염증유발인자(COX-2)와 사이토카인(IL-1β, IL-6) 모두 감초추출발효물(FLEx)에서 농도의존적으로 발현이 억제되는 것을 확인하였다. 결론적으로 리퀴리티게닌과 이소리퀴리티게닌의 함량이 증가된 감초추출발효물은 자외선으로부터 손상된 피부 염증반응을 완화시켜줄 것으로 사료된다. Liquiritigenin and isoliquiritigenin are the major flavonoids present in licorice. These flavonoid compounds were prepared by submerged culture of Grifola frondosa (G. frondosa) HB0071 mycelia producing β-glucosidase in the aqueous extract of licorice. The contents of liquiritigenin and isoliquiritigenin were increased during the fermentation. This fungus produced a high β-glucosidase (activity of 91.5 mU/mL), thereby achieving high amounts of liquiritigenin and isoliquiritigenin (568.5 μg/mL and 89.6 μg/mL), respectively at 96 h. A reversed-phase high-performance liquid chromatography method was established for simultaneous determination of liquiritigenin and isoliquiritigenin in fermented licorice extract (FLEx). The anti-inflammatory activities were investigated by licorice extract (LEx) before and after fermentation with G. frondosa HB0071. The treatment of UVB-irradiated HaCaT keratinocytes with FLEx resulted in a dose-dependent decrease in the expression level of cyclooxygenase-2 (COX-2) mRNA. Furthermore, FLEx dose-dependently decreased mRNA of the pro-inflammatory cytokines of IL-1β and IL-6 in UVB-irradiated HaCaT cells. These results suggest that FLEx may mitigate the effects of skin inflammation by reducing UVB-induced adverse skin reactions.

가자, 오배자, 계피 추출물을 이용한 화장품 제형에서의 방부효과

조은미 ( Eun Mi Cho ),배준태 ( Jun Tae Bae ),표형배 ( Hyeong Bae Pyo ),이근수 ( Geun Su Lee ) 대한화장품학회 2008 대한화장품학회지 Vol.34 No.4

본 연구는 천연 식물 추출물을 이용하여 화장품 제형 내에서 천연 방부제로서 효능을 확인하기 위한 목적으로 진행하였다. 선별된 15종의 천연 식물 추출물을 이용하여 병원성 세균(Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli)과 효모균(Candida albicans), 곰팡이균(Aspergillus niger)에 대한 paper disc법(disc diffusion method)과 최소저해농도측정법(MIC)을 이용하여 항균 활성을 측정하였다. 그 결과 가자, 오배자 추출물은 그람 양성균인 S. aureus와 그람 음성균인 P. aeruginosa, E. coli 에 대한 항세균 활성이 우수하였고, 계피 추출물은 효모균인 C. albicans와 곰팡이균인 A. niger에 대해 우수한 항진균 활성을 나타냈다. 최종 선별된 3종 외 식물추출물은 효과가 없었다. 항균 활성 효과가 우수한 최적 조합비로 구성된 가자, 오배자, 계피 추출물의 혼합 추출물을 화장품 제형에 첨가하여 제형 내 방부 활성을 측정한 결과 상업적으로 널리 사용되고 있는 파라벤 혼합 방부제와 유사한 정도의 방부 활성을 나타냄을 확인하였다. 결론적으로, 화학 방부제를 대신하여 천연 식물 추출물로만 이루어진 넓은 항균 스펙트럼을 갖춘 방부 시스템의 개발이 가능할 것으로 사료되며 최근 다양한 문제가 제기되고 있는 화학 방부제의 단점을 극복한 천연 방부제를 함유한 화장료의 개발에 응용 가능할 것으로 기대된다. This study was carried to investigate the efficiency of antimicrobial plant extracts as natural preservative in the cosmetic formulations. Ethanol extracts of different plants were tested using the disc diffusion (paper disc) method and the minimum inhibitory concentration (MIC) method for their antimicrobial activity against the common poultry pathogens. Terminalia chebula and Rhus japonica (gallut) extracts exhibited antibacterial activity against Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli. Cinnmomum cassia extract exhibited antifungal activity against Candida albicans and Aspergillus niger while the remaining plant extracts showed no activity. A study of the preservative efficacy of the cosmetic formular containing the T. chebula, R. japonica and C. cassia extracts demonstrated sufficient preservative efficacy against bacteria and eukaryotic test microbes. Also, the cosmetic formulations containing antimicrobial plant extracts more effectively inhibited the microoranisms than the mixture of traditional chemical preservatives. These results suggest that the mixture of antimicrobial plant extracts, T. chebula, R. japonica and C. cassia is incorporated as preservative in the cosmetic formulation and the mixture have considerable effect on its efficacy.

치마버섯을 이용한 진피 발효 배양물의 항산화 및 항염 효과

송민현 ( Min Hyeon Song ),배준태 ( Jun Tae Bae ),고현주 ( Hyun Ju Ko ),장용만 ( Yong Man Jang ),이종대 ( Jong Dae Lee ),이근수 ( Geun Soo Lee ),표형배 ( Hyeong Bae Pyo ) 대한화장품학회 2011 대한화장품학회지 Vol.37 No.4

감귤은 국내 및 동남아시아 지역에서 많이 즐기는 식품 중 하나이며, 이러한 감귤류의 껍질을 건조시킨 것을 진피라고 한다. 이러한 진피는 한방에서는 이뇨작용, 비장기능을 강화하는 것으로 알려져 있으나, 천연 플라보노이드를 다량 함유하고 있어 학계에서 여러 분야로 연구가 되고 있는 실정이다. 본 연구에서는 플라보노이드 배당체를 함유한 진피 추출물을 β-glucosidase 효소를 이용하여 아글리콘(aglycone)으로 변환 할 수 있도록 치마버섯 배양을 실시하였다. 진피 발효 배양액을 HPLC를 이용하여 분석하였으며, UVA에 의한 광손상 보호능을 섬유아세포를 이용하여 측정하였다. 진피 발효 배양액은 COX-2, LOX-5와 같은 항염증 관련 대사에도 관여하여 염증완화에도 도움을 줄 수 있음을 확인하였다. 또한 각질 형성세포를 이용한 interleukin-1α를 측정한 결과 진피발효배양액 처리군의 저해활성이 더 높았다. 이러한 결과로 미루어 볼 때, 진피발효 배양액은 피부에 대한 항염 및 항산화 효과가 우수한 것으로 사료된다. Citrus unshiu (C. unshiu) Markovich were dried peel of mandarin orange, of which fresh fruit was one of the famous foods in Korea and Eastern Asia. In the oriental medicine, C. unshiu peel was known to have a diuretic effect and to strengthen spleen function. Recently, natural flavonoids of C. unshiu peel have been investigated. In this study, C. unshiu peel extract containing flavonoid-glycosides was cultured with Schizophyllum commune (S. commune) mycelia producing β-glu- cosidase and its biological activities were investigated. β-glucosidase of S. commune mycelia converted the flavonoid-glycosides (rutin and hesperidin) into aglycones (naringenin and hesperetin). Fermented C. unshiu peel extract compounds were analyzed by HPLC system. The photoprotective potential of fermented C. unshiu peel extract was tested in human dermal fibroblasts (HDFs) exposed to UVA. Fermented C. unshiu peel extract extract also showed notable in vitro anti-inflammatory effect on cellular systems generating cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX) metabolites. Also, UVB-induced production of interleukin-1α in human HaCaT cells was reduced in a dose-dependent manner by treatment with fermented C. unshiu peel extract. These results suggest that fermented C. unshiu peel extract may mitigate the effects of photoaging in skin by reducing UV-induced adverse skin reaction.

Polyglyceryl Fatty Acid Esters를 이용한 안정한 W/O 에멀젼 제조

서정민 ( Jeong Min Seo ),김형주 ( Hyeong Ju Kim ),김태훈 ( Tea Hun Kim ),강기춘 ( Gi Chun Kang ),표형배 ( Hyeong Bae Pyo ) 한국공업화학회 2008 응용화학 Vol.12 No.2

B16 Melanoma 세포에서 돌나물 추출물의 멜라닌 생성 저해 효과

심관섭,김진화,이범천,이동환,이근수,표형배,Sim, Gwan-Sub,Kim, Jin-Hwa,Lee, Bum-Chun,Lee, Dong-Hwan,Lee, Geun-Soo,Pyo, Hyeong-Bae 대한약학회 2008 약학회지 Vol.52 No.3

To develop effective skin whitening agents, we tested natural herbal extracts for their melanogenic inhibitory activities. Sedum samentosum was selected for its inhibitory effect on melanogenesis in B16 melanoma cells. Ethanolic extract of S. samentosum (SSE) was evaluated for antioxidative effect and tyrosinase inhibitory activity of melanogenesis. We investigated the changes in protein level and mRNA level of tyrosinase, tyrosinase related protein (TRP)-1 and TRP-2 by using western blotting and RT-PCR, respectively. SSE showed scavenging activities of free radicals and reactive oxygen species (ROS) with the $IC_{50}$ of 342.7 $\mug/ml$ against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and 64.69 $\mug/ml$ against superoxide radicals in the xanthine/xanthine oxidase system, respectively. SSE treatment suppressed the biosynthesis of melanin up to 46% and reduced tyrosinase activity up to 51% at 100 $\mug/ml$ in B16 melanoma cells. The tyrosinase activity and tyrosinase expression in B16 melanoma cells were reduced in a dose-dependent manner by SSE. Also, SSE was able to significantly inhibit tyrosinase and TRP-1 expression in mRNA level. These results suggest that SSE inhibited melanin production which may be dependent on tyrosinase activity and expression in B16 melanoma cells, and an effective whitening agent for the skin.

물개구리밥(Azolla imbricata)을 이용한 주요 활성성분의 분리 및 화장품 소재 개발 연구

송민현 ( Minhyeon Song ),배준태 ( Jun Tae Bae ),이근수 ( Geun Su Lee ),장영학 ( Yong He Zhang ),표형배 ( Hyeong Bae Pyo ) 대한화장품학회 2010 대한화장품학회지 Vol.36 No.1

Azolla는 Azollaceae 속에 속하는 수생 양치류이며, 물속에서 질소고정 박테리아인 Anabaena azollae와 공생관계의 식물로써, 현재까지는 질소 함량이 다량 함유되어 쌀 생산량에 크게 기여하는 식물인 것으로 알려져 있다. 이러한 이유로 인해서 중국이나 베트남 북부에서는 수생 식물의 유기 비료 뿐만 아니라 물속 정화기능까지 갖고 있는 것으로 인식되어 농업 및 수산 분야에서 연구가 활발히 진행되고 있는 실정이다. 그러나 이전 연구에서는 Azolla 속의 페놀성 화합물에 대한 연구가 미비하다. 이번 연구에서는 수생식물인 Azolla imbricata의 추출 및 분획을 통하여 Azolla imbricata fraction (AIF) 물질을 얻어 내었으며, DPPH 법을 통한 항산화능력 측정과 lipoxigenase 저해 활성 능력 측정을 통해서 Azolla imbricata의 새로운 능력을 부여하고자 하였다. 이 물질의 항산화 능력과 lipoxygenase 저해 활성능을 나타내는 물질을 분석하였으며, MMP-1 시험에서는 피부 탄력과 보호기능을 위한 콜라겐의 활성 저해율이 75 % 이상인 것으로 나타났다. LC-MASS 와 FT-IR, 원소분석(Element analysis)을 통하여 구조 분석을 하여 Azolla imbricata가 함유하고 있는 페놀성 화합물의 구조를 알아내었다. 이러한 분석을 통해서 화장품 원료로 잘 사용되지 않았던 수생식물의 사용성을 증대시키고, 천연 화장료 개발로의 응용이 가능할 것으로 기대된다. The water fern Azolla belongs to the Azollaceae and forms a symbiotic association with a N₂-fixing cyanobacterium, referred to as Anabaena azollae, and this association has currently been demonstrated to have potential as a nitrogen source for rice production. Because of that, Azolla fern has been used not only as organic manure in southern China and northern Vietnam for a long time but also as food for animate creatures in the underwater and decontaminant in the water. However, the phenolic compounds and active materials of Azolla have not been examined in detail in the past studies. In the present study, anti-oxidant ability test and experiment to find a particular active material of Azolla imbricata and Azolla imbricata fraction (AIF) were performed. In anti-oxidant test such as DPPH test and lipoxygenase inhibition test, the value of test represented high activities compared with authentic sample - green tea and NDGA (nordihydroguaiaretic acid). In MMP-1 test, related to collagen protection and elasticity of skin, its inhibitory effect was measured over 75 %, and the phenolic compounds of AIF related with this activity were confirmed luteolin derivatives by using FT-IR spectroscopy, element analyzer (EA) and Liquid chromatography-MASS spectroscopy.

잎새버섯이 생산하는 세포외 다당체의 사람 섬유아세포에서 Matrix Metalloproteinase-1 발현저해 효과

심관섭 ( Gwan Sub Sim ),배준태 ( Jun Tae Bae ),이동환 ( Dong Hwan Lee ),김진화 ( Jin Hwa Kim ),이범천 ( Bum Chun Lee ),최태부 ( Tae Boo Choe ),표형배 ( Hyeong Bae Pyo ) 대한화장품학회 2005 대한화장품학회지 Vol.31 No.2

잎새버섯(Grifola frondosa HB0071)이 생산하는 세포외 다당체인 GF-glucan이 사람섬유아세포에서 자외선(UVA)조사 시 증가되는 MMP-1 발현에 미치는 영향을 조사하였다. 자외선으로부터 조사된 섬유아세포에 GF-glucan을 처리한 농도에 따라 MMP-1 발현이 억제되었으며, RT-PCR를 이용해 세포내 MMP-1 mRNA 발현 또한 감소하는 것으로 나타났다 즉, 최대 GF-glucan 0.5%를 처리했을 때 54.4%의 MMP-1 발현을 억제하였다. 결과적으로, 잎새버섯 HB0071로부터 생산된 GF-glucan은 피부노화와 관련된 extracellular matrix (ECM) 조직에 손상을 주는 MMP-1의 활성을 억제하여 자외선으로부터 손상된 피부의 광노화로부터 보호해주는 것으로 밝혀졌다. We investigated the effect on inhibition of matrix metalloproteinase (MMP) in human dermal fibroblast (HDF) by production of exopolysaccharide (GF-glucan) from mycelial culture of Grifola frondosa HB0071. The photoprotective potential of GF-glucan was tested in HDF exposed to ultraviolet-A (UVA) light. It was revealed that GF-glucan had an inhibitory effect on MMP-1 expression in UVA-irradiated HDF without any significant cytotoxicity. The treatment of UVA-irradiated HDF with GF-glucan resulted in a dose-dependent degrease in the expression level of MMP-1 protein and mRNA (by maximum 54.4% at an 0.5% GF-glucan). These results suggest that GF-glucan obtained from mycelial culture of G. frondosa HB0071 may contribute to inhibitory action in photoaging by reducing the MMP-1 related matrix degradation system.