Comparative Depigmentation Effects of Resveratrol and Its Two Methyl Analogues in a-Melanocyte Stimulating Hormone-triggered B16/F10 Murine Melanoma Cells

윤훈석,현창구,이남호,박성수,신동범 한국식품영양과학회 2016 Preventive Nutrition and Food Science Vol.21 No.2

Previous research showed that resveratrol (trans-3,4’,5-trihydroxystilbene) and pinostilbene (trans-3-methoxy-4’,5-dihydroxystilbene) were able to inhibit tyrosinase directly; however, anti-melanogenic effects of pterostilbene (trans-3,5-dimethoxy-4’-hydroxystilbene) and resveratrol trimethyl ether (RTE) have not been compared. To investigate the hypopigmentation effects of pterostilbene and RTE, melanin contents and intracellular tyrosinase activity were determined by western blot analysis. Firstly, pterostilbene showed the inhibitory effects on a-melanocyte stimulating hormone (MSH)-induced melanin synthesis stronger than RTE, resveratrol, and arbutin. Pterostilbene inhibited melanin biosynthesis in a dose-dependent manner in a-MSH-stimulated B16/F10 murine melanoma cells. Specifically, melanin content and intracellular tyrosinase activity were inhibited by 63% and 58%, respectively, in response to treatment with 10 mM of pterostilbene. The results of western blot analysis indicated that pterostilbene induced downregulation of tyrosinase protein expression and suppression of a-MSH-stimulated melan-A protein expression stronger than RTE or resveratrol. Based on these results, our study suggests that pterostilbene can induce hypopigmentation effects more effectively than resveratrol and RTE, and it functions via downregulation of protein expression associated with hyprepigmentation in a-MSH-triggered B16/F10 murine melanoma cells.

Differential Effects of Methoxylated p-Coumaric Acids on Melanoma in B16/F10 Cells

윤훈석,이남호,현창구,신동범 한국식품영양과학회 2015 Preventive Nutrition and Food Science Vol.20 No.1

As an approach to search for chemopreventive agents, we tested p-coumaric acid, 3-methoxy-p-coumaric acid (ferulic acid), and 3,5-dimethoxy-p-coumaric acid (sinapic acid) in B16/F10 melanoma cells. Intracellular melanin contents were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay and cytotoxicity of the compounds were examined by lactate dehydrogenase (LDH) release. p-Coumaric acid showed inhibitory effect on melanogenesis, but ferulic acid increased melanin content, and sinapic acid had almost no effect on melanogenesis. Treatment with ferulic acid resulted in a 2 to 3 fold elevation in the production of melanin. Correlatively, cell viability decreased in a dose-dependent manner when treated with ferulic acid. However, ferulic acid did not affect the LDH release from the cells. Treatment with sinapic acid resulted in a 50∼60% elevation in the release of LDH when treated with a 200 mg/mL concentration and showed neither cytostasis nor increase of melanin synthesis in a dose-dependent manner. Taken together, p-coumaric acid inhibits melanogenesis, ferulic acid induces melanogenesis, and sinapic acid exerts cytotoxic effects in B16/F10 murine melanoma cells. The results indicate that the addition of methoxy groups to p-coumaric acid shows the melanogenic or cytotoxic effects in melanoma cells compared to the original compound. Therefore, this study suggests the possibility that methoxylated p-coumaric acid, ferulic acid can be used as a chemopreventive agent.

B16/F10 생쥐 흑색종 세포에서 미역쇠 추출물의 멜라닌 생성 저해 효과

윤훈석,고원범,정재훈,김인중 제주대학교 아열대농업생명과학연구소 2009 아열대농업생명과학연구지 Vol.25 No.1

B16/F10 생쥐 흑색종 세포에서 제주조릿대 추출물의 멜라닌 합성 저해 효과

윤훈석,김정국,김세재,Yoon, Hoon-Seok,Kim, Jeong-Kook,Kim, Se-Jae 한국생명과학회 2007 생명과학회지 Vol.17 No.6

Effects of hot-water extract from Sasa quelpaertensis leaf (HWES) on melanogenesis were investigated in B16/F10 mouse melanoma cells. HWES inhibited cellular tyrosinase activity and melanin biosynthesis in a dose-dependent manner. Western blotting analysis showed that HWES dose-dependently inhibited tyrosinase and tyrosinase related protein-1 expression. Also, HWES suppressed sustained ERK activation in a concentration-dependent manner, suggesting that HWES inhibits the melanin biosynthesis through the suppressive effect against pathway involving sustained ERK activation.

백합뿌리 추출물의 멜라닌 생성 억제효과

윤훈석(Hoon Seok Yoon),양경월(Kyung-Wol Yang),김정은(Jung Eun Kim),김정미(Jeong Mi Kim),이남호(Nam Ho Lee),현창구(Chang-Gu Hyun) 한국식품영양과학회 2014 한국식품영양과학회지 Vol.43 No.5

피부미백 소재를 개발하기 위해 멜라닌 함량, 세포내 tyrosinase 활성의 측정 및 Western blotting 실험이 수행되었다. 백합(Lilium Oriental Hybrid ‘Siberia’) 뿌리의 80% 에탄올 추출물로부터 얻은 에틸아세테이트 분획물(R-EA)은 α-melanocyte stimulating hormone(α-MSH)에 의해 멜라닌 생성이 유도된 B16/F10 흑색종 세포에서 농도 의존적으로 멜라닌 생성을 저해하였다. 정확하게 세포내 tyrosinase 활성과 멜라닌 함량은 에틸아세데이트 분획물 100μg/mL 처리 시 α-MSH 단독 처리군에 비해 각각 45%와 74%의 저해율을 보였다. α-MSH에 의해 멜라닌 생성이 유도된 B16/F10 흑색종 세포에서 단백질 발현양상을 살펴본 결과 TRP-1이 가장 많이 억제된 양상을 확인할 수 있었고 이 결과는 세포내 tyrosinase 활성저해보다 멜라닌 생성저해가 더 많이 일어난 것과 일맥상통하는 것이다. 이를 종합해 볼 때 p-coumaric acid와 resveratrol 함량이 백합뿌리의 에탄올 추출물에 비해 많이 함유된 에틸아세테이트 분획물은 멜라닌 생성 유도물질에 의해 촉진된 ERK의 활성화를 억제하는 피부미백 소재로서 그 가치가 입증된다고 사료된다. In order to develop a skin-whitening agent, melanin contents and intracellular tyrosinase activity were determined by western blotting. Ethyl acetate fractions of 80% ethanol extracts from lily (Lilium Oriental Hybrid ‘Siberia’) bulbs (R-EA) inhibited melanin synthesis in a dose-dependent manner in α-melanocyte stimulating hormone (α-MSH)-treated B16/F10 murine melanoma cells. Intracellular tyrosinase activity and melanin contents were suppressed by 45% and 74%, respectively, in response to treatment with 100 μg/mL of R-EA. Examination of protein expression associated with α-MSH-induced melanogenesis showed that tyrosinase related protein (TRP)-1 was inhibited more strongly than tyrosinase, and these results were correlated with stronger inhibition of melanin synthesis than intracellular tyrosinase activity. Taken together, R-EA containing p-coumaric acid and resveratrol could be used as a hypopigmentation agent through suppression of sustained extracellular signal-regulated kinase (ERK) activation via melanogenic induction.

새발 추출물의 멜라닌 생성 저해 효과

윤훈석 ( Hoon Seok Yoon ),김정국 ( Jeong Kook Kim ) 대한화장품학회 2007 대한화장품학회지 Vol.33 No.2

본 연구에서는 천연 미백소재 개발을 위하여 제주도 연안에 서식하는 해조류 중 홍조류의 일종인 새발 추출물의 멜라닌 생성에 연관된 생리활성을 분석하였다. 그 결과, 새발 추출물의 폴리페놀 함량은 2.0 % 이하였고, 라디칼 소거활성(DPPH)은 IC₅₀값이 2,000 μg/mL 이상이었으며, 세포독성은 관찰되지 않았다. 그리고 생쥐의 B16/F10 흑색종 세포에서 멜라닌 생합성에 관련된 효소의 저해효과를 알아본 결과, 알파-멜라노사이트 자극 호르몬에 의해 유도된 extracellular signal-regulated kinase 1/2 (ERK 1/2)의 활성화를 억제함으로써 티로시나제와 티로시나제 연관 단백질 1을 저해하는 것으로 확인되었다. 따라서 새발 추출물은 멜라닌 생합성에 필수적인 효소인 티로시나제의 저해활성과 티로시나제 연관 단백질 1의 발현억제 효과가 뛰어난 것으로 분석되었다. 그러므로 홍조류인 새발(Acanthopeltis japonica)의 추출물은 멜라닌 생성 자극제로 유도된 신호전달경로를 저해하는 미백 기능성 화장품 소재로서의 활용 가능성이 높을 것으로 사료된다. To develop the skin whitening agent, we investigated the effects of Acanthopeltis japonica, a rhodophyta on the coast of Jeju island, on melanogenesis. Dried A. japonica was refluxed with 70 % aqueous ethanol and the extract was evaporated to dryness. To validate the activity as a depigmenting agent, various in vitro tests, polyphenol contents, and free radical scavenging activity were performed. In addition, cellular tyrosinase activity and protein expression of p-ERT, tyrosinase, TRP-1, and TRP-2 were measured in B16/F10 murine melanoma cells. A. japonica had low polyphenol contents and low free radicals scavenging activities against 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical. A. japonica suppressed cellular tyrosinase activity up to 86.9 % at 100 μg/mL with inhibition or tyrosinase and TRP-1 expression in α-melanocyte stimulating hormone (α-MSH)-treated B16/F10 melanoma cells. Our results suggest that inhibitory effects of A. japonica on melanogenesis are due to inhibiting the pathways involving α-MSH-induced ERK activation. Therefore, A. japonica nay be useful as a skin whitening agent associated with the suppressive effect of melanotrophin-induced signaling pathway to inhibit melanin synthesis.

해양 기상상태가 선박사고에 미치는 영향: 통영 해역을 중심으로

이경준(Kyeongjun Lee),윤훈석(Hoonseok Yoon),한수현(Suhyun Han),정준호(Junho Jeong),김연재(Yeonjae Kim) 한국데이터정보과학회 2020 한국데이터정보과학회지 Vol.31 No.1

최근 선박 사고의 수는 매년 증가하였다. 해상의 선박 사고는 항해사의 운항부주의 등과 같은 선원에 의한 인적과실이 대부분을 차지하지만 해상의 기상 상황에 의해서도 영향을 받는다. 따라서 본 연구는 다양한 해양 기상 요인의 영향 정도에 따른 선박 사고 발생 여부의 분석을 목적으로 하였다. 통영 해역의 2018년도 선박 사고 발생 자료와 기상청 기상자료개방포털의 해양 기상 자료 (풍속, 대기압, 수온, 파고, 파주기), 시간대, 요일을 활용하여 로지스틱 회귀 모형과 의사결정나무 모형을 이용하여 분석하였다. 그 결과 해양 기상 요인 중 수온이 선박 사고 발생에 가장 큰 영향을 미치는 요인으로 나타났다. 이러한 결과는 해상 기상 상태에 따른 선박 사고의 발생을 예측하는데 유용하게 사용할 수 있을 것이다. Recently, boating accidents have been increased every year. The carelessness of officers, many marine weather factors have a great influence on the boating accidents. Especially, the number of marine accident is governed by wind speed, atmospheric pressure, water temperature, wave height and wave period. The purpose of this paper is to analyse the effect of marine weather factors on boating accident. We use the data of boating accident, marine weather factors (wind speed, atmospheric pressure, water temperature, wave height and wave period), time zone and day of the week in 2018. We did statistical analysis using logistic regression analysis and decision tree analysis. These prediction models may be used to predict the boating accident according to the marine weather condition.

궁천조생 감귤에서 Carotenoid Isomerase type 1과 type 2 유전자의 분리와 발현분석

김성우(Seong Woo Kim),윤훈석(Hoon Seok Yoon),김인중(In-Jung Kim) 한국원예학회 2011 한국원예학회 학술발표요지 Vol.2011 No.5

Docosahexaenoic acid의 모유두세포 증식 효능 및 기전

고지연,오일중,강정일,최윤경,윤훈석,유은숙,고창익,안용석,Ko, Jiyeon,Oh, Il-Joong,Kang, Jung-Il,Choi, Youn Kyung,Yoon, Hoon-Seok,Yoo, Eun-Sook,Ko, Chang-Ik,Ahn, Yong-Seok 제주대학교 의과학연구소 2019 The Journal of Medicine and Life Science Vol.16 No.3

Docosahexaenoic acid (DHA), a principal of mackerel-derived fermented fish oil, increases the proliferation of dermal papilla cells (DPCs) via the upregulation of cell cycle-associated proteins such as cyclin D1 and cdc2 p34, and might promote hair-growth. However, the intracellular mechanisms that underlie the action of DHA in the proliferation of DPCs have not been investigated fully. In this study, we addressed the action mechanisms of DHA to trigger the activation of anagen in DPCs. DHA activated β-catenin signaling by the increased phosphorylation at serine 552 and serine 675 as well as the translocation and accumulation of activated β-catenin into the nucleus. In the other hand, DHA inhibited canonical TGF-β/Smad signaling by the decreased phosphorylation of Smad2/3. Taken together, the results indicate that DHA might stimulate anagen signaling via the activation of Wnt/β-catenin pathway, while the inactivation of canonical TGF-β signaling pathway in DPCs.

제주 재래식된장의 생리적 특성

황준호(Joon-Ho Hwang),오유성(You-Sung Oh),임자훈(Ja-Hun Lim),박지은(Ji-Eun Park),김미보(Mi-Bo Kim),윤훈석(Hoon Seok Yoon),임상빈(Sang-Bin Lim) 한국식품영양과학회 2009 한국식품영양과학회지 Vol.38 No.12

제주재래식된장의 생리적 특성을 측정하기 위하여 제주도내 전통된장업체로부터 된장제품 4종(JTD1-4)을 수거하여, 유기용매로 추출, 분획하여 총 페놀 함량, 항산화활성(DPPH 유리기 소거능, superoxide 유리기 소거능), ACE 및 α-glucosidase 저해능, 항염 및 항암효과를 측정하였다. 에틸아세테이트 분획물에서 총 페놀 함량(201.8~305.4 ㎍/ g)과 DPPH 유리기 소거활성(37.5~59.9%)이 가장 높았다. Superoxide anion radical 소거활성은 부탄올 분획물에서 56.3~85.1%로 가장 높았는데, JTD1(81.7%)과 JTD4(85.1%)의 소거활성은 양성대조구로 사용한 ascorbic acid의 소거활성(82.2%)과 유사하였다. ACE 저해활성은 메탄올 추출물에서 23.6~74.5%로 가장 높았다. α-Glucosidase 저해활성은 디클로로메탄 분획물에서 72.0~80.2%로 양성대조구인 acarbos의 57.4%보다 높았다. RAW 264.7 세포를 대상으로한 NO 생성 저해활성(IC50: 17.3~29.0 ㎍/mL)은 에틸아세테이트 분획물이 가장 높았지만, 세포독성은 모든 분획물에서 관찰되지 않았다. 에틸아세테이트 분획물의 세포증식 저해활성은 정상세포주인 피부섬유아세포(HS-68)에서 JTD1-3은 100 ㎍/mL 이하의 농도에서 나타내지 않았으나, JTD4는 21%의 세포독성을 나타내었다. 대장암 세포주(HT-29)에 대해서는 JTD1과 JTD2는 세포증식이 저해되지 않았으나, JTD3과 JTD4는 50 ㎍/mL 이상에서 증식저해 효과를 나타내었다. 유방암 세포주(MCF-7)는 100 ㎍/mL에서 JTD3은 30.9%, JTD4는 34.7%의 세포증식 저해효과를 보였다. Four Jeju traditional Doenjangs (JTD), a traditional fermented soybean paste, were extracted and fractionated using organic solvents, and their physiological properties were evaluated such as total phenolic contents (TPC), antioxidant capacity (DPPH and superoxide free radical scavenging), inhibitory effect of ACE and α-glucosidase, and anti-inflammatory and anti-cancer activities. Ethyl acetate fractions had the highest TPC (201.8~305.4 ㎍/g) and DPPH radical scavenging activity (37.5~59.9%). Buthanol fractions had the highest superoxide anion radical scavenging activity (56.3~85.1%), and the activities of JTD1 (81.7%) and JTD4 (85.1%) were almost the same as that of ascorbic acid (82.2%) as the reference material. Inhibitory activities of ACE were 23.6~74.5% in methanol extracts, and those of α-glucosidase were 72.0~80.2% in dichloromethane fractions compared with 57.4% in acarbos as the reference. Inhibitory activities of NO production in LPS-induced RAW 264.7 were the highest as 17.3~29.0 ㎍/mL (IC50) in ethyl acetate fractions, and cell toxicities were not detected in all fractions. Ethyl acetate fractions of JTD1-3 showed no inhibitory activities, but that of JTD4 showed 21% of inhibitory activity of cell growth against human genital skin fibroblast cell line HS-68. Cell growth of human colon cancer cell line HT-29 was not inhibited by ethyl acetate fractions of JTD1 and 2, but was inhibited by those of JTD3 and 4 at more than 50 ㎍/mL. JTD3 and JTD4 showed the growth inhibition of breast adenocarcinoma cell line (MCF-7) by 30.9% and 34.7% at 100 ㎍/mL, respectively.