이사람 / ' 정도를 지키는 직장인 ' - 김한혁

김한혁 한국해사문제연구소 1993 (月刊)海洋韓國 Vol.233 No.-

국내자생식물 추출물의 항산화 및 항염 활성 탐색

김한혁,권주희,박관희,김많흔,오명환,최강인,박상인,진혜영,김성식,이민원 한국생약학회 2012 생약학회지 Vol.43 No.1

181 kinds of local native plants were selected by its anti-inflammatorial folk medicinal uses and evaluated it antioxidative and inhibitory activity of nitric oxide (NO) production in lipopolysaccharide (LPS)-induced 264.7 macrophage cells. Among the 181 kinds of plants, 99 species showed potent antioxidative activities and 20 extracts showed inhibitory activity towards nitric oxide production by more than 70% at a concentration of 100 µg/mL. Therefore, these plants should be considered promising candidates for the treatment of inflammatory diseases accompanying overproduction of NO.

Flavonoid constituents in the leaves of Myrica rubra sieb. et zucc. with anti-inflammatory activity

김한혁,Dong-Hee Kim,김많흔,오명환,김소라,박광준,이민원 대한약학회 2013 Archives of Pharmacal Research Vol.36 No.12

The leaves of Myrica rubra sieb. et zucc. havebeen used in oriental traditional medicine for the treatmentof burns, skin diseases, and as an antidiarrheal in China,Japan, and Korea. Activity guided isolation of the leaves ofM. rubra has led to the isolation of five flavonoid: myricetin(1), myricitrin (2), myricetin 3-O-(200-O-galloyl)-a-L-rhamnopyranoside (3), myricetin 3-O-(200-O-galloyl)-b-D-galactopyranoside (4), and quercetin 3-O-(200-O-galloyl)-b-D-galactopyranoside (5). All isolates were evaluatedfor their antioxidant potency against the superoxideanion (O2-), and compounds 3–5 showed potent scavengingactivities with 50 % inhibition concentration (IC50)values compared to the positive control, allopurinol. Compounds 1–5 were evaluated as inhibitors of variousmacrophage functions involved in the inflammatory process. These five compounds significantly and dose dependentlyinhibited lipopolysaccharide (LPS)-stimulated nitricoxide (NO), pro-inflammatory cytokines, and the proteinlevels of inducible nitric oxide synthase (iNOS) andcyclooxygenase-2 (COX-2) in LPS-stimulated RAW 264.7macrophages. Our results suggest that galloyl flavonolglycosides (3–5) isolated from M. rubra might be beneficialfor the treatment of inflammation-related diseases.

Inhibition of matrix metalloproteinase-1 and type-I procollagen expression by phenolic compounds isolated from the leaves of Quercus mongolica in ultraviolet-irradiated human fibroblast cells

김한혁,김동희,오명환,박광준,허준혁,이민원 대한약학회 2015 Archives of Pharmacal Research Vol.38 No.1

The aim of this study was to investigate theeffect of Quercus mongolica (QM) which induce antiphotoagingprocess of skin in vitro. Bioassay-guided isolationof 80 % Me2CO extract of the leaves of QM led tothe isolation and identification of six known phenoliccompounds: pedunculagin (1), (-)-epigallocatechin (2),(?)-catechin (3), quercetin 3-O-(600-O-galloyl)-b-D-glucopyranoside(4), kaempferol-3-O-b-D-glucopyranoside-7-Oa-L-rhamnopyranoside (5) and kaempferol 3-O-(600-galloyl)-b-D-glucopyranoside (6). The effects of compounds1–6 on expression of matrix metalloproteinase-1 (MMP-1)and type-I procollagen were further evaluated. Amongthem, compound 1 showed potent inhibitory effect onMMP-1 and the increased type-I procollagen synthesis inultraviolet B-induced human fibroblast. These results suggestthat pedunculagin, an ellagitannin, is a potential candidatefor the prevention and treatment of skin aging.

Inhibitory Effects on Melanin Production in B16 Melanoma Cells of fractions from Schisandra chinensis

김한혁,정아름,서미자 한국고분자학회 2019 한국고분자학회 학술대회 연구논문 초록집 Vol.44 No.2

Antiproliferative Effects of Native Plants on Prostate Cancer Cells

김한혁,이민원,박관희,김많흔,오명환,김소라,박광준,허준혁 한국생약학회 2013 Natural Product Sciences Vol.19 No.2

As part of the research for the natural products about prostate-related disease, this study screened 159 plant species from 46 families, which included a total of 213 different kinds of local native plants and these plants were tested for the ability to inhibit LNCaP proliferation, an androgen-sensitive prostate cancer cell line, and DU145 proliferation, which is a more aggressive androgen-insensitive prostate cancer cell line. The results indicated that nineteen of 213 types of plants exhibited antiproliferative activity (cell viability < 30%, 500 mg/mL) on the growth of androgen-sensitive LNCaP cell lines, and five of them exhibited DU145 cell antiproliferative activity (cell viability < 30%, 500 mg/mL). The methanol extracts of Eurya emarginata (stems), Gleditsia japonica var. koraiensis (leaves), Photinia glabra (leaves) and Elaeagnus macrophylla (leaves) showed antiproliferative activity on both the androgen-sensitive LNCaP cells (cell viability < 30%) and androgen-insensitive DU145 cells (cell viability > 100%). The study also found that the methanol extracts of Styrax japonica (fruits), Aralia continentalis (leaves), Fagus crenata var. multinervis (stems), Thuja orientalis (stems) and Poncirus trifoliate (branches) presented the strongest activity and demonstrated potent antiproliferative activity on both cell lines (LNCaP and DU145 cell viability < 30%).

섬쑥부쟁이(Aster glehni Fr. Schm.) 분획물의 항산화 및 항염증 활성

김한혁 ( Han Hyuk Kim ),박근혜 ( Gun Hye Park ),박강수 ( Kang Soo Park ),이진영 ( Jin Young Lee ),안봉전 ( Bong Jeun An ) 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 2010 한국미생물·생명공학회지 Vol.38 No.4

최근 연구에서 폴리페놀을 함유한 천연물은 질병의 위험과 노화의 원인인 ROS와 RNS를 감소시키는 것과 연관이 있다고 밝혔다. 본 연구에서는 섬쑥부쟁이 추출물과 분획물의 산화와 염증에 대한 저해 효과가 있는지에 대한 연구를 조사하였다. 섬쑥부쟁이 70% 메탄올 추출물은 헥산층, 에틸 아세테이트층, 부탄올층, 물층을 극성별로 분획하였다. 섬쑥부쟁이로부터 분획한 에틸아세테이트 추출물과 부탄올 추출물의 전자공여능은 50 μg/mL에서 각각 58.0%, 46.4%를 나타냈다. 에틸아세테이트 추출물의 superoxide anion radical 소거능은 50 μg/mL에서 64.65%를 나타냈고, 부탄올 추출물은 50 μg/mL의 농도에서 35.66%의 저해 효과를 나타내었다. 항염증 효과와 관련된 hyaluronidase와 lipoxygenase의 저해 활성에 대한 에틸아세테이트 분획물의 효과는 5 μg/mL의 농도에서 각각 24.37%, 29.5%의 저해 효과를 나타냈다. 이러한 에틸아세테이트 추출물의 항염증 효과는 nitric oxide 생성 억제에서도 유의했으며, Raw 264.7 cell 내 LPS에 의해 유도된 iNOS 단백질에서 50 μg/mL의 EtOAc 추출물을 처리시 81.5%의 발현을 억제하는 효과가 확인되었다. The Plants and their extracts containing polyphenol have been shown to be associated with decreased the cause of aging and variety of disease such as reaction oxygen species (ROS) and reactive nitrogen species (RNS) in several recent studies. We conducted to investigate whether the extracts and fractionation isolated from Aster glehni Fr. Schm. has an inhibitory effect association with oxidation or inflammation. The Aster glehni Fr. Schm. 70% aq. MeOH was fractioned according to polarity with n-hexane layer, EtOAc layer, n-BuOH layer and water layer. The electron donating ability of EtOAc, n-BuOH solvent fraction from Aster glehni Fr. Schm. was about 58.0%, 46.4% at 50 μg/mL, respectively. The superoxide anion radical inhibitory effect of EtOAc extracts was about 64.65% at 50 μg/mL, and n-BuOH extracts was 35.66% at 50 μg/mL. EtOAc layer to the inhibition activity of hyaluronidase and lipoxygenase were inhibited about 24.37%, 29.5% at 5 μg/mL. In the anti-inflammation effect of EtOAc layer inhibited the generation of nitric oxide. also, these results showed that EtOAc extract inhibited 81.5% at 50 μg/mL on the expressions of iNOS protein in Raw264.7 cell line.

오미자 잎 분획물의 B16F10 멜라노마 세포 내 색소침착 억제 효과

김한혁(Han-Hyuk Kim),나형진(Hyung-Jin Na),정아름(A-Leum Jeong),서미자(Mi-Ja Seo) 한국인체미용예술학회 2020 한국인체미용예술학회지 Vol.21 No.1

The Melanin synthesis is controlled by a series of enzymatic reactions regulated at the level of tyrosinase in melanocytes and melanoma cells. In an ongoing search for new tyrosinase inhibitors from natural sources, the dried leaves of Schisandra chinensis were extracted with 70% acetone and separated into fractions 1-3 (Fr.1-Fr.3). All fractions were evaluated for their inhibitory activity potency against the tyrosinase, and Fr.2 showed potent tyrosinase activities with 34.7% inhibition concentration 1,000 μg/mL compared to the positive control, albutin (23.5%). Finally, melanin contents was measured to determine the whitening effect. Fr.2 was inhibited at 19.3% at 50μg/mL, respectively. We investigated the changes in protein level of tyrosinase, Melanocyte Inducing Transcription Factor (MITF) by using western blotting. Fr.2 was able to significantly inhibit tyrosinase expression in protein level. These results suggest that Fr.2 inhibited melanin production which may be dependent on tyrosinase activity and expression in B16F10 melanoma cells, and an effective whitening agent for the skin.

마우스 흑색종세포에서 Purpurogallin의 멜라닌 생성 억제 효과

김한혁(Han-Hyuk Kim),김태훈(Tae Hoon Kim) 한국식품영양과학회 2015 한국식품영양과학회지 Vol.44 No.12

천연 미백소재 개발과 관련하여 많은 연구들이 멜라닌 합성저해 및 활성 메커니즘을 규명하는 데 초점이 맞춰졌으며, 이러한 이유로 tyrosinase 저해제 개발이 다양하게 이루어져 왔다. 본 연구는 배 유래의 polyphenol oxidase를 이용하여 천연에 존재하는 단순 폴리페놀인 pyrogallol의 산화축합반응을 유도하여 purpurogallin을 효율적으로 생합성하였으며, 본 화합물에 대해서 미백 활성을 평가하였다. 먼저 MTT assay를 통해 세포독성이 없는 농도구간을 설정하였으며, purpurogallin은 25 μM 농도의 melanoma 세포 내에서 tyrosinase 활성을 20% 이상 저해하는 것을 확인하였다. 또한 25 μM의 시험 농도에서 purpurogallin은 약 20%이상의 melanin 생합성 저해 활성을 나타내었다. 미백 관련 전사인자인 MITF, TRP-1, TRP-2, tyrosinase의 단백질 발현을 측정한 결과, 본 화합물은 B16F10 melanoma 세포에서 tyrosinase, TRP-1과 TRP-2의 단백질 생합성을 두 추출물 모두 억제하는 것을 확인하였다. Tyrosinase, TRP-1과 TRP-2의 발현을 조절하는 전사인자로는 MITF가 관여하는 것으로 알려져 있으며, 실제로 MITF는 melanin 생성과 관련된 여러 유전자의 발현을 조절하는 데 중요한 작용을 하고 있다. 따라서 purpurogallin은 melanin 생성과 관련된 중요한 세 가지 단백질의 생합성을 전사단계에서 조절 전사인자인 MITF의 단백질 발현을 억제하는 효과가 있음을 확인하였다. 이상의 결과로부터 멜라닌 생합성에 있어서 상위신호단계에 있는 전사인자 MITF의 활성을 억제함으로써 하위 신호전달 과정을 억제하는 것임을 시사하며, 향후 추가적인 검증작업을 통해 화장품 소재화가 가능할 것으로 판단된다. Melanin is one of the most important factors affecting skin color. Melanogenesis is the bioprocess of melanin production by melanocytes in the skin and hair follicles and is mediated by several enzymes, such as tyrosinase, tyrosinase related protein (TRP)-1, and TRP-2. Convenient enzymatic transformation of the simple phenol pyrogallol with polyphenol oxidase originating from pear to an oxidative product, purpurogallin, was efficient. The structure of the pyrogallol oxidation product was identified on the basis of spectroscopic methods. The biotransformation product purpurogallin showed significant inhibitory effects against both melanin synthesis and tyrosinase activity in a dose-dependent manner in B16 melanoma cells. In addition, purpurogallin significantly attenuated melanin production by inhibiting TRP-1, and TRP-2 expression through modulation of their corresponding transcription factors, and microphthalamia- associated transcription factor in B16 cells. Consequently, purpurogallin derived from convenient enzymatic transformation of pyrogallol might be a beneficial material for reducing skin hyperpigmentation.

섬쑥부쟁이(Aster glehni Fr. Schm.) 분획물의 미백 및 주름 개선 효과

Han-Hyuk Kim(김한혁),Gun-hye Park(박근혜),Kang-Soo Park(박강수),Jin-Young Lee(이진영),Tae-Hoon Kim(김태훈),Bong-Jeun An(안봉전) 한국생명과학회 2010 생명과학회지 Vol.20 No.7

천연의 식물과 추출물은 비용의 효율성과 풍부한 자원을 이점으로 식품, 약, 의약 분야와 마찬가지로 화장품 분야에서도 이용될 수 있다. 현재까지 섬쑥부쟁이는 항산화 활성을 제외하고는 그 효능에 대해 알려진 바가 없다. 따라서 본 연구에서는 울릉도의 대표적인 특산 식물인 섬쑥부쟁이 추출물을 이용하여 화장품 분야에서의 활용 방안에 대한 연구를 조사한바, 효소적 in vitro 검색법을 측정하여 이들 중 가장 우수한 EtOAc 추출물을 대상으로 이러한 저해능이 멜라닌 생성에 관련된 단백질 발현과도 연관성이 있는지를 확인하기 위해 B16F10 melanoma cell line 내 단백질 수준을 확인하였다. Tyrosinase, TRP-1의 항체를 이용한 western blot으로 관련 단백질의 발현량을 조사한 결과 섬쑥부쟁이 에틸아세테이트 분획물 100 ㎍/㎖을 처리한 군에서는 tyrosinase protein을 30.5%를 억제하였고, TRP-1은 41.5% 억제하였다. 한편, 섬쑥부쟁이 분획물의 생육 저해환 측정에서는 Staphylococcus epidermidis, Propionibacterium acnes, Escherichia coli에 대하여 헥산 분획물 2 ㎎/disc에서 각각 21.0 ㎜, 12.0 ㎜, 14.0 mm의 저해환을 나타내어 섬쑥부쟁이 헥산 분획물만이 유일하게 높은 항균활성을 나타났다. 이들 실험 결과를 미루어보아, 섬쑥부쟁이의 화장품 분야에서의 활용적 가치가 충분하다고 사료되나, 그 활성 성분에 대한 명확한 규명을 위한 추가적인 연구를 제시하는 바이다. Plants and their extracts can be utilized as inexpensive and rich resources of active constituents in the cosmetic field, as well as the food, pharmaceutical and medicinal fields. Until now, Aster glehni Fr. Schm. had no known active effect, except on anti-oxidation, that was found during investigations for application in the cosmetic field. In this study, we examined the inhibition of enzymatic reactions to protein levels in inclusive B16F10 melanoma cell lines. Significant inhibition of enzymatic reactions was observed in the EtOAc extract, which advanced to tyrosinase protein and TRP-1 in the B16F10 melanoma cell line. These results indicated that the effect of EtOAc extract inhibited expressions of tyrosinase protein and TRP-1 in the B16F10 melanoma cell line by 30.5% and 41.5% at 100㎍/㎖ respectively. On the other hand, antimicrobial activity was evaluated to the four fractions in normal flora of the skin. Hexane extract was only exhibited in the higher clear zone in all strains. In conclusion, any cosmeceutical effects of Aster glehni Fr. Schm. may have a potential meaning, as well as possible value for further studies regarding the effects of chemical constituents of Aster glehni Fr. Schm.