소사나무(Carpinus turczaninowii, C. turczaninowii) 잎 추출물의 멜라닌 생성 억제활성을 B16F10 melanoma 세포를 이용하여 측정하였다. 그 결과, 에탄올 추출물($100{\mu}g/mL$)에서 72.2%의 생성 억제 효과를 확인하였으며, 동일 농도에서 MTT 세포독성은 거의 나타나지 않았다. 분획물(헥산, 에틸아세테이트, 부탄올 및 물)을 제조한 후 실험을 진행한 결과, 에틸아세테이트 분획에서 가장 우수한 활성이 관찰되었다. 에틸아세테이트 분획에서 활성성분을 규명하기 위하여 크로마토그라피를 진행하였으며, 4개의 화합물을 분리 동정하였다; ethyl gallate (1), quercetin rhamnose (2), kaempferol rhamnose (3), quercetin galloylrhamnose (4). 화합물의 구조규명은 핵자기공명분광기 등을 이용하여 이루어졌으며, 4개의 화합물 모두 소사나무 잎에서는 처음 분리된 물질이다. 분리된 화합물을 대상으로 멜라닌 생성 억제활성 실험을 진행한 결과, 화합물 4에서 세포독성 없이 농도의존적인 억제활성을 확인하였다. 또한, 화합물 4는 세포 내에서 티로시나제 발현양을 감소시키고 있음을 ELISA를 통하여 확인하였다. 이상의 결과를 바탕으로, 소사나무 잎 추출물이 화장품에서 미백제로 활용될 가능성이 있다고 판단된다.
소사나무(Carpinus turczaninowii, C. turczaninowii) 잎 추출물의 멜라닌 생성 억제활성을 B16F10 melanoma 세포를 이용하여 측정하였다. 그 결과, 에탄올 추출물($100{\mu}g/mL$)에서 72.2%의 생성 억제 효과를 확인하였으며, 동일 농도에서 MTT 세포독성은 거의 나타나지 않았다. 분획물(헥산, 에틸아세테이트, 부탄올 및 물)을 제조한 후 실험을 진행한 결과, 에틸아세테이트 분획에서 가장 우수한 활성이 관찰되었다. 에틸아세테이트 분획에서 활성성분을 규명하기 위하여 크로마토그라피를 진행하였으며, 4개의 화합물을 분리 동정하였다; ethyl gallate (1), quercetin rhamnose (2), kaempferol rhamnose (3), quercetin galloylrhamnose (4). 화합물의 구조규명은 핵자기공명분광기 등을 이용하여 이루어졌으며, 4개의 화합물 모두 소사나무 잎에서는 처음 분리된 물질이다. 분리된 화합물을 대상으로 멜라닌 생성 억제활성 실험을 진행한 결과, 화합물 4에서 세포독성 없이 농도의존적인 억제활성을 확인하였다. 또한, 화합물 4는 세포 내에서 티로시나제 발현양을 감소시키고 있음을 ELISA를 통하여 확인하였다. 이상의 결과를 바탕으로, 소사나무 잎 추출물이 화장품에서 미백제로 활용될 가능성이 있다고 판단된다.
Melanin synthesis inhibition activities were investigated for the extracts prepared from the leaves of Carpinus turczaninowii (C. turczaninowii) by using B16F10 melanoma cells. As a result, the ethanol extract ($100{\mu}g/mL$) showed 72.2% inhibition activities without cell toxicities in ...
Melanin synthesis inhibition activities were investigated for the extracts prepared from the leaves of Carpinus turczaninowii (C. turczaninowii) by using B16F10 melanoma cells. As a result, the ethanol extract ($100{\mu}g/mL$) showed 72.2% inhibition activities without cell toxicities in MTT assays. For the solvent fractions (n-hexane, ethyl acetate, n-butanol, water), the most potent activities were observed at the ethyl acetate fraction. To isolate the active constituents, the ethyl acetate fraction was further purified to afford four compounds; ethyl gallate (1), quercetin rhamnose (2), kaempferol rhamnose (3) and quercetin galloylrhamnose (4). The identification of the isolates was made by spectroscopic data including NMR spectra, and all of the compounds 1-4 were isolated for the first time from the leaves of C. turczaninowii. Anti-melanogenesis activities were studied for the isolates 1-4, and the compound 4 was determined to decrease the melanin synthesis dose-dependently without causing cell toxicities. ELISA measurement indicated that the isolate 4 decreased the contents of cell tyrosinase, a critical enzyme in melanogenesis. Based on these results, the extracts of C. turczaninowii were found to be applicable as whitening ingredients in cosmetic formulations.
Melanin synthesis inhibition activities were investigated for the extracts prepared from the leaves of Carpinus turczaninowii (C. turczaninowii) by using B16F10 melanoma cells. As a result, the ethanol extract ($100{\mu}g/mL$) showed 72.2% inhibition activities without cell toxicities in MTT assays. For the solvent fractions (n-hexane, ethyl acetate, n-butanol, water), the most potent activities were observed at the ethyl acetate fraction. To isolate the active constituents, the ethyl acetate fraction was further purified to afford four compounds; ethyl gallate (1), quercetin rhamnose (2), kaempferol rhamnose (3) and quercetin galloylrhamnose (4). The identification of the isolates was made by spectroscopic data including NMR spectra, and all of the compounds 1-4 were isolated for the first time from the leaves of C. turczaninowii. Anti-melanogenesis activities were studied for the isolates 1-4, and the compound 4 was determined to decrease the melanin synthesis dose-dependently without causing cell toxicities. ELISA measurement indicated that the isolate 4 decreased the contents of cell tyrosinase, a critical enzyme in melanogenesis. Based on these results, the extracts of C. turczaninowii were found to be applicable as whitening ingredients in cosmetic formulations.
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제안 방법
3% inhibition of melanin production at 100 µg/mL concentration. The cell toxicities were examined for the ethanol extract and solvent fractions by using MTT test. As shown in Figure 1, cell toxicities were not detected as cell viabilities were maintained at the indicated concentrations for all of the fractions.
대상 데이터
B16F10 murine melanoma cell was purchased from the American Type Cell Culutre (ATCC, USA). These cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM, Gibco Inc.
turczaninowii were collected in May 2011 from Halla botanical garden in Jeju Island, Korea. Voucher specimen (sample number 308) was deposited at the herbarium of department of chemistry, Jeju National University.
이론/모형
The cell viability was determined by 3-(4,5-Dimethylthiazol -2-yl)-2,5-diphenyl tetrazolium bromide (MTT) cell viability assay. Briefly, cells were seeded into 6-well cell culture plates.
성능/효과
When the isolated compound 4 was examined with varying concentrations, it also showed inhibitory activities for the tyrosinase enzyme production in a dose-dependent manner (Figure 5). These data demonstrated that the melanin synthesis inhibition by the C. turczaninowii extract could be associated with the down-regulation of the tyrosinase enzyme in the melanoma cells.
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