CCD-986sk 세포 내 참깨 에탄올 추출물의 항주름 활성 연구

주다혜 ( Da-hye Joo ),유단희 ( Dan-hee Yoo ),이진영 ( Jin-young Lee ) 대한화장품학회 2016 대한화장품학회지 Vol.42 No.4

본 연구에서는 참깨 추출물의 주름개선 화장품 원료로의 가능성을 확인하기 위하여, 참깨의 70% 에탄올 추출물을 제조하여, 엘라스타제 저해능, 콜라게나제 저해능, matrixmetallopoteinases (MMPs)의 단백질, mRNA 발현 저해 효능을 측정하였다. Elastase와 collagenase 저해활성은 1000 μ g/mL 농도에서 각각 37.8%와 45%의 효소 활성을 억제를 나타내었다. 섬유아세포에서 참깨 에탄올 추출물의 세포 생존율을 확인한 결과 100 μ g/mL 농도에서 96%의 생존율을 보였다. 참깨 에탄올 추출물을 처리한 섬유아세포에서 matrix metalloproteinase-1 (MMP-1), matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-3 (MMP-3)의 단백질 발현 및 mRNA 발현 억제 효과를 확인한 결과 단백질 발현은 100 μ g/mL 농도에서 63%, 43%, 49%의 저해율을 나타내었고, mRNA 발현 억제는 최고농도인 100 μ g/mL에서 각각 82% 79%, 82%의 저해율을 나타내었다. 이러한 결과로 보아 참깨 70% 에탄올 추출물이 주름개선용 기능성 화장품 소재로서의 응용이 가능할 것으로 판단되었다. In order to investigate the possibility of Sesamum indicum L. (S. indicum) extract as an active ingredient for wrinkle-care cosmetics, we prepared 70% ethanolic extract of S. indicum and measured its elastase inhibitory activity and collagenase inhibitory activity. We also evaluated the effect of S. indicum extract on protein and mRNA expression of MMPs in fibroblast cell (CCD-986sk). For anti-wrinkle effects, elastase inhibition activities and collagenase inhibition activities were 37.8% and 45% at a dose of 1,000 μg/mL of S. indicum 70% ethanol extract. For a cell viability test, measured on fibroblast cell by ethanol extract of S. indicum, results showed 96% with cell viability at 100 μg/mL concentration. According to the results of western blot of ethanol extract from S. indicum the expression of the matrix metalloproteinase-1 (MMP-1), matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-3 (MMP-3) protein was decreased by 63%, 43%, 49% at 100 μg/mL concentration. Reverse transcription-polymerase chain reaction (PCR) of ethanol extract from S. indicum showed that the expression of MMP-1, MMP-2, MMP-3 mRNA was decreased by 82%, 79%, 82% at 100 μg/mL concentration. The findings suggest that 70% ethanol extract from S. indicum has potential as a cosmeceutical ingredient with anti-wrinkle effects.

황기 물 추출물의 미백 효과와 기능성 화장품으로서의 개발

주다혜 ( Da-hye Joo ),유단희 ( Dan-hee Yoo ),이수연 ( Soo-yeon Lee ),이진영 ( Jin-young Lee ) 한국미용학회 2015 한국미용학회지 Vol.21 No.5

The purpose of this study was to research the whitening effects and developing by cosmetics of the water extract from Astragalus membranceus Bunge, which is one of the most popular health-promoting herb in herbal medications. We performed tyrosinase inhibition assay, reverse transcription- polymerase chain reaction (RT-PCR) and western blot for whitening effects. Also we measured MTT assay for cell viability. The results were obtained as follows : For whitening effect, tyrosinase inhibition rate of water extract from Astragalus membranceus Bunge showed more than 44% at 1,000 μg/ml concentration. Cell toxicity effect on melanoma cells (B16F10) of water extract from Astragalus membranceus Bunge showed 81% with toxicity at 50 μg/ml concentration. So we were measured at a concentrations of 5, 10 and 50 μg/ml in all experiments involving cell. In addition, whitening related mRNAs including microphthalmia associated transcription factor (MITF), tyrosinase related protein-1 (TRP-1), tyrosinase related protein-2 (TRP-2), tyrosinase were reduced by Astragalus membranceus Bunge. We also found Astragalus membranceus Bunge transiently decreased protein kinase A (PKA) which is known to be upstream to the down regulation of MITF and tyrosinase. But phosphorylation of extracellular signal related kinase (pERK) were increased by Astragalus membranceus Bunge. These results imply that Astragalus membranceus Bunge decrease melanogenesis via ERK activation and subsequent down regulation of MITF and tyrosinase. Therefore, in this paper we would like to suggest the potent usage of Astragalus membranceus Bunge as materials of functional cosmetics by confirming whitening activity related with melanin content.

멜라노마 세포에서 가미소요산(加味逍遙散)의 MITF, TRP-1, TRP-2, Tyrosinase mRNA 발현 억제 효과

주다혜 ( Da Hye Joo ),이수연 ( Soo Yeon Lee ),유단희 ( Dan Hee Yoo ),이진영 ( Jin Young Lee ) 대한본초학회 2014 大韓本草學會誌 Vol.29 No.6

Objectives : Gamisoyo-san complex prescription were made with Angelicae Gigantis Radix, Paeoniae Radix, Atractylodes rhizome white, Hoelen, Bupleuri Radix, Moutan Cortex Radicis, Gardeniae Fructus, Zingiberis Rhizoma Crudus, Menthae Herba. The purpose of this study was to research the whitening effect of the extract from Gamisoyo-san, which is one of the used herbal complex prescription. Methods : This study investigated inhibitory effect of Gamisoyo-san in tyrosinase activity. Cell viability were performed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Then, Gamisoyo-san measured reversed-transcription-PCR for mRNA expression using B16F10 mouse melanoma cells. Results : For whitening effects, the tyrosinase inhibition effect of extract was shown to 52.4% at 5,000 ㎍/㎖ concentration. The cell viability on B16F10 melanoma cells of Gamisoyo-san extract showed higher than 75% at 1,000 ㎍/㎖ concentration. In this study, an experiment was performed by setting the non-toxic concentration range of 50, 150, 250 ㎍/㎖. The Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a positive control. The microphthalmia-associated transcription factor (MITF), tyrosinase related protein-1 (TRP-1), tyrosinase related protein-2 (TRP-2), tyrosinase mRNA expression inhibitory by reverse transcription-PCR of Gamisoyo-san extract were decreased by 95.3%, 98.8%, 96.3% and 49.5% at 250 ㎍/㎖ which the highest concentration. Conclusions : All these findings could verify that whitening effects of Gamisoyo-san extract by tyrosinase inhibitory activity and mRNA expression. The Gamisoyo-san could be used as material for functional cosmetics, such as skin whitening products.

재발성 또는 지속성 상피성 난소암에서의 Docetaxel 요법의 효용성 및 독성

주다혜 ( Da Hye Ju ),김용만 ( Yong Man Kim ),김민균 ( Min Gyun Kim ),김유진 ( Eu Gene Kim ),김대연 ( Dae Yeon Kim ),서대식 ( Dae Sik Seo ),김종혁 ( Jong Hyeok Kim ),김영탁 ( Young Tak Kim ),남주현 ( Joo Hyun Nam ) 대한산부인과학회 2007 Obstetrics & Gynecology Science Vol.50 No.11

목적: Paclitaxle-platinum 병합요법 치료 후, 재발하거나 반응이 없는 상피성 난소암에서 docetaxel 단독 및 병합 요법의 치료에 대한 효과 및 독성을 알아보고자 하였다. 연구 방법: 1989년 5월부터 2006년 12월까지 서울아산병원에서 재발성 혹은 지속성 상피성 난소암 환자 중 docetaxel 단독 요법 및 다른 항암제와의 병합 요법을 받은 40명의 환자를 대상으로 하였다. Docetaxel 요법은 docetaxel 75 mg/m2 단독 요법이나 docetaxel과 platinum (carboplatin AUC5 또는 cisplatin 75 mg/m2)을 1 일째 투여하는 병합 요법으로 하여 3~4주 간격으로 시행하였다. 치료 효과는 RECIST criteria와 CA-125 response criteria 정의에 따라 평가하였다. 약제 독성은 NCI Common Toxicity Criteria에 따라 평가하였다. 결과: 20명은 RECIST criteria를 기준으로, 20명은 CA-125 response criteria의 정의에 따라 평가하였다. 전체 반응률은 35% (14/40)이었고, 완전 반응은 11명 (27.5%)이었으며, 부분 반응은 3명 (7.5%)이었다. 40명의 재발성 난소암의 환자들의 평균 치료 반응 기간은 11.29개월 (4~20.7개월), 평균 병의 진행 기간은 6.91개월 (1~23개월)이었다. Platinum sensitivity에서 반응군에서는 38.7%, 저항군에서는 22.2%의 반응률을 보여 통계적으로 유의하지 않았으나 (p=0.776) 반응군에서 더 높은 반응률을 보였다. 한편 이전 3회 이상의 치료를 받은 군에서 그렇지 않은 군보다 통계적으로 유의하게 낮은 반응률을 보였다 (p=0.022). 가장 흔한 약제 독성은 탈모와 위장관계 독성이었으며, 가장 심각한 약제 독성은 골수 억제 반응이었다. 결론: 초회 치료로 paclitaxel-platinum 병용 요법을 사용 한 재발성 또는 지속성 상피성 난소암 환자에서 docetaxel은 효과와 독성 면에서 2차 치료약제로 고려할 만한 약제이다. Objective: The aim of this study is to determine the efficacy and toxicity of docetaxel in patients with recurrent or persistent epithelial ovarian cancer, previously treated with paclitaxel and platinum combination chemotherapy. Methods: Forty patients with recurrent or persistent epithelial ovarian cancer, had been treated with docetaxel combination chemotherapy at Asan Medical Center from May 1989 to December 2006. They received docetaxel (75 mg/m2) only or docetaxel (75 mg/m2) and platinum (carboplatin AUC5 or cisplatin 75 mg/m2) on day 1. The administration was repeated every 3 or 4 weeks. The response of patients was evaluated with CA-125 response criteria and RECIST criteria. The toxicities were defined according to the NCI common toxicity criteria. Results: Twenty patients had been evaluated by RECIST criteria and twenty patients had been evaluated by CA-125 response criteria. The overall response rate was 35% (14/40). Eleven patients were belonged to complete response (CR), and three patients were belonged to partial response (PR). The mean response duration (RD) was 11.29 months (4 to 20.7 months) and the mean time to progression (TTP) was 6.91 months (1 to 23 months). The response rate in the platinum-sensitive patients was 38.7% but in the platinum-resistant patients was 22.2%. The platinum-sensitive patients showed more favorable response rate, but that was not significant statistically. Heavily treated group, more than three prior regimens were used, had poor outcome. The common toxicities were alopecia and gastrointestinal toxicities (anorexia and nausea). Bone marrow suppression was the most serious drug toxicity, however, it was tolerable. Conclusion: The docetaxel is a considerable 2nd line chemotherapy with acceptable efficacy and toxicity in patients with recurrent or persistent epithelial ovarian cancer previously treated with paclitaxel and platinum combination chemotherapy.

참깨 추출물의 항산화 활성 및 melanoma cell (B16F10)에서 MITF, TRP-1, TRP-2, tyrosinase의 발현 저해

유단희(Dan-Hee Yoo,),주다혜(Da-Hye Joo),이진영(Jin-Young Lee) 한국생명과학회 2017 생명과학회지 Vol.27 No.3

본 연구는 참깨 70% 에탄올 추출물의 항산화 효과와 미백효과를 검증하여 나타내었다. 참깨 추출물의 전자공여능 측정실험은 1,000 μg/ml에서 71.7%의 효과를 나타냈으며, tyrosinase 저해활성 측정은 1,000 μg/ml의 농도에서 42%의 효과를 보였다. 참깨 추출물의 세포 생존율을 melanoma cell (B16F10)에서 확인하기 위하여 MTTassay를 진행하였으며, 세포 생존율을 측정한 결과, 1,000 μg/ml 농도에서 84.3%의 독성을 보였다. 이하의 세포실험에서는 세포 생존율이 90% 이상되는 농도인 500 μg/ml 이하에서 실험을 진행하였다. 참깨 추출물의 MITF, TRP-1, TRP-2 및 tyrosinase의 단백질 발현 효과를 50, 250, 500 μg/ml 농도에서 측정하였으며, 그 결과 MITF, TRP-1, TRP-2 및 tyrosinase의 각각 500 μg/ml 농도에서 68.3%, 39.2%, 89.7%, 22.3%의 단백질 발현 억제 효과를 보였다. 또한, MITF, TRP-1, TRP-2 및 tyrosinase의 mRNA발현을 RT-PCR로 50, 250, 500 μg/ml 농도에서 측정하였고, 양성 대조군으로 GAPDH를 사용하였다. 그 결과, 참깨 추출물의 500 μg/ml 농도에서 각각 81.8%, 66.5%, 84.2%, 68.1%의 mRNA 발현이 감소함을 확인할 수 있었다. 이상의 결과로 부터 참깨 추출물의 화장품 소재로서의 가능성을 확인할 수 있었다. This study was performed to improve the antioxidant and skin-whitening activities of 70% ethanol extract from Sesamum indicum L. (SIL). The electron-donating ability of the SIL extract was 71.7% at a concentration of 1,000 μg/ml. The whitening effects that was measured by tyrosinase inhibition assay. As a result, SIL extract was shown 42% at 1,000 μg/ml concentration. The cell toxicity on B16F10 melanoma cells of SIL of 70% ethanol extract showed 84.3% at 1,000 μg/ml concentration. The microphthalmia-associated transcription factor (MITF), tyrosinase relate protein-1 (TRP-1), tyrosinase relate protein-2 (TRP-2) and Tyrosinase protein and mRNA expression inhibitory effect of SIL extract were measured by western blot and reverse transcription- polymerase chain reaction (PCR) at 50, 250, 500 μg/ml concentration. Consequently, the MITF, TRP-1, TRP-2, Tyrosinase protein expression inhibitory effect of SIL extract was decreased by 68.3%, 39.2%, 89.7%, 22.3%, respectively, at 500 μg/ml concentration. Moreover, MITF, TRP-1, TRP-2, Tyrosinase mRNA expression inhibitory effect by reverse- transcription-PCR of SIL extract was decreased by 81.8%, 66.5%, 84.2%, 68.1%, respectively, at 500 μg/ml concentration. Therefore, we excellently identified the antioxidant activities and whitening effect of SIL extract, and this finding suggested that SIL extract has great potential as a cosmetic ingredients.

RAW 264.7을 이용한 정금나무 열매(Vaccinum oldhami fruit)의 항염증 효과

이진영(Jin-Young Lee),주다혜(Da- Hye Joo),유단희(Dan-Hee Yoo),채정우(Jung-Woo Chae) 한국생명과학회 2017 생명과학회지 Vol.27 No.4

본 연구는 천연물 소재인 정금나무 열매(Vaccinum oldhami fruit) 에탄올 추출물을 이용하여 항염증 효과에 대하여 검증하였다. 대식세포인 RAW 264.7 세포를 이용하여 정금나무 열매 추출물로 처리했을 때 세포 생존율을 확인한 결과 500 μg/ml 농도에서 118%를 나타내었다. 대식세포를 이용하여 정금나무 열매 추출물의 농도가 증가함에 따라서 변화하는 NO 생성량을 측정한 결과 최고농도 1,000 μg/ml에서 47.3%로 감소한 것을 확인할 수 있었다. 정금나무 열매 에탄올 추출물을 Western blot을 이용하여 단백질 발현을 측정한 결과 처리한 세포군에서 농도가 증가함에 따라 iNOS와 COX-2의 단백질 발현양이 감소하여 최고 농도인 500 μg/ml에서 각각 36.13%, 29.61%의 발현 억제를 보여주었다. Reverse-transcription PCR을 통하여 mRNA 발현양을 측정한 결과 iNOS와 COX-2의 mRNA 발현양이 감소하여 500 μg/ml 농도에서 62.25%, 90.07%로 내었다. 이러한 결과로 보아 정금나무 열매 에탄올 추출물의 항염증에 관한 그 효능을 확인할 수 있었고, 따라서 정금나무 열매 추출물을 이용한 천연 항염증화장품 소재개발 이용 가능성을 넓힐 수 있을 것으로 사료된다. The purpose of this study was to investigate the role of the Vaccinum oldhami fruit extract as a cosmetic additive. As a result of having macrophage (RAW 264.7) measured a cell toxicity effects of 70% ethanol extract from Vaccinum oldhami fruit, it shown 118% with toxicity at 500 μg/ml concentration. In nitric oxide synthesis inhibition effect, 70% ethanol extracts from Vaccinum oldhami fruit shown 47.3% at 1,000 μg/ml concentration. The iNOS, COX-2 protein expression inhibitory effect by western blot of 70% ethanol extract from Vaccinum oldhami fruit was decreased by 36.13%, 29.61% at 500 μg/ml concentration. And iNOS, COX-2 mRNA expression inhibitory effect by reverse-transcription-PCR of 70% ethanol extract from Vaccinum oldhami fruit was decreased by 62.25%, 90.07% at 500 μg/ml concentration. All these finding that extract from Vaccinum oldhami fruit could prove that their have effects anti-inflammatory efficacy. And extract from Vaccinum oldhami fruit has potential as a cosmetic ingredients.

연잎 추출물의 항산화 활성 및 멜라노마 세포(B16F10)에서 MITF, TRP-1, TRP-2, tyrosinase의 발현 저해 효과

유단희(Dan-Hee Yoo),주다혜(Da-Hye Joo),이수연(Soo-Yeon Lee),이진영(Jin-Young Lee) 한국생명과학회 2015 생명과학회지 Vol.25 No.10

본 연구는 연잎 추출물의 미백 화장품 첨가물로서 사용이 가능한지를 연구하였다. 연잎 추출물의 항산화 활성을 측정하게 위해 전자공여능 측정, xanthine oxidase 억제 효과 실험을 실시하였고, 미백활성을 알아보기 위하여 tyrosinase 저해활성을 측정하여 1,000 μg/ml의 농도에서 42.7%의 효과를 나타내었다. 또한 연잎 추출물에 대한 세포생존율을 MTT assay로 측정한 결과 1,000 μg/ml 농도에서 81.61%를 이상의 세포생존율을 확인할 수 있었다. 미백 관련 인자인 MITF, TRP-1, TRP-2 및 tyrosinase의 단백질 발현과 mRNA 발현 억제를 25, 50, 100 μg/ml 농도에서 측정하였다. MITF, TRP-1, TRP-2 및 tyrosinase의 단백질 발현은 100 μg/ml 농도에서 각각 69.6%, 27.7%, 67.3%, 67.8%의 저해 효과를 나타내었고, MITF, TRP-1, TRP-2 및 tyrosinase의 mRNA발현은 100 μg/ml 농도에서 각각 67.5%, 71.4%, 85.7%, 83.6%의 억제를 나타내었다. 이러한 연구결과를 보았을 때, 연잎 추출물이 항산화 및 미백활성에 효과를 나타내었고, 화장품 첨가물로서의 가능성을 확인할 수 있었다. The purpose of this study was to investigate the potential of Nelumbo nucifera G. leaf (NNL) extract as a cosmetic additive. The electron-donating ability of the NNL extract at a concentration of 1,000 μg/ml was 67.83%. In xanthine oxidase, the inhibition effect of the NNL extract was 92.7% at the same concentration. For whitening effects, tyrosinase inhibition effect of NNL extract was 42.7% at a 1,000 μg/ml concentration. The cell toxicity of the NNL extract was examined in melanoma cells (B16F10) using a 3-[4, 5–dimethyl–thiazol–2–yl]-2, 5-diphenyl-tetrazoliumbromide (MTT) assay. The cell toxicity assay revealed that the NNL extract had a toxicity of 81.61% at a concentration of 1,000 μg/ml The microphthalmia-associated transcription factor (MITF), tyrosinase related protein-1 (TRP-1), tyrosinase related protein-2 (TRP-2), and tyrosinase protein expression inhibitory effect by Western blot of NNL extract were measured by a Western blot at concentrations of 25, 50, and 100 μg/ml. At a 100 μg/ml concentration of the NNL extract, the expression of the MITF, TRP-1, TRP-2, and tyrosinase protein was decreased by 69.59%, 27.74%, 67.33%, and 67.78% respectively. The MITF, TRP-1, TRP-2 and tyrosinase mRNA expression inhibitory effect were measured by reverse transcription- polymerase chain reaction (PCR) at concentrations of 25, 50, and 100 μg/ml. GAPDH was used as a positive control. At a concentration of 100 μg/ml of the NNL extract, the expression of MITF, TRP-1, TRP-2, and tyrosinase mRNA was decreased by 67.51%, 71.36%, 85.74%, and 83.64%, respectively. These findings suggest that the NNL extract has antioxidant and whitening effects and that it has great potential as a cosmetic ingredient.

채진목 에탄올 추출물의 항염증 효과 검증

이진영 ( Jin-young Lee ),유단희 ( Dan-hee Yoo ),주다혜 ( Da Hye Joo ),김소라 ( So-ra Kim ),조희선 ( Hui-seon Jo ),주성현 ( Sung-hyun Joo ),채정우 ( Jung-woo Chae ) 대한화장품학회 2017 대한화장품학회지 Vol.43 No.1

연구에서는 채진목의 항염증 효과를 알아보기 위하여 LPS로 염증을 유도한 RAW 264.7 세포에 대한 채진목 70% 에탄올 추출물의 효과를 살펴보았다. 채진목 70% 에탄올 추출물의 대식세포에서의 세포 독성 측정을 MTT assay를 수행하였다. 세포 독성을 측정한 결과, 1,000 μ g/mL의 농도에서 96%의 세포 생존율을 나타내었다. 항염증 활성을 효과적으로 검증하기 위해, LPS로 유도된 대식세포 내 NO 생산을 억제하는 효과를 Griess의 방법으로 조사하였다. 그 결과, 채진목 70% 에탄올 추출물에서 NO의 생성이 농도 의존적으로 저해되었음을 확인하였다. 채진목 70% 에탄올 추출물을 western blot을 이용하여 단백질 발현을 측정한 결과 처리한 세포군에서 농도가 증가함에 따라 iNOS와 COX-2의 단백질 발현양이 감소하여 최고 농도인 500 μ g/mL에서 각각 84.3%, 56.2%의 발현 억제를 보여주었다. Reverse transcription-polymerase chain reaction (RT-PCR)을 통하여 mRNA 발현양을 측정한 결과 iNOS와 COX-2의 mRNA 발현양이 감소하여 500 μ g/mL 농도에서 89.8%, 84.9%로 나타내었다. 이러한 결과로 보아 채진목 70% 에탄올 추출물의 항염증에 관한 그 효능을 확인할 수 있었고, 따라서 채진목 70% 에탄올 추출물이 천연 항염증 소재로써 가능성이 있다고 판단된다. This study investigated the anti-inflammatory activities and cell viability of Amelanchier asiatica (A. asiatica) 70% ethanol extracts against RAW 264.7 cells induced by lipopolysaccharide (LPS). Cell toxicity test on macrophage cells (RAW 264.7) was performed by 3-[4,5-dimethyl-thiazol-2-yl]-2, 5-diphenyl-tetrazoliumbromide (MTT) assay and results showed 96% cell viability at 1,000 μg/mL concentration. Anti-inflammatory activity was examined via the inhibitory tests on the production of LPS induced NO in RAW 264.7 cells by Griess assay. The result showed that the extract inhibited NO production in concentration dependent manner. The iNOS and COX-2 protein expression inhibitory effects were confirmed by western blot and by reverse transcription-polymerase chain reaction (RT-PCR). From the former they were decreased by 84.3%, 56.2% at 500 μg/mL concentration, respectively, and from the latter decreased by 89.8%, 84.9% at 500 μg/mL, respectively. In conclusion, this study showed the anti-inflammatory effects of A. asiatica extracts. Thus, this could be applied to an anti-inflammatory agent.

Raw 264.7 세포에서 섬바디나물 추출물의 iNOS, COX-2 단백질 및 mRNA 발현 억제 효과

이진영 ( Jin-young Lee ),유단희 ( Dan-hee Yoo ),주다혜 ( Da-hye Joo ),채정우 ( Jung-woo Chae ) 한국미생물생명공학회(구 한국산업미생물학회) 2016 한국미생물·생명공학회지 Vol.44 No.4

본 연구에서는 섬바디 나물의 항염증 효과를 알아보기 위하여 LPS로 염증을 유도한 Raw 264.7 세포에 대한 섬바디 나물 80% 에탄올 추출물의 효과를 살펴보았다. 섬바디 나물 추출물을 LPS로 유도된 Raw 264.7 대식세포에서 전염증성 인자(iNOS, COX-2)들을 생성하여 측정하였다. 섬바디 나물 추출물의 대식세포에서의 세포 독성 측정을 MTT를 수행하였다. 섬바디 나물 추출물의 세포 독성을 측정한 결과, 1,000 μg/ml의 농도에서 100% 이상의 세포 생존율을 보였다. 섬바디 나물 추출물의 50, 100, 500 μg/ml 농도에서 iNOS와 COX-2의 단백질 발현 억제 효과를 측정하기 위해 western blot을 통해 측정하였고, 양성대조군으로는 β-actin을 사용하였다. 그 결과, 섬바디 나물 추출물을 western blot을 통해 측정한 iNOS, COX-2의 단백질 발현 억제 효과는 500 μg/ml 농도에서 각각 56%, 61.6%로 감소하였다. 섬바디 나물 추출물의 50, 100, 500 μg/ml 농도에서 iNOS, COX-2의 mRNA의 발현억제 효과를 측정하기 위해 RT-PCR을 통해 측정하였으며, 양성 대조군으로 GAPDH를 사용하였다. 그 결과, 섬바디 나물 추출물을 RT-PCR을 통해 iNOS, COX-2의 mRNA 발현 억제 효과를 측정한 결과는 500 μg/ml에서 각각 77.9%, 83.3%로 감소하였다. 이를 통해, 섬바디 나물 추출물은 염증을 억제시켜 주는 가능성이 있는 항염증 물질로써의 효과가 있을 것으로 보여진다. In this study, the anti-inflammatory activities of the 80% ethanol extract of Dystaenia takeshimana (DT) were investigated using Raw 264.7 cells treated with lipopolysaccharide (LPS). The effect of DT extract on the production of pro-inflammatory factors (iNOS, COX-2) in LPS-stimulated Raw 264.7 macrophages was examined. The cytotoxic effect of DT extract on macrophage cells (Raw 264.7) was examined by the 3-[4, 5- dimethyl-thiazol-2-yl]-2, 5-diphenyl-tetrazoliumbromide (MTT) assay. Treatment with DT extract showed 100% or more cell viability at the concentration 1,000 μg/ml. The inhibitory effect of DT extract on protein expression of inducible NOS (iNOS) and cyclooxygenase-2 (COX-2) was measured by western blotting using the concentrations 50, 100, and 500 μg/ml, with β-actin used as the positive control. Consequently, the protein expression of iNOS, and COX-2 as observed by western blotting, was decreased by 56%, 61.6%, respectively with 500 μg/ml DT extract. Inhibition of iNOS and COX-2 mRNA expression was measured by reverse transcription- polymerase chain reaction (PCR) using DT extract concentrations 50, 100, and 500 μg/ml, with GAPDH used as a positive control. Consequently, the mRNA expression of iNOS and COX-2 as observed by reverse-transcription-PCR was decreased by 77.9% and 83.3%, respectively at 500 μg/ml concentration of DT extract. In conclusion, DT extract may affect inflammatory factors as a potential anti-inflammatory agent.

산약의 멜라노마 세포(B16F10)에서 MITF, TRP-1, TRP-2, Tyrosinase, PKA, ERK 발현 억제 효과

이수연 ( Soo-yeon Lee ),유단희 ( Dan-hee Yoo ),주다혜 ( Da-hye Joo ),이진영 ( Jin-young Lee ) 대한본초학회 2015 大韓本草學會誌 Vol.30 No.4

Objectives: The purpose of this study was to research the whitening effects and developing by cosmetics of the extract from Dioscoreae Rhizoma, which is one of the most popular health-promoting herb in herbal medications. Methods: We performed tyrosinase inhibition assay, reverse transcription-polymerase chain reaction (RT-PCR) and western blot for whitening effects. Also we measured MTT assay for cell viability. Results: The results were obtained as follows : For whitening effect, tyrosinase inhibition rate of extract from Dioscoreae Rhizoma showed more than 42.28% at 1,000 ㎍/㎖concentration. Cell toxicity effect on melanoma cells (B16F10) of extract from Dioscoreae Rhizoma showed 81.97% with toxicity at 50 ㎍/㎖concentration. So we were measured at a concentrations of 5, 10 and 50 ㎍/㎖in all experiments involving cell. In addition, whitening related mRNAs including microphthalmia associated transcription factor (MITF), tyrosinase related protein-1 (TRP-1), tyrosinase related protein-2 (TRP-2), tyrosinase were reduced by Dioscoreae Rhizoma. We also found Dioscoreae Rhizoma transiently decreased protein kinase A (PKA) which is known to be upstream to the down regulation of MITF and tyrosinase. But phosphorylation of extracellular signal related kinase (pERK) were increased by Dioscoreae Rhizoma. These results imply that Dioscoreae Rhizoma decrease melanogenesis via ERK activation and subsequent down regulation of MITF and tyrosinase. Conclusions: Therefore, all these findings suggested the potent usage of Dioscoreae Rhizoma as materials of functional cosmetics by confirming whitening activity related with melanin content.