사상자 에탄올 추출물의 지방세포 분화 억제 효과

남건희(Gun He Nam),위지향(Ji-Hyang Wee),김상용(Sang Yung Kim),백지영(Ji-Young Baek),김영민(Young Min Kim) 한국생명과학회 2019 생명과학회지 Vol.29 No.9

고령화 사회에서 비만을 예방하는 것에 대한 관심이 높아지는 추세에 따라, 남성과 여성 모두 비만 관리에 상당한 비용과 노력을 지불하고 있는 실정이다. 본 연구에서는 항비만 효과를 증명하기 위해서는 3T3-L1 지방전구세포를 이용한 연구가 필수적으로 수행되기 때문에, 3T3-L1 지방전구세포에서 항비만 효과를 지닌 천연물에 조사하였다. 70% 에탄올을 이용한 사상자(Torilis Japonica Decandolle) 추출물이 3T3-L1 지방전구세포에서 성숙한 지방세포로의 분화에 미치는 효과를 Oil red O assay, western blot을 통해 확인하였다. 대조군에 비해 사상자 추출물의 100 μg/ml 농도에서 지방세포 분화와 세포 내 triglyceride (TG) 수준을 효과적으로 억제하였다. TG 함량 감소 메커니즘을 규명하기 위해, peroxisome-proliferatorsactivated-receptor-γ (PPARγ) 및 CCAAT enhancer-bindingproteins-α (C/EBPα), acetyl-CoA carboxylase (ACC)의 인산화 등 항비만 관련 단백질의 표현 수준을 확인하였다. 그 결과, 사상자 추출물은 PPARγ, C/EBPα, ACC 인산화 단백질의 발현 정도를 현저하게 감소시켰다. 종합하자면, 사상자 추출물이 비만 예방에 가장 효과적인 후보임을 명백하게 보여준다. 더 나아가서, 천연물인 사상자의 항비만 효과에 핵심적인 역할을 수행하는 활성 화합물을 탐색 및 분리하기 위한 추가 연구가 필요하다고 사료된다. Owing to increased interest in preventing obesity in an aging society, both men and women spend considerable amount of cost on obesity managements. In this study, we investigated the natural substances on anti-obesity activities in 3T3-L1 pre-adipocytes. Also, to improve anti-obesityeffects, research using 3T3-L1 pre-adipocytes cells is crucial. The anti-obesity effect of 70% ethanol extract from Torilis Japonica Decandolle on the differentiation of 3T3-L1 pre-adipocytes to adipocytes was investigated by suppressing adipocyte differentiation and lipid accumulation with Oil Red O assay, and western blot analysis. Compared to the control, 70% ethanol extract of Torilis Japonica Decandolle was significantly inhibited adipocyte differentiation and intracellular triglyceride (TG) level at a concentration of 100 μg/ml. To determine the mechanism of reduction in TG content, we determined the level of protein expression of obesity-related proteins, such as peroxisome-proliferatorsactivated-receptor-γ (PPARγ) and CCAAT enhancer-binding-proteins-α (C/EBPα), and Acetyl-CoA carboxylase (ACC) phosphorylation. As a results, 70% ethanol extract of Torilis Japonica Decandolle significantly decreased protein expression of PPARγ, C/EBPα and ACC phosphorylation. These results indicate that 70% ethanol extract of Torilis Japonica Decandolle is the most effective candidate for preventing obesity. However further studies will be needed to identify the active compounds that confer the anti-obesity activity of Torilis Japonica Decandolle.

겹삼잎국화 에탄올 추출물의 지방세포 분화 억제 효과

남건희(Gun He Nam),위지향(Ji-Hyang Wee),김상용(Sang Yung Kim),백지영(Ji-Young Baek),김영민(Young Min Kim) 한국생명과학회 2019 생명과학회지 Vol.29 No.10

겹삼잎국화는 한국에서 복통, 위염에 민간요법으로 사용되는 약초로 알려져있다. 하지만, 겹삼잎국화를 민간요법으로 사용되고 있으나 이 약초에 대한 연구는 거의 수행되어지지 않았다. 본 연구에서는 겹삼잎국화의 의학적 효과를 증명하기 위해 항비만 관련 연구를 수행하였다. 70% 에탄올을 이용한 겹삼잎국화 추출물의 항 비만 효과에 대한 연구를 위해 3T3-L1 지방전구세포를 사용하였으며, 지방전구세포가 지방세포로 성숙하는 것을 방지하고 이에 관련된 단백질 인자의 발현을 확인하기 위해 Oil Red O 염색법과 웨스턴 블롯, PCR을 통한 mRNA 분석이 수행되었다. DM (인슐린 칵테일)을 처리하여 지방전구세포의 성숙을 유도한 후, 겹삼잎국화 에탄올 추출물을 처리한 결과, 100 μg/ml 농도에서 지방세포 분화 억제 효과와 세포 내 트리글리세리드(TG)의 수준이 현저하게 감소한 것을 확인하였다. 또한, 지방전구세포의 성숙에 관여하는 여러가지 단백질 등의 인자 발현율 또한 확인하였다. 겹삼잎국화 에탄올 추출물로 인해 PPARγ, C/EBPα, LPL, FAS와 같은 지방 세포의 성숙을 유도하는 인자가 조절되었으며, Acetyl-CoA carboxylase, AMP-activated protein kinase (AMPK) 등과 같은 단백질의 발현도 억제하여 분자적 기전 또한 규명하였다. 결론적으로, 겹삼잎국화 에탄올 추출물은 지방전구세포의 활성에 영향을 끼치지 않으며 지방세포로의 성숙을 유도하는 인자 및 단백질의 발현을 선택적으로 조절하여 항 비만 효과를 갖는 것으로 확인되었다. Rudbeckia laciniata var. hortensis Bailey is used in home remedy for colic and gastritis in South Korea. Although Rudbeckia laciniata var. hortensis Bailey is used extensively for home remedies, no single study on its efficacy exists. In this study, we investigated the anti-obesity effects of Rudbeckia laciniata var. hortensis Bailey. The anti-obesity effect of a 70% ethanol extract from Rudbeckia laciniata var. hortensis Bailey on the differentiation of 3T3-L1 pre-adipocytes to adipocytes was investigated with an Oil Red O assay, western blot analysis, and mRNA analysis. Compared to the control (only treated with DM), the 70% ethanol extract of Rudbeckia laciniata var. hortensis Bailey significantly inhibited adipocyte differentiation and intracellular triglyceride (TG) levels at a concentration of 100 μg/ml. To determine how the TG content was reduced, we measured the level of protein and mRNA expression of obesityrelated agents, such as peroxisome proliferators-activated receptor γ (PPARγ), CCAAT/enhancer- binding protein α (C/EBPα), AMP-activated protein kinase (AMPK) phosphorylation, LPL, and FAS. As a result, the 70% ethanol extract of Rudbeckia laciniata var. hortensis Bailey significantly increased the expression of AMPK and decreased the expression of genes related to adipogenesis and fat storage, such as PPARγ, C/EBPα, LPL, and FAS.

Adequacy of the Anti-aging and Anti-wrinkle Effects of the Artemisia vulgaris Fermented Solvent Fraction

Ye Seul Park(박예슬),Gun He Nam(남건희),Kyung Jo Jo(조경조),Hye Won Kawk(곽혜원),Je-Geun Yoo(유제근),Jin Dong Jang(장진동),Sang Moon Kang(강상문),Sang Yong Kim(김상용),Young Min Kim(김영민) 한국생물공학회 2019 KSBB Journal Vol.34 No.3

Collagen is decomposed by MMP-1, an enzyme induced by transcription factor activator protein 1 (AP-1) and matrix metalloproteinase-9 (MMP-9). Action of MMPs in inflammatory response promotes inflammatory cell movement and secretion, resulting in wrinkles on the skin. After using Bacillus sp. fermentation system and water, Artemisia vulgaris was fermented to prepare different solvent fractions using water, dichloromethane, hex ane, n-butanol, and ethyl acetate. These fractions were used to assess their effects on cell survival, wound healing, MMP-1/MMP-9 and procollagen type I C-peptide (PICP) expression, and skin turnover. MTT assay showed that cell viability of each treated group was 103% to 121%, indicating that A. vulgaris fractions were not toxic compared to control (cell viability: 100%). Wound healing assay revealed that wound healing ability in each treated group was 51% to 61%. This was lower than wound healing area in the control. Using RT-PCR, inhibition of MMP-1/MMP-9 gene expression was examined. As a result, each group treated with fraction showed reduced expression of both MMP-1 and MMP-9 compared to tumor necrosis factor-α (TNF-α) treatment group. Effects on collagen biosynthesis were analyzed using a PICP ELISA Kit. The group in which Artemisia vulgaris was treated increased collagen synthesis from 141 to 262ng/mL compared to the control group. Three-dimensional cell culture revealed that each fraction could increase skin wall formation. These results suggest that each fractions has anti-aging and anti-wrinkle effects on the skin, indicating its suitability as a functional material.

새싹보리 에탄올 추출물을 이용한 항비만 효과 확인

박예슬(Ye Seul Park),남건희(Gun He Nam),조경조(Kyung Jo Jo),곽혜원(Hye Won Kawk),김명진(Myeong Jin Kim),김종태(Jong Tae Kim),장승희(Seung Hee Jang),김민정(Min Jeong Kim),김영민(Young Min Kim) 한국생물공학회 2020 KSBB Journal Vol.35 No.1

Obesity is caused by a variety of reasons, such as diet, genetics, and environmental factors. Here the effect of Barley sprout extract on the differentiation of 3T3-L1 and the expression of adipogenesis was studied. MTT assay for cell toxicity was performed and found to be non-toxic at 3T3-L1, with a survival rate of 90% or higher in Barley sprout extract at all concentrations. The results of oil red O assay, after inducing differentiation by treating the fat formation with insulin, dexamethasone, and IBMX powder-inducing substances at 3T3-L1, showed that the Barley sprout extract inhibited fat formation. The results of a triglyceride assay for the determination of quantitative inhibition of fat formation showed 42.2% and 45.7% in Barley sprout extracts of 50% and 70%, respectively. Western blot was performed to identify the proteins involved in fat cell suppression, and it was found that there was a concentration-dependent decrease in C/EBPα, PPARγ, and p-ACC proteins. Thus, Barley sprout ethanol extract regulates the expression of protein involved in inhibiting fat cells, thereby reducing the differentiation of fat cells and triglyceride production and indicating that it is effective against obesity.