본 연구에서는 만성질환의 주요 위험인자인 비만을 예방 하기 위한 식품소재로서 3T3-L1세포를 이용하여 탈피한 도 토리의 항비만 효과를 알아보고자 하였다. 3T3-L1 세포에서 생존율(MTT assay)을 측정한 결과, AE와 AW 시료 모두 500 μg/mL 농도에서는 다소 생존율의 감소를 보여, 300 μg/mL를 최종 농도로 정하였다. 3T3-L1 세포의 지질축적 억제 효과를 측정한 결과, 농도 100 μg/mL로 처리하였을 때 두 시료 모두 지질축적량의 증가를 보였으나, 200 μg/mL 처리농도에서 AE 시료는 82%로, AW 시료는 74%로 감소되다가 300 μg/mL 농 도에서는 두 시료 모두 약 53% 수준까지 지질축적이 억제되 었다. 3T3-L1 세포에서 중성지방 억제 효과를 확인한 결과, AE 시료의 경우 200 μg/mL 농도에서 11%의 감소율, 300 μg/mL 농도에서 42% 수준의 감소율을 보였다. AW 시료도 200 μg/ mL 농도에서 5%의 감소율과 300 μg/mL에서 41%의 감소율 을 보였다. 3T3-L1 세포의 ROS 생성량을 측정한 결과, 시료 농도 200 μg/mL에서 AE는 42%, AW는 33%로 300 μg/mL에 서는 AE는 58%, AW는 52%로 ROS 생성량의 억제를 보였다. 3T3-L1 세포에서 mRNA 발현에 미치는 영향을 대조군과 비 교하였을 때 두 시료(AE와 AW) 모두 300 μg/mL 농도에서 PPAR-γ은 54%와 38%, aP2는 40%와 18% 수준의 발현을 억 제시키는 것으로 나타났다. 이상의 결과로 볼 때 탈피한 도토 리는 3T3-L1 세포의 분화를 억제함으로써 새로운 항비만 소 재로의 가능성이 있는 것으로 판단된다.

The aim of this study was to investigate the suppressive activity of 70% ethanol extract and its dichloromethane fraction from Auricularia auricula-judae against adipogenesis and lipogenesis in 3T3-L1 preadipocytes. The ethanol extract and its dichloromethane fraction suppressed the differentiation and decreased lipid droplets in vitro. The dose-dependent increasing concentration of glycerol and lower triglycerides accumulation were found significantly (P < 0.05) with the treatment of both fractions from 70% ethanolic Auricularia auricula-judae extract and the glycerol-3-3phosphate dehydrogenase (GPDH) activity was also inhibited by both extracts. Further, the expression of adipogenic mRNAs were investigated by RT-PCR amplification. The key transcriptional factors, PPARγ and C/EBPα were decreased significantly at dose-dependent manner by both extracts of Auricularia auricula-judae. The expressions of LPL and FAS were also decreased by presence of these extracts. The decreased expressions of C/EBPβ, C/EBPγ and ACC1 were observed only by ethanol extract at 300 μg/ml concentration, while the expression of SREBP-1c, GLUT4 and aP2 were not altered in the 3T3-L1 adipocytes. These changes were occurred without the cytotoxic effect of both extracts against 3T3-L1 adipocytes in vitro. A positive control, fenofibrate inhibited the differentiation, triglycerides accumulation through PPARγ signaling by 2/3 reduction of PPARγ expression. Thus, these findings suggest that both extracts of Auricularia auricula-judae might be used to inhibit the differentiation of 3T3-L1 preadipocytes and reduction of triglycerides accumulation.

천연물 유래 단일 성분의 지방세포로의 분화 및 지방축적 억제효과에 대한 잠재성을 탐색하기 위해 본 연구에서는 화살나무의 껍질에서 분리 정제한 FG가 3T3-L1 지방전구세포에 있어서 지방세포로의 분화 및 지방구 축적에 미치는 영향에 대해 평가하고, 관련 메카니즘에 대해 검토하였다. FG는 분화용 배지에 포함된 분화 촉진인자들의 자극에 의한 지방생성에 있어 주요한 전사인자인 PPARγ와 그 표적 단백질인 FABP4의 발현을 억제함으로써 지방세포 분화 및 지방구 축적을 억제하는 것으로 나타났다. 이상의 결과로부터 FG가 비만이나 비만과 관련된 당뇨병 등을 예방, 개선하는데 유용한 물질로 사용될 가능성이 있을 것으로 생각된다.

In the present study, we investigated the effect of 70% EtOH extract from Hippophae Rhamnoides L. leaves (HRL) on the anti-obesity effect in 3T3-L1 cells. The effects of HRL on lipid accumulation in 3T3-L1 cells were examined using Oil Red O staining. In addition, we examined the gene expression levels by using RT-PCR and western blot. The results of this analysis showed that 100 ㎍/㎖ HRL significantly increased the inhibition of lipid accumulation by 82.25%; significantly decreased the mRNA expression of sterol regulatory element binding protein-1c (SREBP-1c), peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer binding proteins α (C/EBPα), and fatty acid synthase (FAS) in 3T3-L1 cells as well as the stimulated protein expression of AMP-activated protein kinase (AMPK); and suppressed the expression level of PPARγ. These results suggest that HRL can prevent adipogenesis through activation of AMPKα and inhibition of adipogenesis transcription factors.

This study was carried out to investigate the effects of Salvia plebeia R. Br. ethanolic extract with differentaspects (stem/leaf and whole plant) on differentiation and lipid accumulation in 3T3-L1 preadipocytes. The morphologicalchanges and the degrees of lipid accumulation in 3T3-L1 cells were measured by Oil Red O staining and intra-cellular trig-lyceride (TG) assay. The mRNA expressions of special peroxisome proliferation activated receptor- genes (PPAR), CCAAT/enhancer-binding protein (C/EBPα), fatty acid synthase (FAS) and lipoprotein lipase (LPL) were detected by reverse tran-scriptase polymerase chain reaction (RT-PCR). The 50% ethanolic extracts (100μg/mL) of stem and leaf (SALE) and 30%ethanolic extracts (100g/mL) of whole plant (SAE) from Salvia plebeia R. Br. were significantly attenuated lipid accumula-tion during adipogenesis in 3T3-L1 cells. Ethyl acetate-soluble fractions (50μg/mL) significantly inhibited lipid dropletaccumulation in 3T3-L1 cells. In addition, SALE induced down-regulation of specific adipogenic transcriptional factors (C/EBPα and PPARγ) and target genes (FAS and LPL) during adipogenesis. Salvia plebeia R. Br. may be used as a safe and effi-cient natural substance to manage obesity.

Obesity, a strong risk factor for the development of chronic diseases, is characterized by an increase in the number and size of adipocytes differentiated from precursor cells, preadipocytes. Recent research suggests that increased reactive oxygen species (ROS) production in 3T3-L1 adipocyte facilitates adipocyte differentiation and fat accumulation. This study was to investigate whether reduced ROS production by Sargassum micracanthum extract (SME) could protect the development of obesity through inhibition of adipogenesis. 3T3-L1 preadipocytes were treated SME for up to 8 days following standard induction of differentiation. The extent of differentiation reflected by amount of lipid accumulation and ROS production was determined by Oil red O staining and nitroblue tetrazolium (NBT) assay. Treatment of SME significantly inhibited ROS production and adipocyte differentiation that is depend on down regulation of NADPH oxidase 4 (NOX4), a major ROS generator, and peroxisome proliferator-activated receptor gamma (PPARγ) and CCAAT/enhancer-binding protein alpha (C/EBPα), a key adipogenic transcription factor. These results indicate that SME can inhibit adipogenesis through a reduced ROS level that involves down-regulation of NOX4 expression or via modulation of adipogenic transcription factor.