본 연구는 십자화과 채소인 갓(Brassica juncea) 추출물 을 이용하여 지표성분인 sinigrin의 함량을 분석하고, 내분비계 교란물질 환경호르몬인 비스페놀 A (BPA)로 분화를 유도한 3T3-L1 전구지방세포에서 갓 추출물과 sinigrin 처 리에 대한 지방세포 분화 및 활성산소종(ROS) 생성 억제, 지방 생성 전사인자(PPARγ, C/EBPα, aP2)의 단백질 발현 감소 효능을 평가하였다. 연구 결과에 따르면 HPLC를 이용하여 측정한 갓 추출물 중 sinigrin의 함량은 21.27±0.2 mg/g 인 것으로 나타났다. BPA로 유도된 3T3-L1 전구지방세포에 서 XTT assay 결과 sinigrin 180 μM 및 갓 추출물 300 μg/ mL 농도에서 세포 독성을 보이지 않았으며, 지방세포 분화과정 중 세포 내 지방 축적량과 ROS 생성량을 비교하였을 때 갓과 sinigrin을 처리한 지방세포의 경우 지방축 적량 및 ROS 생성량 모두 유의적으로 감소시키는 것으로 나타났다. 또한, 갓 추출물 및 sinigrin을 처리하였을 때 지 방세포 분화를 조절하는 전사인자 PPARγ, C/EBPα 및 aP2 의 발현이 억제됨을 확인하였다. 이 결과를 통해 갓은 환 경호르몬 비스페놀 A로 유도된 지방세포 내 지방 축적 억제와 더불어 ROS 저감에 효과적으로 작용함을 확인하였다. 향후 sinigrin을 함유한 갓은 BPA로 인한 지질 대사 장애를 예방하는 천연물 유래 기능성 식품 소재로 활용할 가능성이 높은 것으로 기대되며, 추가로 임상 연구 및 작용기전 입증을 위한 in vivo 모델에서의 후속 연구가 진 행되어야 할 것으로 사료된다.
(-)-Epigallocatechin-3-gallate (EGCG) is a major catechin found in green tea. It is reported that EGCG possesses various health benefits including anti-cancer, antioxidant, anti-diabetes, and anti-obesity. The objective of this study was to investigate the effects of EGCG on adipogenesis via activation of AMP-activated protein kinase (AMPK) pathway in 3T3-L1 preadipocytes. In order to determine the effects of EGCG on adipogenesis, preadipocyte differentiation was induced in the presence or absence of EGCG (0~100 μM) for a period of 6 days. EGCG significantly inhibited fat accumulation and suppressed the expression of adipogenic specific proteins including peroxisome proliferator-activated receptor (PPAR)-γ. Also, EGCG markedly increased the activation of AMPK and acetyl-CoA carboxylase (ACC) and the production of intracellular reactive oxygen species (ROS). However, any pretreatment with a specific AMPK inhibitor, compound C, abolished the inhibitory effects of the EGCG on PPARγ expression. This study suggests that EGCG has anti-adipogenic effects through modulation of the AMPK signaling pathway and therefore, may be a promising antiobesity agent.
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.
Background : The objective of this study was to Amomum tsao-ko Crevost et Lemarié, (Zingiberaceae) is widely distributed among several countries in South Asia and Southeast Asia. It is a well known spice in Asia, produces a nice refreshing effect in the mouth. Additionally, it's dried fruit is used in Traditional Chinese Medicine (TCM) for cardiac diseases, edema, eye trouble, skin, itch and impotence. The objective of this study was evaluated the inhibitory activity on adipogenesis in 3T3-L1 cells from A. tsao-ko. Methods and Results : The fruits of A. tsao-ko were extracted with 80% EtOH two times at room temperature. The EtOH extract was suspended in distilled water and partitioned with solvent to give CH2Cl2, EtOAc and n-BuOH. The CH2Cl2 was suspended in n-hexane and partitioned with solvent to give 50%, 70% and 90% MeOH. The purification of each fraction by column chromatography separation and HPLC analysis. Consequently, several constituents were isolated five known compounds. The identification and structural elucidation of compounds were established by using NMR (1D and 2D) and mass spectrometry. Conclusion : These compounds were identified as fluorenone (1), phenanthrene (2), anthracene (3), methyl linolenate (4), 1,2-benzenediol (5). All isolates were tested for their inhibitory activities on adipogenesis in 3T3-L1 cells.
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.
본 연구에서는 연화 열수 및 에탄올 추출물의 3T3-L1 지방세포 분화 및 ROS생성 저감효과를 구명하기 위하여 3T3-L1 전지방세포 분화과정 중 연화 열수 및 에탄올 추출 물에 의한 지방축적과 ROS 생성을 관찰하였다. 연화 열수 및 에탄올 추출물은 XTT assay에서 100, 200 및 400 μg/mL 농도에서 세포 독성을 보이지 않았다. 지방세포 분화 중 세포 내 지방축적 및 ROS 생성량을 비교한 결과, 연화 열수 및 에탄올 추출물을 처리한 지방세포의 경우 지방축적량과 ROS 생성량 모두 유의적으로 억제되는 것으로 나타났다. 특히 연화 열수 및 에탄올 추출물을 처리함으로써 지방세포 분화와 관련된 전사인자인 PPARγ, C/EBPα 및 aP mRNA의 발현을 유의적으로 감소시켰으며, ROS의 생성과 관련이 있는 주요 유전자인 NOX4 및 catalase의 유전자발현 또한 유의적으로 감소하였다. 이 결과를 통해 연화 열수 및 에탄 올 추출물이 3T3-L1 지방세포 내 중성지방의 축적 억제 효과와 더불어 ROS 생성 억제에 효과적으로 작용함을 확인 하였다. 따라서 연화 열수 및 에탄올 추출물은 비만과 같은 대사증후군 관련 질환의 개선을 위한 천연물 기능성 소재로 의 활용이 기대된다.
후박추출물은 중국 원산인 당후박에서 제조되며, 항산화 및 항염증 효과로 알려져 있다. 후박의 성분으로는 폴레페놀인 honokiol 및 magnolol 등이 알려져 있다. 이 연구에서, 우리는 후박추출물이 triglyceride (TG)의 축척을 감소시킴으로써 지방세포 분화를 억제한다는 표면적인 결과를 얻었다. 또한, 후박 추출물이 hormone sensitive lipase (HSL) 단백질 수준을 증가시키고 adipocyte에 specific한 transcription factors로 peroxisome proliferator activated receptor (PPAR)-γ의 단백질과 mRNA 수준을 억제하였다. 결과적으로, 후박추출물이 항비만제로서 adipogenic transcription factor와 그들의 특이적인 유전자의 발현을 억제함으로써 지방세포 분화를 억제한다고 사료된다.