• 한국식품영양과학회지
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• 제41권7호
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• pp.895-900
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• 2012

본 연구에서 동과 물추출물의 계통분획을 통해 획득된 세가지 분획물인 핵산 분획(BHHH), 클로로포름 분획(BHHC), 에틸아세테이트 분획(BHHE)들을 3T3-L1 분화과정 중에 처리한 후, Oil Red O 염색법에 의한 lipid accumulation, 지방구내 triglyceride 함량을 평가하고, free glycerol release 함량과 adipogenesis와 관련된 transcription factor들의 발현 함량을 비교하여 동과 물추출물 중 anti-adipogenesis 활성 분획물을 밝히고, 이 분획물의 작용 메커니즘을 규명하고자 하였다. 50 ${\mu}g/mL$ 농도의 BHHC와 BHHE의 처리는 분화된 지방세포 내 지질 축척을 11%와 13%로 낮추었다. 지방세포 내 중성지방(TG)의 함량은 동일 농도의 각 분획물에서 21%와 16%로 낮게 나타났다. TG 함량의 감소와 지방구내 지질 축적의 감소, 즉 anti-adipogenesis 메커니즘을 밝히기 위해 free glycerol 분비량을 평가하였다. 동일 농도의 BHHC와 BHHE에서 각각 13%와 17% 감소되어 나타났다. BHHC와 BHHE는 세포가 분화하는 동안 $PPAR{\gamma}$, C/$EBP{\alpha}$, leptin의 mRNA 발현을 억제하는 것으로 나타났다. 특히 BHHE의 경우 각 transcriptional factor들의 발현을 45%, 67%, 35%로 현저히 억제시키는 우수한 anti-adipogenetic 소재로 나타났다. 이에 BHHE는 항비만 기능성 소재로 활용될 수 있을 것으로 판단된다.

• 생명과학회지
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• 제24권10호
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• pp.1085-1091
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• 2014

본 연구는 지방전구세포에서 selenate가 지방세포의 분화와 초기 분화조절 연관 selenoprotein의 유전자발현 양상을 조사하기 위하여 실시하였다. Selenate의 농도를 달리하여($0-100{\mu}M$) 지방전구세포가 confluent한 시기, 지방세포분화 유도 초기(day0-day2), 분화유도 중기(day2-day4), 또는 분화전기간(day0-day6)으로 구분하여 처리한 다음 세포내 지방구형성과 소포체(ER)스트레스 및 selenoptotein 유전자들의 발현 양상을 분석하였다. Selenate에 의한 지방세포분화 억제효과는 지방세포가 post-confluent (cell cycle arrest), 분화유도 초기(mitotic clonal expansion), 또는 지속적인 처리가 주어진 상태에서 농도 의존적으로 나타났다(p<0.05). 그러나 selenate를 분화유도 중기(post-mitotic growth arrest)에 처리되었을 때는 세포분화억제 효과가 감소하였다. Seps1와 Sepp1의 selenoprotein 유전자들은 mitotic clonal expansion시기에 높게 발현하였다가 post-mitic growth period에는 급격한 발현감소현상을 나타내었다(p<0.05). 본 연구 결과는 selenate에 의한 지방전구세포의 분화억제효과는 세포분화초기에 효과적인 기능이 있음을 보여 주었으며, selenoprotein의 분화유도초기와 중기의 변환지점에 이들 유전자들의 급격한 발현변화가 selenate에 의한 지방세포의 분화억제와 관련이 있는 것으로 보여진다.

• 대한본초학회지
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• 제31권4호
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• pp.71-77
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• 2016

Objectives : Adipogenesis was defined as a differentiation process of preadipocytes into the adipocytes. Thus, to control of this process can be one of the most important strategies to prevent obesity. Korean ginseng(Panax ginseng C.A. Meyer) is one of the most widely used medicinal herbs. Although multiple biological activities of Korean ginseng, particularly ginsenosides, have been known, the anti-adipogenic role and function of polysaccharides from Korean ginseng are still unclear. In this study, we examined anti-adipogenic activity of polysaccharides and its molecular basis mechanisms are further investigated.Methods : The cytotoxicity of KGP in 3T3-L1 was evaluated by MTT assay. Anti-adipogenic effect of KGP was examined by Oil Red O (ORO) staining and microscopy observation in 3T3-L1 mature adipocytes. The mRNA expression levels of adipogenic transcriptional factors were analyzed by reverse transcription-polymer chain reaction (RT-PCR). To elucidate the adipogenic molecular mechanism of KGT, SB431542 (TGF-β specific inhibitor) was used.Results : We found that polysaccharides showed no effect on the viability of 3T3-L1 preadipocytes. Dose dependent inhibitory effect of polysaccharides on 3T3-L1 adipogenesis was observed as judged by ORO staining and microscopic image analysis. To obtain further mechanistic insight into anti-adipogenic function of polysaccharides, we then tested the effect of polysaccharides treatment on the adipogenic marker genes. The mRNA expressions level of C/EBPα, PPARγ, C/EBPβ, and fatty acid synthase (FAS) were dose-dependently inhibited by KGP treatment in 3T3-L1 mature adipocytes.Conclusions : In conclusion, these findings suggest that the KGP could be used in treatment of obesity and overweight related diseases.

• Preventive Nutrition and Food Science
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• 제20권4호
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• pp.298-302
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• 2015

The anti-obesity effects of starter (Leuconostoc mesenteroides+Lactobacillus plantarum) fermented kimchi on 3T3-L1 adipocyte were studied using naturally fermented kimchi (NK), a functional kimchi (FK, NK supplemented with green tea), and FK supplemented with added starters (FKS). Oil red O staining and cellular levels of triglyceride (TG) and glycerol were used to evaluate the in vitro anti-obesity effects of these kimchis in 3T3-L1 cells. The expressions of adipogenesis/lipogenesis-related genes of peroxisome proliferator-active receptor (PPAR)-${\gamma}$, CCAAT/enhance-binding protein (C/EBP)-${\alpha}$, and fatty acid synthase (FAS) were determined by RT-PCR. Kimchis, especially FKS, markedly decreased TG levels and increased levels of intracellular glycerol and lipid lipolysis. In addition, FKS also reduced the mRNA levels of PPAR-${\gamma}$, C/EBP-${\alpha}$, and FAS, which are related to adipogenesis/lipogenesis in 3T3-L1 cells. These results suggest the anti-obesity effects of FKS were to due to enhanced lipolysis and reduced adipogenesis/lipogenesis in 3T3-L1 adipocytes.

• 대한의용생체공학회:의공학회지
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• 제41권3호
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• pp.138-145
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• 2020

Adipocytes are the main constituent of adipose tissue. Understanding the molecular basis of adipogenesis is pivotal to finding the therapeutic targets for treatment of obesity. Melatonin is associated with obesity and its mechanism is currently under intensive investigation. The objective of this study was to investigate the effect of melatonin on adipogenesis in differentiating preadipocytes. 3T3-L1 preadipocytes were cultured in Dulbecco's modified Eagle's medium (DMEM) containing 5% calf serum at 37℃ with 5% CO₂ in a humidified incubator. Differentiation was induced using DMEM with 10% fetal bovine serum supplemented with MDI two days after cell confluence (day 0). Cells were treated with 0, 10 and 100 μM melatonin on either day 0 or day 5. 72 hours after each treatment, lipid accumulation was measured by oil red O staining. Proteins were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to membranes. As a result, lipid accumulation decreased with melatonin treatment. ERK pathway, activated when differentiation is induced, also decreased with an increase in melatonin concentration. Furthermore, the expression of key adipogenic factors, C/EBPα, C/EBPβ, and PPARγ, were reduced by melatonin treatment. These results imply that melatonin may inhibit the process of adipogenesis and may have a role as a new anti-obesity agent.

• Nutrition Research and Practice
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• 제18권2호
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• pp.180-193
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• 2024

BACKGROUND/OBJECTIVES: Obesity is a major cause of metabolic disorders; to prevent obesity, research is ongoing to develop natural and safe ingredients with few adverse effects. In this study, we determined the anti-obesity effects of Rosa multiflora root extract (KWFD-H01) in 3T3-L1 adipocytes and Sprague-Dawley (SD) rats. MATERIALS/METHODS: The anti-obesity effects of KWFD-H01in 3T3-L1 adipocytes and SD rats were examined using various assays, including Oil Red O staining, gene expression analyses, protein expression analyses, and blood biochemical analyses. RESULTS: KWFD-H01 reduced intracellular lipid accumulation and inhibited the mRNA expression of peroxisome proliferator-activated receptor γ (PPARγ), cytidine-cytidine-adenosine-adenosine-thymidine (CCAAT)/enhancer binding proteins (C/EBPα), sterol regulatory element-binding transcription factor 1 (SREBP-1c), acetyl-CoA carboxylase (ACC), and fatty acid synthase (FAS) in 3T3-L1 cells. KWFD-H01 also reduced body weight, weight gain, and the levels of triglycerides, total and LDL-cholesterol, glucose, and leptin, while increasing high-density lipoprotein-cholesterol and adiponectin in SD rats. PPARγ, C/EBPα, SREBP-1c, ACC, and FAS protein expression was inhibited in the epididymal fat of SD rats. CONCLUSION: Overall, these results confirm the anti-obesity effects of KWFD-H01 in 3T3-L1 adipocytes and SD rats, indicating their potential as baseline data for developing functional health foods or pharmaceuticals to control obesity.

• 한국식품영양학회지
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• 제30권5호
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• pp.1035-1041
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• 2017

(-)-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 ($0{\sim}100{\mu}M$) 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)-${\gamma}$. 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 $PPAR{\gamma}$ 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.

• Nutrition Research and Practice
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• 제8권6호
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• pp.613-617
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• 2014

BACKGROUND/OBJECTIVES: Adipogenesis is part of the cell differentiation process in which undifferentiated fibroblasts (pre-adipocytes) become mature adipocytes with the accumulation of lipid droplets and subsequent cell morphological changes. Several transcription factors and food components have been suggested to be involved in adipogenesis. The aim of this study was to determine whether mulberry leaf ethanol extract (MLEE) affects adipogenesis in 3T3-L1 adipocytes. MATERIALS/METHODS: The 3T3-L1 adipocytes were treated with different doses of MLEE for 8 days starting 2 days post-confluence. Cell viability, fat accumulation, and adipogenesis-related factors including CCAAT-enhancer-binding protein alpha ($C/EBP{\alpha}$), peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$), $PPAR{\gamma}$ coactivator 1 alpha (PGC-$1{\alpha}$), fatty acid synthase (FAS), and adiponectin were analyzed. RESULTS: Results showed that MLEE treatments at 10, 25, 50, and $100{\mu}g/ml$ had no effect on cell morphology and viability. Without evident toxicity, all MLEE treated cells had lower fat accumulation compared with control as shown by lower absorbances of Oil Red O stain. MLEE at 50 and $100{\mu}g/ml$ significantly reduced protein levels of $PPAR{\gamma}$, PGC-$1{\alpha}$, FAS, and adiponectin in differentiated adipocytes. Furthermore, protein level of $C/EBP{\alpha}$ was significantly decreased by the treatment of $100{\mu}g/ml$ MLEE. CONCLUSION: These results demonstrate that MLEE treatment has an anti-adipogenic effect in differentiated adipocytes without toxicity, suggesting its potential as an anti-obesity therapeutic.

• 대한한방소아과학회지
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• 제24권3호
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• pp.92-105
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• 2010

Objectives: The purpose of this study is to investigate the effects of Daecheongryong-tang (DCRT) on the adipogenesis in 3T3-L1 preadipocytes. Methods: 3T3-L1 preadipocytes were differentiated with adipogenic reagents by incubating for 2 days in the absence or presence of DCRT ranging 0.25 and 2%. The effect of DCRT on adipogenesis was examined by Oil red O staining, and the protein, RNA, and RT-PCR were measured. Results: Our results showed that DCRT decreased the TG content by ORO staining. To elucidate the mechanism of the effects of DCRT on lowering TG content in 3T3-L1 adipocytes, we examined the DCRT modulate expressions of transcription factors to induce adipogenesis and adipogenic genes which is related to the regulation of accumulation of lipids. As a result, the expression of SREBP1, C/$EBP{\beta}$, C/$EBP{\delta}$, C/$EBP{\alpha}$, and $PPAR{\gamma}$ genes, which induce the adipose differentiation and adipose-specific aP2, adipsin, LPL, CD36, TGF-${\beta}$ and adiponectin genes which regulates fat formations, were decreased. In addition, DCRT reduced the expression of iNOS and IL-6 in 3T3-L1 adipocytes, resulting in inflammation. Conclusions: DCRT could regulate transcript factor related to induction of adipose differentiation, inhibit the accumulation of lipids and expression of the adipogenic genes.

• Nutrition Research and Practice
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• 제15권4호
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• pp.444-455
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• 2021

BACKGROUND/OBJECTIVES: Adipocytes undergo angiogenesis to receive nutrients and oxygen needed for adipocyte' growth and differentiation. No study relating quercetin with angiogenesis in adipocytes exists. Therefore, this study investigated the role of quercetin on adipogenesis in 3T3-L1 cells, acting through matrix metalloproteinases (MMPs). MATERIALS/METHODS: After proliferating preadipocytes into adipocytes, various quercetin concentrations were added to adipocytes, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays were performed to evaluate cell proliferation. Glycerol-3-phosphate dehydrogenase (GPDH) activity was investigated as an indicator of fat accumulation. The mRNA expressions of transcription factors related to adipocyte differentiation, CCAAT/enhancer-binding proteins (C/EBPs), peroxisomal proliferatoractivated receptors (PPAR)-γ, and adipocyte protein 2 (aP2), were investigated. The mRNA expressions of proteins related to angiogenesis, vascular endothelial growth factor (VEGF)-α, vascular endothelial growth factor receptor (VEGFR)-2, MMP-2, and MMP-9, were investigated. Enzyme activities and concentrations of MMP-2 and MMP-9 were also measured. RESULTS: Quercetin treatment suppressed fat accumulation and the expressions of adipocyte differentiation-related genes (C/EBPα, C/EBPβ, PPAR-γ, and aP2) in a concentration-dependent manner in 3T3-L1 cells. Quercetin treatments reduced the mRNA expressions of VEGF-α, VEGFR-2, MMP-2, and MMP-9 in 3T3-L1 cells. The activities and concentrations of MMP-2 and MMP-9 were also decreased significantly as the concentration of quercetin increased. CONCLUSIONS: The results confirm that quercetin inhibits adipose tissue differentiation and fat accumulation in 3T3-L1 cells, which could occur through inhibition of the angiogenesis process related to MMPs.