• 한방재활의학과학회지
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• 제24권4호
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• pp.1-13
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• 2014

Objectives This study was to investigate the antioxidative capacity, antiobesity effect and anti-diabetes effects of Mori Fructus and dried Mori Fructus in Raw 264.7 cells and 3T3-L1 cells. Methods 3 different types of Mori Fructus extracts (water 100%, ethanol 30%, ethanol 100%) were used in this study. And 3 different types of dried Mori Fructus extracts (water 100%, ethanol 30%, ethanol 100%) were used in this study. Total polyphenol compund, total favonoid compound, DPPH radical scavenging, ROS activity, NO, cell proliferation were measured in the experiment. Expressions of adipogenic transcription factors including $C/EBP-{\alpha}$, $C/EBP-{\beta}$, $PPAR-{\alpha}$, $PPAR-{\gamma}$, $AMPK-{\alpha}$ were analyzed by Real time PCR. Results Mori Fructus extracts measurements are higher than dried Mori Fructus extracts measurements at Total flavonoid compound and total flavonoid compound. Mori Fructus extracts measurements are lower than dried Mori Fructus extracts measurements at DPPH radical scavenging, ROS activity, NO. In RT-PCR analysis, there is a tendency that dried Mori Fructus extracts inhibit the expression of $C/EBP-{\alpha}$, $C/EBP-{\beta}$ genes. In RT-PCR analysis, there is a tendency that dried Mori Fructus extracts promote the expression of $PPAR-{\alpha}$, $PPAR-{\gamma}$, $AMPK-{\alpha}$ genes. Conclusions Mori Fructus is effective on inhibiting the oxidation and dried Mori Fructus is effective on inhibiting the obesity and diabetes.

• 대한본초학회지
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• 제29권2호
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• pp.15-21
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• 2014

Objectives : The purpose of this study was to investigate inhibitory effects of steamed Polygonatum odoratum extract (POE) on differentiation and adipogenesis in 3T3-L1 adipocytes. Methods : Polygonatum odoratum (P. odoratum) extract was extracted with ethyl acetate. Total phenolic and flavonoid contents in POE were measured for antioxidant activity. The spectrophotometric method was used to determine the DPPH and ABTS radical scavenging activity and ferric-reducing antioxidant potential (FRAP). MTT assay was examined for cell toxicity, oil red O staining was performed for intracelluar adipogenesis in differentiated 3T3-L1 adipocytes. Western blot analysis for measurement of CCAAT/enhancer-binding protein ${\alpha}$ ($C/EBP{\alpha}$), peroxisome proliferator-activated receptor${\gamma}$ ($PPAR{\gamma}$) and AMP-activated protein kinase (AMPK) expressions were performed. Results : The results revealed that POE has antioxidant activities. Contents of total polyphenolics and flavonoids were $50.83{\pm}1.52$ GAE mg/100g dry weight of POE and $17.05{\pm}2.47$ RE mg/100g dry weight of POE, respectively. DPPH radical scavenging activity, and FRAP in 10 mg/ml concentration were $92.1{\pm}0.6%$, $244.8{\pm}9.0{\mu}M$ Fe(II) and ABTS inhibition in 5 mg/ml concentration was $84.8{\pm}4.1%$. Treatment of POE in adipocytes inhibited the differentiation and adipogenesis of 3T3-L1 adipocytes compared to those of vehicle control. Additionally, protein expressions of $C/EBP{\alpha}$ and $PPAR{\gamma}$, major transcription factor for the adipogenic genes, were significantly decreased compared to those of vehicle control (p<0.05). Futhermore, phosphorylation of AMPK was increased in 3T3-L1 adipocytes treated with POE compared to that of vehicle control (p<0.05). Conclusions : we demonstrate that steamed P. odoratum extract (POE) has potentiating antioxidant activities, inhibits differentiation and lipid accumulation and also induces energy expenditure in adipocytes, which may contribute to antiobesity property.

• Journal of Microbiology and Biotechnology
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• 제18권1호
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• pp.80-87
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• 2008

The 3T3-L1 cell line is a well-established and commonly used in vitro model to assess adipocyte differentiation. Over the course of several days, confluent 3T3-L1 cells can be converted to adipocytes in the presence of an adipogenic cocktail. In this study, the effects of chitosan oligosaccharides (CO) on adipocyte differentiation of 3T3-L1 cells were studied. The CO significantly decreased lipid accumulation, a marker of adipogenesis, in a dose-dependent manner. The low molecular mass CO (1-3 kDa) were the most effective at inhibiting adipocyte differentiation. Moreover, mRNA expression levels of both CCAAT/enhancer-binding protein (C/EBP) ${\alpha}$ and peroxisome proliferator-activated receptor (PPAR) ${\gamma}$, the key adipogenic transcription factors, were markedly decreased by CO treatments. CO also significantly down regulated adipogenic marker proteins such as leptin, adiponectin, and resistin. Our results suggest a role for CO as antiobesity agents by inhibiting adipocyte differentiation mediated through the down regulated expression of adipogenic transcription factors and other specific genes.

• 생약학회지
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• 제42권2호
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• pp.201-208
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• 2011

Obesity occur from the imbalance between energy intake and energy expenditure. Obesity is a complex chronic disease that is suggested to cause other metabolic disorders such as type 2 diabetes, hyperlipidemia, hypertension, and arteriosclerosis. In this study, our purpose is to investigate the anti-hyperglycemic and anti-obesitic effects of Maydis stigma water extract in 3T3-L1 adipocytes and db/db mice. Maydis stigma water extract at dose of 100 and 500 ${\mu}g/ml$ slowly inhibited cell viability as compared to that of control in mature adipocytes. Also, the additions of 50 and 250 ${\mu}g/ml$ of Maydis stigma water extract significantly inhibited the lipid accumulations and CCAAT/enhancer-binding protein(C/EBP) ${\alpha}$ and peroxisome proliferator-activated receptor(PPAR) ${\gamma}$ expressions with dose-dependent manner in 3T3-L1 adipocytes. Maydis stigma water extract at 250, 500, and 1000 ${\mu}g/ml$ only showed the increasing pattern on lipolysis activity. The oral treatment of Maydis stigma water extract (100 or 400 mg/kg body weight) in db/db mice only showed tendency to decrease body weight, food efficiency ratio (FER), HbA1c, blood glucose, total cholesterol, triglyceride, and the adipocyte size of in db/db mice. However, Maydis stigma water extract increased the insulin level in a dose dependent manner. Thus these results indicate that Maydis stigma water extracxt inhibits adipogenesis through regulation of C/EBP${\alpha}$ and PPAR${\gamma}$ expressions in 3T3-L1 adipocytes and shows anti-hyperglycemic effect through increase of insulin secretion in db/db mice.

• 한방재활의학과학회지
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• 제28권3호
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• pp.13-25
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• 2018

Objectives We investigated anti-obesity effects of essential oils extracted from Cnidii Rhizoma (CR) in immature adipocytes to magnify it's clinical therapeutic usage. Methods Essential oil of CR was extracted with ethyl acetate or petroleum ether and through steam distillation, respectively. Oil red-O staining for monitoring its inhibition effect on adipogenesis and differentiation in murine 3T3-L1 adipocytes and 3-(4,5-methylthiazol-2-yl)-2,5-diphenyletetra zolium bromide (MTT) assay for cell safety were done. Also phospho-adenosine monophosphate (AMP)-activted protein kinase (P-AMPK), AMP-activated protein kinase, phospho-acetyl-CoA carboxylase (P-ACC), acetyl-CoA carboxylase, peroxisome proliferator-activated receptor-${\alpha}$ (PPAR-${\alpha}$), peroxisome proliferator-activated receptor-${\gamma}$ (PPAR-${\gamma}$) and CCAAT/enhancer binding protein ${\alpha}$ (C/EBP-${\alpha}$) expressions as obesity-related factors were measured by western blot analysis. Results Protein expressions of P-AMPK, P-ACC and PPAR-${\alpha}$ were increased in essential oils-treated adipocytes compared to those of control group, respectively. Furthermore, protein expressions of PPAR-${\gamma}$ and C/EBP-${\alpha}$ were decreased in essential oils-treated adipocytes compared to those of control group, respectively. Conclusions These results demonstrate that essential oils of CR inhibit adipogenesis and differentiation. Also they promote the oxidation of fatty acids in adipocytes. Thus, results suggest that essential oils of CR could be used as a valuable material for anti-obesity therapeutics via control of lipid metabolism.

• 생명과학회지
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• 제24권5호
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• pp.476-484
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• 2014

비만은 감염원 없이 진행되는 낮은 수준의 만성적인 전신성 염증상태로 간주되며, 만성대사질환의 원인이 된다. 본 연구에서는 미나리를 발효하여 만든 식초가 3T3-L1 지방전구세포의 분화 및 RAW 264.7 세포에서의 항염증 효과에 미치는 영향을 확인하였다. MDI (IBMX, dexamethasone, insulin)에 의해 분화가 유발된 3T3-L1 지방전구세포에 대한 미나리 식초의 분화 억제능을 Oil-red-O staining을 통해 확인하였으며, western blot법을 통해 지방세포 형성 시 중요한 전사인자인 peroxisome proliferator-activated receptor-${\gamma}$ (PPAR-${\gamma}$)와 CCAAT-enhancer-binding protein ${\alpha}$ ($C/EBP{\alpha}$)의 발현을 감소시키는 것으로 나타났다. 또한, 미나리 식초가 lipopolysaccharide (LPS)에 의해 염증이 유발 된 RAW 264.7 세포의 nitric oxide (NO) 생성을 억제시켰으며, inducible NO synthase (iNOS)와 cyclooxygenase-2 (COX-2)의 단백질 발현을 감소시키는 것을 확인하였다. 이러한 결과들을 통해 미나리 발효 식초는 지방세포 분화와 염증 억제 효과를 가지고 있음을 확인하였다. 따라서 미나리 발효 식초는 대사성 질환을 예방할 수 있는 천연물 소재로 이용 가능할 것으로 생각된다.

• 한국식품영양과학회지
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• 제45권8호
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• pp.1107-1113
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• 2016

지방전구세포가 지방세포로 분화하는 과정에서 아선약 추출물의 영향을 확인하고자 분화유도제를 처리하여 3T3-L1 세포에서 아선약의 세포독성, 지방축적 억제 효과, triglyceride 억제 효능, 지방분화 관련 단백질들의 발현을 확인하였다. 아선약 추출물을 최대 $200{\mu}g/mL$ 농도까지 세포에 처리하여 세포독성을 측정한 결과 아선약 추출물은 $100{\mu}g/mL$ 이하의 농도에서는 세포에 독성을 유발하지 않는 것으로 확인되었다. Oil red-O 염색을 통해 지방축적률을 확인한 결과 아선약 추출물에 의한 농도 의존적인 지방축적의 감소 효과가 있었다. Triglyceride 생성량 역시 농도 의존적인 감소가 확인되었다. 아선약에 의한 지방분화의 감소가 어떠한 기작에 의해 조절되는지 확인하고자 관련 지표단백질인 $PPAR{\gamma}$와 SREBF-1 그리고 $C/EBP{\alpha}$의 발현량 변화를 확인해본 결과 $100{\mu}g/mL$ 농도에서 유의성 있게 감소하였다. 이상의 결과를 종합해볼 때 아선약 추출물은 지방세포의 분화억제와 분화된 지방세포의 지방축적을 저해함으로써 항비만 효과를 유도할 것으로 기대된다.

• Nutrition Research and Practice
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• 제12권6호
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• pp.494-502
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• 2018

BACKGROUND/OBJECTIVES: Reducing the number of adipocytes by inducing apoptosis of mature adipocytes as well as suppressing differentiation of preadipocytes plays an important role in preventing obesity. This study examines the anti-adipogenic and pro-apoptotic effect of red pepper seed water extract (RPS) prepared at $4^{\circ}C$ (RPS4) in 3T3-L1 cells. MATERIALS/METHODS: Effect of RPS4 or its fractions on lipid accumulation was determined in 3T3-L1 cells using oil red O (ORO) staining. The expressions of AMP-activated protein kinase (AMPK) and adipogenic associated proteins [peroxisome proliferator-activated receptor-${\gamma}$ ($PPAR-{\gamma}$), CCAAT/enhancer-binding proteins ${\alpha}$ (C/EBP ${\alpha}$), sterol regulatory element binding protein-1c (SREBP-1c), fatty acid synthase (FAS), and acetyl-CoA carboxylase (ACC)] were measured in 3T3-L1 cells treated with RPS4. Apoptosis and the expression of Akt and Bcl-2 family proteins [B-cell lymphoma 2 (Bcl-2), Bcl-2-associated death promoter (Bad), Bcl-2 like protein 4 (Bax), Bal-2 homologous antagonist/killer (Bak)] were measured in mature 3T3-L1 cells treated with RPS4. RESULTS: Treatment of RPS4 ($0-75{\mu}g/mL$) or its fractions ($0-50{\mu}g/mL$) for 24 h did not have an apparent cytotoxicity on pre and mature 3T3-L1 cells. RPS4 significantly suppressed differentiation and cellular lipid accumulation by increasing the phosphorylation of AMPK and reducing the expression of $PPAR-{\gamma}$, C/EBP ${\alpha}$, SREBP-1c, FAS, and ACC. In addition, all fractions except ethyl acetate fraction significantly suppressed cellular lipid accumulation. RPS4 induced the apoptosis of mature adipocytes by hypophosphorylating Akt, increasing the expression of the pro-apoptotic proteins, Bak, Bax, and Bad, and reducing the expression of the anti-apoptotic proteins, Bcl-2 and p-Bad. CONCLUSIONS: These finding suggest that RPS4 can reduce the numbers as well as the size of adipocytes and might useful for preventing and treating obesity.

• 대한본초학회지
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• 제33권2호
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• pp.69-77
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• 2018

Objectives : Daesiho-tang (DSHT) has been widely used in the treatment of cerebral infarct in traditional medicine. However, there was not report on the anti-obesity-related diseases efficacy of DSHT. In this study, we investigated the effects for the new formulation of DSHT, on the adipocyte differentiation cycle in 3T3-L1 cells. Methods : 3T3-L1 cells were treated with DSHT (50, 100, $200{\mu}g/m{\ell}$) during differentiation for 6 days. Also, the inhibitory effect of DSHT against 3T3-L1 adipogenesis was evaluated in various stage of adipogenesis such as early (0-2day), intermediate (2-4day), and terminal stage (4-6day). The accumulation of lipid droplets was determined by Oil Red O staining. and, the expressions of genes related to adipogenesis were measured by RT-PCR and Western blot analyses. Results : DSHT showed inhibitory activity on adipocyte differentiation at 3T3-L1 preadipocytes without affect cell toxicity as assessed by measuring fat accumulation and adipogenesis. In addition, DSHT significantly reduced the expression levels of several adipocyte marker genes including proliferator activated $receptor-{\gamma}$ ($PPAR-{\gamma}$) and CCAAT/ enhancer-binding $protein-{\alpha}$ ($C/EBP-{\alpha}$). Also, the anti-adipogenic effect of DSHT was strongly limited in the intermediate (2-4 day), terminal stage (4-6 day) of 3T3-L1 adipogenesis. In addition, the DSHT treatment down- regulated mRNA expression levels of $PPAR-{\gamma}$,, $C/EBP-{\alpha}$ in mature 3T3-L1 adipocytes. Conclusions : These results suggest that, the ability of DSHT has inhibited overall adipogenesis and lipid accumulation in the 3T3-L1 cells. The new formulation of DSHT may be a promising medicine for the treatment of obesity and related metabolic disorders.

• 생약학회지
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• 제41권4호
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• pp.270-274
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• 2010

Obesity is caused from an imbalance between energy intake and expenditure, which may lead to pathologic growth of adipocytes and accumulation of fat in tissue. We examined the inhibitory effects of Eriobotrya japonica leaf and seed extracts on lipid absorption in vitro and fat accumulation during the differentiation of 3T3-L1 to adipocytes. 3T3-L1 preadipocytes were stimulated with DMEM media containing 10% FBS, 0.5 mM 3-isobuthyl-1-methyxanthine (IBMX), $5\;{\mu}g/ml$ insulin, and $1\;{\mu}g/ml$ dexamethasone for differentiation to adipocytes. E. japonica leaf extract at concentration of 0.5 or 1 mg/ml inhibited pancreatic lipase activity. The cell viability of 3T3-L1 adipocytes slightly reduced about 3% by treatment of E. Japonica leaf and seed extracts. The leaf and seed extracts of E. japonica effectively inhibited the accumulations of lipid droplet and expression of $C/EBP{\alpha}$ promoting adipogenesis. Thus, this data suggest that E. japonica leaf and seed extracts inhibit fat accumulation through regulation of $C/EBP{\alpha}$, and leaf extract is more effective in lipid absorption and adipogenesis than seed extract.