• 생명과학회지
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• 제22권10호
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• pp.1316-1323
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• 2012

본 연구는 한우유래 myoblast에서 지방세포분화 유도 전사인자들을 과발현시켜 지방세포로의 교차분화를 유도하기 위하여 실시하였다. 한우 유래 satellite cell을 배양한 후 adipogenic transcription factor인 $PPAR{\gamma}$, C/$EBP{\alpha}$, SREBP1c, KLF5등을 단독 또는 co-transfection을 실시하여 세포에 과발현을 유도하였다. 이들 세포들은 adipogenic differentiation medium에서 2일간 배양한 후growth medium에서 8일간 추가로 배양하였다. 지방세포로의 교차분화 유무는 Oil-red O염색과 지방세포 마커 유전자들의 발현으로 확인하였다. $PPAR{\gamma}$과 C/$EBP{\alpha}$를 각각 단독으로 과발현을 유도한 경우myoblast에서 지방세포로의 교차분화를 유도하기에는 충분하지 못하였다. 그러나 $PPAR{\gamma}$와 C/$EBP{\alpha}$을 co-transfection을 실시한 경우 지방세포로의 교차분화가 유도되었고, 세포내지방구형성, 지방세포 마커유전자의 발현, 근세포 마커유전자의 발현 감소 등이 확인되었다. KLF5 와 $PPAR{\gamma}$를 동시에 과발현할 경우 지방세포로의 교차분화를 볼 수 있었지만 KLF단독의 경우는 교차분화를 유도하지 못하였다. 할성형SREBP1c (tSREBP1c)의 경우, 단독으로 myoblast에 과발현을 처리한 경우만으로 지방세포로의 교차분화를 유도할 수 있었다. 이들 결과는 한우유래 satellite cell을 이용하여 지방세포분화 전사인자를 단독 혹은 조합하여 이들 세포에 과발현 시킬 경우 지방세포로의 교차분화를 유도할 수 있음을 보여 주었다.

• Molecules and Cells
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• 제37권10호
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• pp.734-741
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• 2014

Histone modifications on major transcription factor target genes are one of the major regulatory mechanisms controlling adipogenesis. Plant homeodomain finger 2 (PHF2) is a Jumonji domain-containing protein and is known to demethylate the histone H3K9, a repressive gene marker. To better understand the function of PHF2 in adipocyte differentiation, we constructed stable PHF2 knock-down cells by using the mouse pre-adipocyte cell line 3T3-L1. When induced with adipogenic media, PHF2 knock-down cells showed reduced lipid accumulation compared to control cells. Differential expression using a cDNA microarray revealed significant reduction of metabolic pathway genes in the PHF2 knock-down cell line after differentiation. The reduced expression of major transcription factors and adipokines was confirmed with reverse transcription- quantitative polymerase chain reaction and Western blotting. We further performed co-immunoprecipitation analysis of PHF2 with four major adipogenic transcription factors, and we found that CCATT/enhancer binding protein (C/EBP)${\alpha}$ and C/EBP${\delta}$ physically interact with PHF2. In addition, PHF2 binding to target gene promoters was confirmed with a chromatin immunoprecipitation experiment. Finally, histone H3K9 methylation markers on the PHF2-binding sequences were increased in PHF2 knock-down cells after differentiation. Together, these results demonstrate that PHF2 histone demethylase controls adipogenic gene expression during differentiation.

• 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.

• 생명과학회지
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• 제27권12호
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• pp.1486-1493
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• 2017

된장은 콩 발효식품으로 주원료인 콩류에 Bacillus subtilis, Rizopus, Mucor와 Aspergillus species를 접종하여 발효시킨 메주를 소금물과 혼합하여 숙성 시킨 한국의 전통적인 발효식품이다. 본 연구에서는 동물실험을 통하여 검정콩 된장의 항비만 효과를 확인하였다. 항비만 효과의 확인은 혈중 TG, TC, 아디포넥틴과 렙틴의 레벨을 측정함과 동시에 지방합성에 관여하는 전사인자인 SREBP-1c과 PPAR-g의 mRNA와 단백질 발현 정도를 측정하였다. 고지방 식이에 검정콩 된장을 첨가한 그룹에서는 고지방식이로 인해 증가된 체중을 유의적으로 감소시킴을 확인하였다. 혈중 중성지방, 콜레스테롤과 렙틴의 레벨은 고지방식이를 섭취한 마우스에 비하여 검정콩 된장을 섭취한 마우스에서 감소하였으며 아디포넥틴의 분비량은 유의적으로 증가하였다. 이러한 결과가 지방 생성의 억제로부터 유도되는지를 조사하기 위하여 지방 합성에 관여하는 전사인자인 SREBP-1c과PPAR-g의 mRNA양과 단백질 발현을 측정한 결과 검정콩 된장을 섭취한 마우스에서 현저하게 감소하는 것을 확인하였다. 이러한 결과는 검정 콩 된장의 섭취가 지방대사와 지방 전사 인자의 활성을 감소시킨다는 것을 확인함으로써 검정콩 된장이 비만의 예방과 진행을 개선시킬 수 있음을 증명하였다.

• 동의생리병리학회지
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• 제28권3호
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• pp.288-295
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• 2014

Bojungchiseub-tang (BJCST) has been used in symptoms and signs of edema, dampness-phlegm, kidney failure, and so on. BJCST is also expected to have strong anti-obesity activities. However, little is known about the mechanisms of its inhibitory effects on adipocyte differentiation and adipogenesis. In the present study, we examined the effects and mechanism of BJCST on transcription factors and adipogenic genes of 3T3-L1 preadipocytes to understand its inhibitory effects on adipocyte differentiation and adipogenesis. Our results showed that BJCST significantly inhibited differentiation and adipogenesis of 3T3-L1 preadipocytes in a dose-dependent manner. To elucidate the mechanism of the effects of BJCST on lowering lipid content in 3T3-L1 adipocytes, we examined whether BJCST modulate the expressions of transcription factors to induce adipogenesis and adipogenic genes related to regulate accumulation of lipids. As a result, the expression of steroid regulatory element-binding protein (SREBP)1, cytidine-cytidine-adenosine-adenosine-thymidine (CCAAT)/enhancer binding proteins ${\alpha}$ ($C/EBP{\alpha}$), $C/EBP{\beta}$, $C/EBP{\delta}$, and peroxisome proliferator-activated receptor ${\gamma}$ ($PPAR{\gamma}$) genes, which induce the adipose differentiation, liver X receptor $(LXR){\alpha}$ and fatty acid synthase (FAS) genes, which induce lipogenesis and adipose-specific aP2, Adipsin, lipoprotein lipase (LPL), CD36, TGF-${\beta}$, leptin and adiponectin genes, which compose fat formation were decreased. BJCST also reduced the expression of acyl CoA oxidase (ACO) and uncoupling protein (UCP) genes related to lipid oxidation. In conclusion, BJCST could regulate transcript factor related to induction of adipose differentiation and inhibited the accumulation of lipids and expression of adipogenic genes.

• 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.

• 생명과학회지
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• 제29권2호
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• pp.209-214
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• 2019

본 연구에서는 괭생이모자반 에탄올 추출물의 항비만 효과에 대해 알아보고자 3T3-L1 preadipocyte에서 Oil Red O staining과 triglyceride content, adipogenesis의 발현에 대해 조사하였다. MTT assay를 통해 괭생이모자반 에탄올 추출물의 세포 독성을 확인 해 본 결과 $200{\sim}1,000{\mu}g/ml$ 농도에서 90% 이상의 세포 생존율을 확인 할 수 있었다. 또한 insulin, dexamethasone과 IBMX 복합물을 처리하여 분화를 유도하였을 때는 분화유도제를 처리하지 않은 군에 비해 활발하게 지방구를 형성하였으나, 괭생이모자반 에탄올 추출물을 250, 500, $1,000{\mu}g/ml$ 농도로 처리하였을 때는 농도 의존적으로 지방구 형성을 억제하였다. 이를 정량하여 triglyceride content (%)로 나타낸 결과 농도별로 13%, 16%, 23% 억제 효과를 나타내었다. 또한 Western blot assay를 통해 adipogenic transcription factor의 발현양상을 비교해 본 결과, 분화유도제 처리에 의해 $C/EBP{\alpha}$, $C/EBP{\beta}$와 $PPAR{\gamma}$의 발현이 증가하였으나, 괭생이모자반 에탄올 추출물을 처리하였을 때는 이러한 전사인자들의 발현이 농도 의존적으로 감소됨을 확인 할 수 있었다. 이상의 결과를 통해 괭생이모자반 에탄올 추출물은 adipogenic transcription factor의 발현을 조절하여 지방세포의 분화를 억제하였으며, 지방구와 triglyceride 생성을 감소시켜 최종적으로 비만억제 효과를 나타낸다는 것을 알 수 있었다.

• 대한화장품학회지
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• 제35권2호
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• pp.117-123
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• 2009

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)-${\gamma}$의 단백질과 mRNA 수준을 억제하였다. 결과적으로, 후박추출물이 항비만제로서 adipogenic transcription factor와 그들의 특이적인 유전자의 발현을 억제함으로써 지방세포 분화를 억제한다고 사료된다.

• 대한한방내과학회지
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• 제38권1호
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• pp.1-9
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• 2017

Objective: This study investigated the effect of purified Carthamus tinctorius (C. tinctorius) extracted with a hot water and ethanol method on adipogenic differentiation of mouse bone marrow-derived mesenchymal stromal stem cells (mBMSCs). Methods: The C. tinctorius was extracted using hot water and ethanol. The samples were concentrated by a rotary evaporator and were then dried using a freeze-dryer. The mBMSCs were cultured and maintained in a minimum essential medium eagle alpha (${\alpha}-MEM$) supplemented with 10% FBS and 1% antibiotic antimycotic solution. To induce adipogenic differentiation, the cells were treated with Dulbecco's modified eagle's medium-low glucose (DMEM-LG) containing 1 mg/mL insulin, 1 mM dexamethasone, and 0.5 mM 3-isobutyl-1-methylxanthine. To evaluate the adipogenic differentiation ability, oil-red O staining was performed after adipogenic differentiation for 21 days. The mRNA expression and protein level of adipogenic-related genes were quantified by quantitative real-time PCR and western blotting, respectively. Results: In the results of the MTT assay, no concentrations of C. tinctorius extracts showed toxicity on mBMSCs, so we fixed the treatment concentration of the extract at 100 ng/mL. In oil-red O staining, the water-C. tinctorius extract treatment significantly decreased adipogenic differentiation compared with the control and ethanol extract groups. The water-C. tinctorius extract group in particular showed reduced mRNA and protein expression of Peroxisome proliferator-activated receptor gamma ($Ppar{\gamma}$) and CCAAT/enhancer-binding protein alpha ($C/ebp{\alpha}$), which are adipogenic-related transcription factors. Conclusion: These data suggest that extract of C. tinctorius decreased the adipogenic differentiation of mBMSCs, while only water-C. tinctorius extract had an effect on different adipogenesis in mBMSCs. The C. tinctorius will be a useful therapeutic reagent for the prevention of obesity-related diseases such as diabetes, hyperlipidemia, coronary artery disease, and osteoporosis.

• 한방재활의학과학회지
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• 제24권1호
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• pp.1-12
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• 2014

Objectives This study was performed to evaluate the effects of fermented lotus extracts on prediabetes and hyperlipidemia in high fructose diet rats. Methods Extracts of lotus leaf and lotus root were fermented using 4 different probiotics separately, including Lactobacillus plantarum, Lactobacillus rhamnosus, Bifidobacterium breve, and Bifidobacterium longum. Expressions of adipogenic transcription factors including Adiponectin, GLUT-4, Leptin, PPAR gamma, Resistin and Visfatin were analyzed by Real time PCR and Western blotting analysis. Results Fermented lotus extracts reduced blood glucose. Fermented lotus extracts inhibited adipogenic transcription factors by inhibiting preadipocytes differentiation. The level of gene expression of Adiponectin, GLUT-4, Leptin, PPAR gamma, Resistin and Visfatin in relation to that of GAPDH were increase or decrease significantly with the Fermented lotus formulation group. Conclusions Fermented lotus extracts showed hypoglycemic and hypolipidemic effects by inhibiting preadipocyte differentiation and controlling insulin sensitivity in high fructose diet rats.