• KSBB Journal
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• 제27권6호
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• pp.381-386
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• 2012

Obesity increases the risk of many adult diseases, such as atherosclerosis, diabetes, hypertension, ischemic heart disease and breast cancer. Inhibition of adipogenesis is an effective way in the anti-obesity management. Because of main components of Saururus chinensis is flavonoid, it has been showed some improvement by its antioxidant effects on the atherosclerosis, heart disease and diabetic hyperlipidemia. But mechanism of anti-obesity effect of S. chinensis is not clear. We investigated the effects of ethanol extracts of S. chinensis on adipogenesis in the 3T3-L1 pre-adipocyte. The 3T3-L1 cell line is commonly used to study adipogenesis in vitro. In this study, ethanol extracts of S. chinensis significantly decrease the lipid accumulation in the 3T3-L1 cells proved by measuring triglyceride contents and Oil red O staining. The proposed mechanism of inhibition of adipogenesis in the 3T3-L1 cells with ethanol extracts of S. chinensis is down-regulation of transcriptional factors and adipocyte-specific genes such CCAAT/enhancer binding protein ${\alpha}$ ($C/EBP{\alpha}$) and Peroxisome proliferator activated receptor ${\gamma}$ ($PPAR{\gamma}$) in concentration dependent pattern. These results suggest that ethanol extracts of S. chinensis inhibits adipognesis in the 3T3-L1 cells and can be used as a safe and efficient natural substance to manage anti-obesity.

• 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.

• Molecules and Cells
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• 제38권4호
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• pp.336-342
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• 2015

Propyl gallate (PG) used as an additive in various foods has antioxidant and anti-inflammatory effects. Although the functional roles of PG in various cell types are well characterized, it is unknown whether PG has effect on stem cell differentiation. In this study, we demonstrated that PG could inhibit adipogenic differentiation in human adipose tissue-derived mesenchymal stem cells (hAMSCs) by decreasing the accumulation of intracellular lipid droplets. In addition, PG significantly reduced the expression of adipocyte-specific markers including peroxisome proliferator-activated receptor-${\gamma}$ (PPAR-${\gamma}$), CCAAT enhancer binding protein-${\alpha}$ (C/EBP-${\alpha}$), lipoprotein lipase (LPL), and adipocyte fatty acid-binding protein 2 (aP2). PG inhibited adipogenesis in hAMSCs through extracellular regulated kinase (ERK) pathway. Decreased adipogenesis following PG treatment was recovered in response to ERK blocking. Taken together, these results suggest a novel effect of PG on adipocyte differentiation in hAMSCs, supporting a negative role of ERK1/2 pathway in adipogenic differentiation.

• 한국수정란이식학회지
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• 제32권3호
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• pp.193-200
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• 2017

The purpose of this study is to confirm whether spontaneous adipocyte generation during chondrogenic induction culture affects the chondrogenic differentiation of porcine skin-derived stem cells (pSSCs). For this purpose, chondrogenic differentiation characteristics and specific marker gene expression were analyzed using cell lines showing different characteristics of spontaneous adipocyte formation. Of the four different lines of pSSCs, the pSSCs-IV line showed higher Oil red O (ORO) and glycosaminoglycan (GAG) extraction levels. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis revealed that the levels of adipogenic markers peroxisome proliferator-activated receptor gamma 2 ($PPAR{\gamma}2$) and adipocyte Protein 2 (aP2) mRNAs were significantly higher in pSSCs-IV than those of the other pSSC lines (P<0.05). Among three chondrogenic markers, collagen type II (Col II) and sex determining region Y-box (Sox9) mRNAs were strongly expressed in pSSCs-IV (P<0.05), but not in aggrecan (Agg), which was significantly higher in pSSCs-II (P<0.05). These results demonstrate that the spontaneous adipocyte generation during chondrogenic differentiation has a positive effect on the chondrogenesis of pSSCs. More research is needed on the correlation between adipocyte generation and cartilage formation.

• 한국약용작물학회지
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• 제25권6호
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• pp.367-374
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• 2017

Background: An imbalance in energy intake and expenditure can cause obesity, which is a major risk factor for chronic diseases such as heart disease, type 2 diabetes, insulin resistance, cancers and hyperlipidemia. Methods and Results: In this study, we evaluated the anti-obesity effects of a water extract from the young leaves of barley sprout (BS) in 3T3-L1 cells and in high-fat diet (HFD)-induced obese mice (HF). Lipid accumulation measurement indicates that BS markedly inhibited adipogenesis by reducing lipid droplet production in a dose-dependent manner. Furthermore, the mRNA expression of adipogenic transcription factors peroxisome proliferator-activated receptor-${\gamma}$ and fatty acid synthetase, CCAAT/enhancer binding protein-${\alpha}$ and fatty acid binding protein 4 in 3T3-L1 cells was significantly inhibited by BS treatment. In an in vivo test, the BS-administered group of HFD-induced mice showed less body weight gain, and lower liver and epididymal white adipose tissue weights. The BS-treated mice showed decreased serum levels of leptin and lipids compared to untreated HFD mice and the levels of adiponectin and the HDL-cholesterol/total cholesterol ratio increased. These results indicate that BS inhibits body fat accumulation by reducing the mRNA expression of lipogenesis transcription factors and increasing serum adipokine concentration in in vitro and in vivo tests. Conclusions: BS reduced high fat diet-induced weight gain and had a positive effect on dyslipidemia.

• 한국식품영양과학회지
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• 제44권3호
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• pp.324-330
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• 2015

본 연구에서는 C57BL/6J ob/ob 마우스를 이용하여 한국산 겨우살이 냉수 추출물(KME)의 항당뇨 활성을 조사하였다. 50 혹은 100 mg/kg의 KME를 1일 1회씩 경구투여 한 결과 KME 투여 개시 5일 후부터 ob/ob 마우스의 혈당이 유의하게 억제되었으며, 10일 후부터 안정된 억제 효과를 나타내고 대조군에 비해 20% 이상의 혈당강하 효과를 나타내었다. 경구 당부하 실험(OGTT)에서는 KME 경구투여 마우스에서 유효한 당부하 억제 활성이 관찰되었다. 또한 KME 경구 투여는 ob/ob 당뇨 마우스의 혈액 내 총 콜레스테롤과 중성 지질의 농도를 억제하는 것으로 나타났다. 한편 C2C12 근육세포를 이용한 in vitro 실험에서 KME를 처리함으로써 glucose uptake가 현저히 증가하였다. 한편 매우 흥미롭게도 KME를 처리한 C2C12 근육세포에 있어서 미토콘드리아 생성과 산화대사 조절물질인 peroxisome proliferator-activated receptor gamma coactivator 1-${\alpha}(PGC-1{\alpha})$를 비롯하여 glucose transporter type 4(GLUT4), estrogen-related receptor-${\alpha}(ERR-{\alpha})$, nuclear respiratory factor-1(NRF-1) 그리고 mitochondrial transcription factor A(TmfA)와 같은 $PGC-1{\alpha}$ 관련 유전자들의 발현이 증가하는 것으로 확인되었다. 이 결과는 KME가 2형 당뇨에 대한 치료물질로서의 작용을 지니며 이러한 KME의 항당뇨 활성은 미토콘드리아 생성의 조절과 관련 있는 것으로 추정된다.

• Journal of Microbiology and Biotechnology
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• 제18권6호
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• pp.1170-1178
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• 2008

The cyclopentenone prostaglandins (cyPGs) prostaglandin $A_1$ ($PGA_1$) and 15-deoxy-${\Delta}^{12,14}$-prostaglandin $J_2$ (15d-$PGJ_2$) have been reported to exhibit antiinflammatory activity in activated monocytes/macrophages. However, the effects of these two cyPGs on the expression of cytokine genes may differ. In this study, we investigated the mechanism of action of $PGA_1$ in lipopolysaccharide (LPS)-induced expression of inter leu kin (IL)-10 mRNA in mouse peritoneal macrophages. 15d-$PGJ_2$ inhibited expression of LPS-induced IL-10, whereas $PGA_1$ increased LPS-induced IL-10 expression. This synergistic effect of $PGA_1$ on LPS-induced IL-10 expression reached a maximum as early as 2 h after simultaneous $PGA_1$ and LPS treatment ($PGA_1$/LPS), and did not require new protein synthesis. The synergistic effect of $PGA_1$ was inhibited by GW9662, a specific peroxisome proliferator-activated receptor ${\gamma}(PPAR{\gamma})$ antagonist, and Bay-11-7082, a NF-${\kappa}B$ inhibitor. The extracellular signal-regulated kinases (ERK) inhibitor PD98059 increased the expression of $PGA_1$/LPS-induced IL-10 mRNA, rather than inhibiting the IL-10 expression. Moreover, $PGA_1$ inhibited LPS-induced ERK phosphorylation. The synergistic effect of $PGA_1$ on LPS-induced IL-10 mRNA and protein production was inhibited by p38 inhibitor PD169316, and $PGA_1$ increased LPS-induced p38 phosphorylation. In the case of stress-activated protein kinase/c-Jun $NH_2$-terminal kinase (SAPK/JNK), the SAPK/JNK inhibitor SP600125 did not inhibit IL-10 mRNA synthesis but inhibited the production of IL-10 protein remarkably. These results suggest that the synergistic effect of $PGA_1$ on LPS-induced IL-10 expression is NF-${\kappa}B$-dependent and mediated by mitogen-activated protein (MAP) kinases, p38, and SAPK/JNK signaling pathways, and also associated with the $PPAR{\gamma}$ pathway. Our data may provide more insight into the diverse mechanisms of $PGA_1$ effects on the expression of cytokine genes.

• 생명과학회지
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• 제21권9호
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• pp.1346-1353
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• 2011

산두근(Sophora tonkinensis Gapnep)은 예로부터 동양지역에서 전통적인 약용식물로 사용되어 왔다. 본 연구에서는 3T3-L1 지방전구세포의 성숙지방세포로의 분화와 세포 내 지방생성에 대한 산두근 메탄올 추출물(STME)의 효과와 메커니즘에 대해 조사하였다. STME를 0-200 ${\mu}g$/ml의 농도로 처리한 다음, Oil Red O 염색으로 세포 내축적되는 지방구와 지질의 양을 측정한 결과 농도의존적으로 크게 감소됨을 확인하였으며 3T3-L1 지방전구세포의 분화와 관련된 단백질의 발현의 변화를 조사하였다. 지방세포의 특이적 marker인 peroxisome proliferator-activated receptor ${\gamma}$ (PPAR${\gamma}$), cytidine-cytidine-adenosine-adenosine-thymine (CCAAT)/enhancer-binding proteins ${\alpha}$, ${\beta}$ (C/EBP${\alpha}$, C/EBP${\beta}$) 그리고 sterol regulatory element binding protein (SREBP)의 발현이 STME를 처리하였을 때 현저하게 저해됨을 확인하였다. 세포주기의 변화를 분석한 결과 STME는 지방세포 분화초기 단계인 mitotic clonal expansion 단계에서 G1기로 세포주기를 정지시켰다. 더불어 G1 arrest와 관련된 단백질의 변화를 조사 한 결과, 3T3-L1 세포에 STME를 처리하였을 때 p21의 발현량이 확연하게 증가하였으며, Cdk2, E2F-1 그리고 phosphor-Rb의 발현량은 농도의존적으로 감소하였다. 이러한 결과들에 의하여 STME은 메탄올 추출물임에도 불구하고 3T3-L1 지방전구세포가 성숙지방세포로 분화할 때 G1 arrest를 통하여 지방세포 분화를 억제하며 관련 유전자의 발현 억제도 확연하게 확인할 수 있었으며, 이러한 결과는 항 비만 천연물 소재 탐구의 기초자료로 유용하게 쓰일 것으로 사료된다.

• 생명과학회지
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• 제28권7호
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• pp.765-771
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• 2018

$PPAR{\gamma}$는 지방세포의 분화를 조절하는 핵심적인 전사 인자로서 이를 조절하는 후성유전학적 조절 기전이 비만억제 연구에서 중요하게 주목 받고 있다. 선행연구에서 CACUL1이 $PPAR{\gamma}$의 전사 활성 및 지방세포의 분화를 억제하는 corepressor로서 작용함을 밝힌 바 있으며 본 연구에서는 CACUL1의 새로운 결합 단백질로 발굴된 protein arginine methyltransferase 5 (PRMT5)의 $PPAR{\gamma}$ 조절 기능을 분석하였다. PRMT5가 CACUL1과 결합함을 immunoprecipitation assay in vivo와 GST-pull down assay in vitro를 통하여 확인하였다. Luciferase reporter assay 결과로 두 단백질이 상호 협력하여 $PPAR{\gamma}$의 전사 활성을 억제함을 확인하였다. PRMT5가 안정적으로 과발현 또는 knockdown되는 3T3-L1 세포주를 제작하여 지방세포 분화에 미치는 영향을 분석한 결과, PRMT5가 3T3-L1세포의 지방세포 분화를 억제함을 증명하였다. 같은 맥락으로 PRMT5는 $PPAR{\gamma}$의 타겟 유전자인 Lpl과 aP2의 발현을 억제하는 것을 RT-qPCR로 확인하였다. 이상의 연구 결과로 PRMT5이 CACUL1과 결합하여 $PPAR{\gamma}$의 전사 활성을 방해, 나아가 지방세포의 분화를 억제하는 기존에 알려지지 않은 분자적 기전을 처음으로 밝혔다. 따라서, PRMT5 효소 활성의 조절은 비만 억제를 위한 약물 개발에 단서를 제공할 것이다.