• Molecules and Cells
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• 제45권12호
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• pp.963-975
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• 2022

Exogenous polyamines are able to induce life span and improve glucose homeostasis and insulin sensitivity. However, the effects of exogenous polyamines on adipocyte differentiation and which polyamine transporters mediate them have not been elucidated yet. Here, we identified for the first time that exogenous polyamines can clearly stimulate adipocyte differentiation through polyamine transporters, solute carrier family 3 member A2 (SLC3A2) and SLC7A1. Exogenous polyamines markedly promote 3T3-L1 adipocyte differentiation by increasing the intracellular lipid accumulation and the expression of both adipogenic and lipogenic genes in a concentration-dependent manner. In particular, exogenous putrescine mainly regulates adipocyte differentiation in the early and intermediate stages. Moreover, we have assessed the expression of polyamine transporter genes in 3T3-L1 preadipocytes and adipocytes. Interestingly, the putrescine-induced adipocyte differentiation was found to be significantly suppressed in response to a treatment with a polyamine transporter inhibitor (AMXT-1501). Furthermore, knockdown experiments using siRNA that specifically targeted SLC3A2 or SLC7A2, revealed that both SLC3A2 and SLC7A2 act as important transporters in the cellular importing of exogenous putrescine. Thus, the exogenous putrescine entering the adipocytes via cellular transporters is involved in adipogenesis through a modulation of both the mitotic clonal expansion and the expression of master transcription factors. Taken together, these results suggest that exogenous polyamines (such as putrescine) entering the adipocytes through polyamine transporters, can stimulate adipogenesis.

• 생명과학회지
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• 제22권8호
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• pp.1052-1056
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• 2012

미역(Undaria pinnatifada)은 낮은 칼로리 및 요오드의 원료로써 천연체중조절식품으로 알려져 있다. 미역이 체중조절식품으로 알려져 있음에도 불구하고, 지방세포 분화 및 지방축적에 관한 저해 기작은 연구가 미비하다. 본 연구에서는 3T3-L1에서 지방세포로 분화가 일어나는 단계에서 미역에탄올추출물의 효과 및 기작을 확인하였다. 미역에탄올추출물의 독성과 지방축적저해효과는 MTT assay, Oil red O staining, RT-PCR과 western blot으로 분석하였다. 미역에탄올추출물은 50 ${\mu}g/ml$의 농도에서 독성을 띄지 않았다. 3T3-L1의 분화 및 지방세포에서 triglyceride축적과정동안 50 ${\mu}g/ml$의 미역에탄올추출물을 처리하였으며, 미역에탄올추출물은 지방세포에서 triglyceride의 축적을 40% 감소시켰다. 지방세포 특이적 단백질인 Peroxisome proliferator activated receptor ${\gamma}$ ($PPAR{\gamma}$), leptin과 Hormone sensitive lipase (HSL)의 발현은 RT-PCR과 western blot으로 확인하였다. $PPAR{\gamma}$의 과발현은 지방세포의 분화를 촉진시킨다. 또한 지방세포 크기의 증가와 세포 내 triglyceride의 함량에 따라 leptin은 세포 외로 분비된다. 그러므로 $PPAR{\gamma}$와 leptin은 비만의 지표로 사용된다. 첨가한 미역에탄올추출물의 농도가 높아질수록 $PPAR{\gamma}$와 leptin의 발현이 억제되었다. 이상의 결과를 통하여, 미역의 에탄올 추출물은 지방전구세포의 분화를 억제시키며, 지방세포 내 triglyceride의 축적을 저해하는 것으로 판단된다.

• Food Science and Biotechnology
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• 제14권4호
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• pp.514-517
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• 2005

Abilities of Korean black raspberry (KBR) wine to induce differentiation of 3T3 L1 adipocytes and express HepG2 cardiovascular disease-related genes were determined and compared with those of red wine. Red wine attenuated the differentiation of adipocytes faster than KBR wine and control. KBR wine decreased cholesterol concentration in HepG2 cells at a similar level to that of red wine, resulting in similar degrees of suppression in apolipoprotein B100, and enhancement in LDL receptor and 3-hydroxy-3-methylglutaryl coenzyme A reductase mRNA levels.

• 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.

• Nutrition Research and Practice
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• 제5권3호
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• pp.192-197
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• 2011

The dietary intake of whole grains is known to reduce the incidence of chronic diseases such as obesity, diabetes, cardiovascular disease, and cancer. To investigate whether there are anti-adipogenic activities in various Korean cereals, we assessed water extracts of nine cereals. The results showed that treatment of 3T3-L1 adipocytes with Sorghum bicolor L. Moench, Setaria italica Beauvois, or Panicum miliaceum L. extract significantly inhibited adipocyte differentiation, as determined by measuring oil red-O staining, triglyceride accumulation, and glycerol 3-phosphate dehydrogenase activity. Among the nine cereals, P. miliaceum L. showed the highest anti-adipogenic activity. The effects of P. miliaceum L. on mRNA expression of peroxisome proliferator-activated receptor-${\gamma}$, sterol regulatory element-binding protein 1, and the CCAAT/enhancer binding protein-${\alpha}$ were evaluated revealing that the extract significantly decreased the expression of these genes in a dose-dependent manner. Moreover, P. miliaceum L. extract changed the ratio of monounsaturated fatty acids to saturated fatty acids in adipocytes, which is related to biological activity and cell characteristics. These results suggest that some cereals efficiently suppress adipogenesis in 3T3-L1 adipocytes. In particular, the effect of P. miliaceum L. on adipocyte differentiation is associated with the downregulation of adipogenic genes and fatty acid accumulation in adipocytes.

• 대한한의학회지
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• 제23권1호
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• pp.83-91
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• 2002

Objectives : In this study, water extracts from Coix lachryma-jobi Linne var. mayuen Stapf. were investigated for their effects on insulin-like action and glucose uptake in 3T3-Ll cells. Methods : We examined the effects of insulin-like action on the differentiation of 3T3-Ll fibroblasts. The Coicis Semen was treated with hot water and the extract was freeze-dried. The hot water extract was chromatographed on nonionic polymer resin (Amberlite XAD-4, Sigma) with distilled water (Fr. 1), 20% (Fr. 2), 40% (Fr. 3), 60% (Fr. 4), 80% (Fr. 5), and 100% EtOH (Fr. 6), successively. Results : Total extract of Coicis Semen was fractionated into 0 to 100% MeOH with Amberlite XDA-4 column. Treatment of cells with $10{\;}\mu\textrm{g}/ml$ of total extracts of Coicis Semen significantly increased the differentiation (p<0.05). At $1{\;}\mu\textrm{g}/ml$ level of insulin, the differentiation was accelerated (p<0.01). The effect of extracts plus insulin on the differentiation was greater than that of insulin alone. In 3T3-Ll adipocytes, glucose uptake was higher by 2.7 times with 5 uM of total extract in low dosage of insulin (3 ng/ml) than without total extract. 5 and 50 uM of water and 40% MeOH fractions increased glucose uptake by 3.5 times in 3T3-Ll adipocytes (p<0.00l). Conclusions : Coicis Semen contains compounds which playa role of insulin-like action and insulin sensitizer.

• Animal Bioscience
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• 제37권5호
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• pp.929-943
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• 2024

Objective: The present study was executed to explore the molecular mechanism of fibroblast growth factor 10 (FGF10) gene in bovine adipogenesis. Methods: The bovine FGF10 gene was overexpressed through Ad-FGF10 or inhibited through siFGF10 and their negative control (NC) in bovine adipocytes, and the multiplicity of infection, transfection efficiency, interference efficiency were evaluated through quantitative real-time polymerase chain reaction, western blotting and fluorescence microscopy. The lipid droplets, triglycerides (TG) content and the expression levels of adipogenic marker genes were measured during preadipocytes differentiation. The differentially expressed genes were explored through deep RNA sequencing. Results: The highest mRNA level was found in omasum, subcutaneous fat, and intramuscular fat. Moreover, the highest mRNA level was found in adipocytes at day 4 of differentiation. The results of red-oil o staining showed that overexpression (Ad-FGF10) of the FGF10 gene significantly (p<0.05) reduced the lipid droplets and TG content, and their down-regulation (siFGF10) increased the measurement of lipid droplets and TG in differentiated bovine adipocytes. Furthermore, the overexpression of the FGF10 gene down regulated the mRNA levels of adipogenic marker genes such as CCAAT enhancer binding protein alpha (C/EBPα), fatty acid binding protein (FABP4), peroxisome proliferator-activated receptor-γ (PPARγ), lipoprotein lipase (LPL), and Fas cell surface death receptor (FAS), similarly, down-regulation of the FGF10 gene enriched the mRNA levels of C/EBPα, PPARγ, FABP4, and LPL genes (p<0.01). Additionally, the protein levels of PPARγ and FABP4 were reduced (p<0.05) in adipocytes infected with Ad-FGF10 gene and enriched in adipocytes transfected with siFGF10. Moreover, a total of 1,774 differentially expressed genes (DEGs) including 157 up regulated and 1,617 down regulated genes were explored in adipocytes infected with Ad-FGF10 or Ad-NC through deep RNA-sequencing. The top Kyoto encyclopedia of genes and genomes pathways regulated through DEGs were the PPAR signaling pathway, cell cycle, base excision repair, DNA replication, apoptosis, and regulation of lipolysis in adipocytes. Conclusion: Therefore, we can conclude that the FGF10 gene is a negative regulator of bovine adipogenesis and could be used as a candidate gene in marker-assisted selection.

• 약학회지
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• 제55권4호
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• pp.309-313
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• 2011

Abnormal growth of adipocyte characterized by increased cell numbers and differentiation is considered as an major pathological characteristic feature in obesity. Thus, inhibition of mitogenesis of preadipocytes and their differentiation to adipocytes would be beneficial for the prevention and inhibition of obesity. In the present study, we attempted to evaluate anti-adipogenic activity of eggplants (the fruits of Solanum melongena L.) employing preadipocytes cell line, 3T3-L1 as an in vitro assay system. Water extract of eggplants significantly inhibited adipocyte differentiation when treated during adipocyte differentiation process, as assessed by measuring fat accumulation using Oil Red O staining. Eggplant extract, however, showed little effects on fully differentiated adipocytes. Eggplant didn't show significant toxicity up to 500 ${\mu}g$/ml to the 3T3-L1 cells. Further studies with interval treatment demonstrated that eggplant exerted inhibitory activity on adipocyte differentiation via acting on early stage of adipogenesis. Conclusively, eggplants are suggested to be beneficial for the prevention of obesity.

• 대한의생명과학회지
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• 제18권3호
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• pp.291-298
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• 2012

The antiobesity effect of the mixture of black garlic and Garsinia cambogia extracts (BGG) was investigated by measuring the Oil red O staining and the expressions of adipogenic genes during preadipocyte differentiation by real-time PCR in the 3T3-L1 adipocytes. BGG reduced contents of Oil red O dye in the 3T3-L1 adipocytes. mRNA expression levels of SREBP1c, C/EBPa, aP2/FABP4, and $PPAR{\gamma}$ which are adipogenic transcription factor, in cells treated with BGG were also significantly down regulated. Also, the phosphorylation of AMP-activated protein kinase (AMPK) in L6 cells was more increased by BGG. These results indicate that BGG seems to be more attractive compound for application of industry than individual extracts such as black garlic and Garsinia cambogia, considering it has two effects not only inhibit the preadipocyte differentiation but also activate the phosphorylation of AMPK unlike other two compound.

• Natural Product Sciences
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• 제16권4호
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• pp.291-294
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• 2010

In obesity, adipocytes undergo abnormal growth characterized by increased cell numbers and differentiation. Thus, inhibition of mitogenesis of preadipocytes and their differentiation to adipocytes would be beneficial for the prevention and progression of obesity. In the present study, we attempted to evaluate antiadipogenic activity of adlay (Coix seed, the seed of Coix lacryma-jobi L. var. ma-yuen Stapf) employing preadipocytes cell line, 3T3-L1 as an in vitro assay system. Because several varieties of adlay are in use in Korea, anti-adipogenic activity of three varieties of adlay such as Sang-Gang, Jo-Hyun and Yulmu-Ilho was evaluated. All the three varieties of adlay showed significant inhibitory activity on adipocyte differentiation as assessed by measuring fat accumulation using Oil Red O staining. Adlay, however, showed little effects on adipocyte proliferation. Further studies with interval treatment demonstrated that adlay exerted inhibitory activity on adipocyte differentiation via acting on early stage of adipogenesis. Taken together, adlay might be useful in the prevention of obesity.