• Journal of Life Science
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• v.24 no.5
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• pp.505-514
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• 2014

To examine the anti-obese activity of miscellaneous cereal grains, 80% ethanol extracts from eight selected miscellaneous cereal grains were compared for their cytotoxic effects on 3T3-L1 murine preadipocytes. The ethanol extract of proso millet exhibited the highest cytotoxicity. Further fractionation of the ethanol extract with methylene chloride, ethyl acetate, and n-butanol showed that the cytotoxicity of the ethanol extract was mainly partitioned into the butanol fraction. As compared with differentiated mature adipocytes, 3T3-L1 preadipocytes were more susceptible to the cyctotoxicity of the butanol fraction. When each organic solvent fraction (25 ${\mu}g/ml$) was added during the differentiation period for 6 days, the cell viability was not affected significantly except for the butanol fraction, but the intracellular lipid accumulation declined to a level of 81.5%~50.3% of the control. The Oil Red O staining data also demonstrated that the ethanol extract as well as the butanol fraction could inhibit the differentiation of 3T3-L1 preadipocytes into mature adipocytes. The presence of the butanol extract during the induced adipocytic differentiation also resulted in a significant reduction in the expression levels of critical adipogenesis mediators $(C/EBP{\alpha}$, $PPAR{\gamma}$, aP2, and LPL) to a barely detectable or undetectable level and the cells retained the fibroblast-like morphology of 3T3-L1. In 3T3-L1 cells, the cytotoxicity of the butanol fraction (50-100 ${\mu}g/ml$) was accompanied by mitochondrial membrane potential (${\Delta}{\psi}m$) loss, caspase-3 activation, and PARP degradation. Taken together, these results indicate that proso millet grains possess pro-apoptotic and anti-adipocytic activities toward adipocytes, which can be applicable to prevention of obesity.

• Journal of Applied Biological Chemistry
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• v.60 no.1
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• pp.23-28
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• 2017

Amomum tsao-ko used as a traditional oriental herbal medicine, is indigenous to several Asia countries. In this study, we investigated anti-obesity activity of the ethanol extract of Amomum Taso-ko (A. tsao-ko). The ethanol extract of A. tsao-ko inhibited adipocyte differentiation using Oil Red O assay in 3T3-L1 cells. Inhibitory effect of the ethanol extract of A. tsao-ko on adipogenesis was modulated by down-regulation adipogenic transcriptional factor such as peroxisome proliferator-activated receptor ${\gamma}$ ($PPAR{\gamma}$), CCAAT-enhancer-binding protein ${\alpha}$ ($C/EBP{\alpha}$) and suppressed expression of fatty acid synthase, aP2, and resistin. We demonstrated that A. tsao-ko significantly inhibited adipogenesis and reduced $PPAR{\gamma}$ and $C/EBP{\alpha}$ expression in a dose-dependent manner. These results suggest that A. tsao-ko has an anti-obesity effect by inhibition of adipogenic transcription factor and adipocyte-specific genes in 3T3-L1 cells.

• Journal of Life Science
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• v.31 no.1
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• pp.1-9
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• 2021

Brown seaweeds have been shown to decrease blood glucose levels and improve insulin sensitivity previously. In this study, we investigated the effect of fucoidan, a complex polysaccharide derived from brown seaweeds, on glucose uptake to improve insulin resistance, and examined its mechanism of action in 3T3-L1 adipocytes. We observed that fucoidan significantly increased glucose uptake and it was related to an increased expression of plasma membrane-glucose transporter 4 (PM-GLUT4) in 3T3-L1 adipocytes. Fucoidan treatment increased the activation of phosphatidylinositol-3-kinase (PI3K) and the phosphorylation of insulin receptor substrate 1 (IRS1tyr) compared with that of the control cells. Fucoidan also promoted the phosphorylation of Akt and protein kinase C (PKC)-λ/ζ compared to that of the control cells. Moreover, fucoidan significantly upregulated acetyl-CoA-carboxylase (ACC) and adenosine monophosphate - activated protein kinase (AMPK) phosphorylation. As a result, translocation of GLUT4 was significantly enhanced in 3T3-L1 adipocytes, which significantly promoted glucose uptake via the PI3K/AMPK pathways. The elevation of glucose uptake by fucoidan was blocked by inhibitor of PI3K and inhibitor of AMPK in 3T3-L1 adipocytes. These findings indicate that fucoidan might ameliorate glucose uptake through GLUT4 translocation to the plasma membrane by activating the PI3K/Akt and AMPK pathways in 3T3-L1 adipocytes. Fucoidan is thought to be of high material value to diabetes treatments and functional foods.

• Journal of Life Science
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• v.21 no.9
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• pp.1346-1353
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• 2011

Sophora tonkinensis Gapnep has been used as a traditional herbal medicine in oriental regions since ancient times. In this study, the effect and mechanism of the MeOH extract of Sophora tonkinensis Gapnep (STME) on adipocite differentiation and adipogenesis in 3T3-L1 preadipocites were investigated. Treatment with STME in the concentration range of 0-200 ${\mu}g$/ml significantly inhibited the differentiation of 3T3-L1 preadipocites in a dose-dependent manner, as determined by a decrease in intracellular lipid droplets and lipid contents measured by Oil Red O staining. In association with the inhibitory effect of lipid accumulation, the expressions of the proteins concerned with adipogenesis in 3T3-L1 preadipocites were also investigated. Treatment with STME reduced the expressions of peroxisome proliferator-activated receptor ${\gamma}$ (PPAR${\gamma}$), cytidine-cytidine-adenosine-adenosine-thymine (CCAAT)/enhancer-binding proteins ${\alpha}$ and ${\beta}$ (C/EBP${\alpha}$ and C/EBP${\beta}$) and sterol regulatory element binding protein (SREBP), which are adipocyte specific markers. In flow cytometry analysis, the inhibitory effect of differentiation was caused by G1 arrest and following mitotic clonal expansion cease. Therefore, we also investigated the alteration of G1 phase arrest-related proteins. As a result, the expression of p21 protein was significantly increased, while the expressions of Cdk2, E2F-1 and phospho-Rb were reduced in a dose-dependent manner in STME treated 3T3-L1 cells. According to these results, STME might inhibit differentiation through G1 arrest in 3T3-L1 preadipocytes adipogenesis, and further studies, which are in progress, have to be completed to identify the active compounds.

• Korean Journal of Clinical Laboratory Science
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• v.51 no.4
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• pp.514-520
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• 2019

This study investigated the improved lipid metabolism effect of 3T3-L1 cells induced by adipocytes using the dichloromethane (DCM) fraction in the organic solvent extract of Wassong (Orostachys japonicus). To confirm the cell cytotoxicity, each of 6 fractions of organic solvent extracts (EtOH, Hexane, DCM, EtOAc, BuOH, and H₂O) was examined using MTS assay. As a result, it was confirmed that the DCM extract was stable over the whole range of concentrations, and a DCM fraction was used to confirm the improved lipid metabolism effect. Lipid excretion was measured to confirm the change of lipid metabolism. 3T3-L1 cells induced by adipocytes were treated with DCM extract and stained with oil-red O to evaluate lipid accumulation. As a result, it was confirmed that the lipid efflux was significantly improved. In order to confirm the mechanism of lipid efflux, the mRNA expressions of ABCA1 and ABCG1, which are lipid transport proteins, were confirmed by real-time PCR. Therefore, the present study confirmed that the DCM extract from Orostachys japonicus has the effect of improving the lipid metabolism on 3T3-L1 adipocytes. In addition, the results of this study will be used as the basis for the development of functional foods using Orostachys japonicus and also for conducting research on the detailed mechanisms.

• Journal of Life Science
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• v.22 no.8
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• pp.1052-1056
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• 2012

Undaria pinnatifada has been used as a natural diet food with few calories and as a source of iodine. Even though U. pinnatifida has been regarded as a diet food, the mechanisms of its inhibitory effects on adipocyte differentiation and the accumulation of fat in adipocytes are poorly understood. In this study, the effect and mechanism of U. pinnatifida ethanol extract on 3T3-L1 differentiation into adipocytes were investigated. The effects of U. pinnatifida ethanol extract on cell viability and the anti-adipogenic effect were investigated via MTT assay, Oil red O staining, RT-PCR, and western blot. The U. pinnatifida ethanol extract did not show toxicity up to a concentration of 50 ${\mu}g/ml$. The addition of U. pinnatifida ethanol extract decreased triglyceride contents by 40% when 50 ${\mu}g/ml$ of U. pinnatifida ethanol extract was added during 3T3-L1 differentiation and adipocyte triglyceride formation. The transcription and expression of peroxisome proliferator-activated receptor ${\gamma}$ ($PPAR{\gamma}$), leptin, and hormone-sensitive lipase (HSL) as adipocyte-specific proteins were determined by RT-PCR and western blot. The overexpression of $PPAR{\gamma}$ could accelerate adipocyte differentiation. Also, leptin was secreted for triglyceride accumulation in the adipocytes and the increase of adipocyte cell size. Thus, $PPAR{\gamma}$ and leptin were used as indicators of obesity. $PPAR{\gamma}$ and leptin were repressed by the increased addition of U. pinnatifida ethanol extract. This indicates that U. pinnatifida was effective as an anti-obesity agent by repressing the differentiation of 3T3-L1 into adipocytes and inhibiting triglyceride formation in adipocytes.

• Journal of Life Science
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• v.32 no.7
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• pp.523-531
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• 2022

The purpose of this study was to evaluate the effect of antioxidant and anti-adipogenic activities in ethanol extracts from herb mixture (Ephedra sinica, Atractylodes lance, Gypsum fibrosum, and Theobroma cacao). DPPH, ABTS⁺ radical and xanthine oxidase scavenging activities were measured for antioxidant activity. Extracts of the herb mixture had 75.0, 100.8, and 79.5% scavenging activities at 1,000 ㎍/ml concentration, respectively. We investigated the inhibition of adipogenesis and adipocyte differentiation with an extract of an herb mixture in 3T3-L1 preadipocytes. An extract from the herb mixture at concentrations between 0 and 50 ㎍/ml did not affect 3T3-L1 cell viability. Treatment with herb mixture extracts of 25, 50, and 75 ㎍/ml in 3T3-L1 preadipocytes inhibited lipid accumulation in a dose-dependent manner. As a result of a Western blot experiment, it was shown that the herb mixture inhibited the differentiation transcription factors, PPARγ and C/EBPa, by 44.2 and 77.6%, respectively, at a concentration of 75 ㎍/ml in MDI-induced differentiated 3T3-L1 cells. As a result of RT-PCR, the gene expression of C/EBPa, SREBP-1c, and PPARγ was significantly inhibited by 43.4%, 59.6%, and 55.3%, respectively, at the concentration of 75 ㎍/ml of the herb mixture compared with the MDI-treated group. In addition, the expression of fatty acid synthase (FAS), a fatty acid synthesis regulator, was suppressed. These results can be applied to develop a functional food for anti-obesity with a herb mixture.

• Journal of Life Science
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• v.14 no.6 s.67
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• pp.967-974
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• 2004

The osteoblastic cell activity is important for born formation, thus, this study was performed to investigation of that the effect of edible sources, Rubus coreanus Miquel (RCM), on the proliferation and differentiation of MC3T3-E1 osteoblastic like cell. The effects of RCM extract on cell proliferation were measured by MIT assay. At 1, $10\;{\mu}g/mL$ of RCM extract treated, that were elevated of cell proliferation to 103 and $142\%$ via control, respectively. And the cell differentiation were measured as alkaline phosphatase (ALP) activity at 3, 9, 18, and 27 days. As the results, the $10\;{\mu}g/mL$ was increased ALP activity more than 2.6 times compared with control, 1.4 times via positive control at 27th day (p<0.05). The optical concentration of RC extract was rechecked by ALP staining and Alizarin Red staining for investigation of the induction of ALP activity, nodule formation by mineralization. mRNA expression analysis showed that the RCM $(10\;{\mu}g/mL)$ increased in SOX9 as well as ALP in MC3T3-E1 cells. These results suggest that RC extract was stimulates the MC3T3-E1 cell proliferation and differentiation.

• Journal of Applied Biological Chemistry
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• v.62 no.4
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• pp.417-424
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• 2019

The aim of this study is to investigate the effect of Lespedeza maximowiczii var. tricolor Nakai (LMTN) on glucose metabolism. LMTN extract significantly enhanced the glucose uptake and lipid accumulation in 3T3-L1 adipocytes compared with control. Also, LMTN extract in 3T3-L1 adipocytes significantly increased the protein expression of peroxisome proliferator-activated receptor (PPAR)γ, insulin receptor substrate-1, and glucose transporter (GLUT)4. The regulatory effect on glucose uptake or insulin signal transduction of LMTM extract was lower than troglitazone or pinitol such as the positive control, but increased PPARγ activation. Additionally, LMTM extract has an insulin-mimetic effect. In db/db mice, LMTN extract (250 mg/kg BW) significantly reduced water and food intake, blood glucose, and level of plasma triglyceride and total cholesterol. Furthermore, the expression of PPARã and GLUT4 mRNA in adipose or muscle tissue effectively was increased by oral treatment of LMTN extract. Thus, our results suggest that LMTN extract improves the glucose metabolism through PPARγ and insulin-mimetic effect in 3T3-L1 adipocytes and db/db mice.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.6
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• pp.837-843
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• 2013

Sasa borealis is a major source of bamboo leaves used for traditional medicine in Korea. Obesity is a serious health problem in industrialized countries that has been implicated in various diseases, including type 2 diabetes, hypertension, cancer, and coronary heart disease. Recent reports have proposed mechanisms to reduce obesity by decreasing preadipocyte differentiation, and proliferation in 3T3-L1 preadipocyte. The preadipocytes play a key role by differentiation into mature adipocytes and increasing fat mass. In this study, we investigated whether ethanol-soluble extracts and ethyl acetate-soluble fractions from Sasa borealis inhibits intracellular accumulation of lipid droplets in a dose-dependent manner in 3T3-L1 cells (an important model system for studying adipogenesis). The down-regulation of PPAR${\gamma}$ and C/EBP${\alpha}$ (key adipogenic transcription factors) were confirmed by the reverse transcription polymerase chain reaction (RT-PCR). Ethyl acetate-soluble fractions from Sasa borealis attenuated the expression of PPAR${\gamma}$ and C/EBP${\alpha}$. These results suggest that Sasa borealis inhibits adipogenic differentiation by regulating adipogenic transcription factors in 3T3-L1 cells. Therefore, Sasa borealis extracts may be a good candidate for the management of obesity.