• Journal of Animal Science and Technology
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• v.66 no.1
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• pp.204-218
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• 2024

Elsholtzia fruticosa (EF) is present in tropical regions throughout South Asian countries as well as the Himalayas. Although it has been used as a traditional medicine to treat digestive, respiratory, and inflammatory issues, its effect on preadipocyte differentiation is unknown. In this study, we examined the effects of a methanol extract prepared from EF on the differentiation of 3T3-L1 preadipocytes. Cell differentiation was assessed by microscopic observation and oil-red O staining. The expression of adipogenic and lipogenic genes, including PPARγ and C/EBPα, was measured by western blot analysis and quantitative real-time polymerase chain reaction (qRT-PCR), to provide insight into adipogenesis and lipogenesis mechanisms. The results indicated that EF promotes the differentiation of 3T3-L1 preadipocytes, with elevated lipid accumulation occurring in a concentration-dependent manner without apparent cytotoxicity. EF enhances the expression of adipogenic and lipogenic genes, including PPARγ, FABP4, adiponectin, and FAS, at the mRNA and protein levels. The effect of EF was more pronounced during the early and middle stages of 3T3-L1 cell differentiation. Treatment with EF decreased C/EBP homologous protein (CHOP) mRNA and protein levels, while increasing C/EBPα and PPARγ expression. Treatment with EF resulted in the upregulation of cyclin E and CDK2 gene expression within 24 h, followed by a decrease at 48 h, demonstrating the early-stage impact of EF. A concomitant increase in cyclin-D1 levels was observed compared with untreated cells, indicating that EF modulates lipogenic and adipogenic genes through intricate mechanisms involving CHOP and cell cycle pathways. In summary, EF induces the differentiation of 3T3-L1 preadipocytes by increasing the expression of adipogenic and lipogenic genes, possibly through CHOP and cell cycle-dependent mechanisms.

• The Korea Journal of Herbology
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• v.39 no.2
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• pp.27-33
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• 2024

Objective : Obesity is a disease caused by energy imbalance and increases the risk of complications such as cardiovascular disease and cancer. Additionally, a recent study reported that excessive production of ROS stimulates the expression of transcription factors related to adipogenesis, such as PPAR-𝛾, C/EBP-𝛼 and 𝛽, in 3T3-L1 cells. In this study, Paeonia anomala L. (PAL) was selected as a candidate for improving and preventing obesity from Mongolian medical literature, and its effect on fat metabolism was observed in vitro. Methods : The activity of PAL extract against lipid accumulation was measured in 3T3-L1 cells through Oil red O staining. Additionally, the expression of lipid metabolism-related proteins was measured by western blot. Results : Intracellular lipid accumulation was significantly lower in the treatment group than in the control group. In addition, pAMPK/AMPK protein expression in the treatment group tended to increase in a concentration-dependent manner, and PPAR-𝛾 protein expression in the treatment group showed a significant decrease in a concentration-dependent manner compared to the control group. Conclusion : These results suggest that PAL extract may have an anti-obesity effect, and it is believed that it can be used to develop functional materials for the treatment of obesity.

• IMMUNE NETWORK
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• v.20 no.5
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• pp.42.1-42.10
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• 2020

Long-lasting post-switched plasma cells (PCs) arise mainly from germinal center (GC) reactions, but little is known about the mechanism by which GC B cells differentiate into PCs. Based on our observation that the expression of the transcription factor CCAAT/enhancer binding protein β (C/EPBβ) is associated with the emergence of post-switched PCs, we enquired whether a cell-autonomous function of C/EPBβ is involved in the program for PC development. To address this, we generated C/EPBβ-deficient mice in which the Cebpb locus was specifically deleted in B cells after transcription of the Ig γ1 constant gene segment (Cγ1). In response to in vitro stimulation, B cells from these Cebpb^fl/flCγ1^Cre/+ mice had defects in the induction of B lymphocyte-induced maturation protein 1 (Blimp1) and the formation of IgG1⁺ PCs, but not in proliferation and survival. At steady state, the Cebpb^fl/flCγ1^Cre/+ mice had reduced serum IgG1 titers but normal IgG2c and IgM titers. Moreover, upon immunization with T-dependent Ag, the mice produced reduced levels of Ag-specific IgG1 Ab, and were defective in the production of Ag-specific IgG1 Ab-secreting cells. These results suggest that a cell-autonomous function of C/EPBβ is crucial for differentiation of post-switched GC B cells into PCs through a Blimp1-dependent pathway.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.3
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• pp.279-288
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• 2017

This study investigated the manufacturing of fermented soybean oil using a fermenting strain commonly processed for soybeans [Bacillus amyloliquefaciens (BA), Bacillus subtilis (BS), Lactobacillus acidophilus (LBA), and B. subtilis+L. acidophilus (BLO)] and evaluated its anti-obesity activities. Cytotoxicity of four kinds of fermented soybean oils was not observed in 3T3-L1 preadipocytes at 10 and $50{\mu}g/mL$. Triglyceride content was reduced by 20.6% in the BLO group at a treatment concentration of $50{\mu}g/mL$. The simultaneous treatment of fermented soybean oil and differentiation induction medium decreased $PPAR{\gamma}$ and $C/EBP{\alpha}$ gene expression at a concentration of $50{\mu}g/mL$ and blocked adipocyte differentiation by increasing adiponectin gene expression. The inhibitory effect of adipocyte differentiation was greatest in the BLO group. C57BL/6J mice were examined for 4 weeks after being separated into seven groups [normal diet group (N), high fat diet group (C), group fed high fat diet combined with regular soybean oil (SO), group fed non-fermented soybean oil (NF), and groups fed high fat diet combined with 5% fermented soybean oil (BA, BS, LBA, and BLO)] to identify the effects of soybean oil on body weight, serum lipid, adiponectin, insulin, and leptin levels in mice with high fat diet-induced obesity. The body weight and serum lipid level of the C group increased drastically compared to those of the N group. In contrast, the group fed a diet combined with fermented soybean oil showed decreases in weight, serum total cholesterol, LDL-cholesterol, and triglyceride levels compared to those of the C group. Moreover, soybean oil was found to be effective in the BLO group. In conclusion, fermented soybean oil has positive effects in prohibiting adipocyte differentiation increased by high fat diet and improving serum lipid composition. Therefore, fermented soybean oil can be used as a functional food material with anti-obesity activity.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.12
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• pp.1701-1707
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• 2016

The present study investigated the anti-obesity effect of pine cone (PC, Pinus koraiensis) supercritical extract in high-fat diet (HFD)-induced obese mice. Male C57BL/6J mice were treated with HFD, HFD+catechin, and HFD+PC [two different doses, 20 mg/kg body weight (b.w.) and 100 mg/kg b.w.] in each AIN93G supplement for 8 weeks. Treatment of HFD mice with both low and high doses of PC significantly reduced body weight gain compared to HFD mice. Liver weight of mice was reduced in both the low and high dose PC-supplemented groups (24.19% and 19.83%, respectively). Total adipose tissue weight of mice was reduced in both the low and high dose PC-supplemented groups (45.54% and 62.66%, respectively). Serum total cholesterol, triglyceride, LDL cholesterol, and HDL cholesterol were reduced in the low and high dose PC-supplemented groups, and ratios of HDL cholesterol to LDL cholesterol increased by 94.55% in the high dose PC-supplemented group. Serum leptin was significantly reduced in the low and high dose PC-supplemented groups (28.14% and 62.72%, respectively). These results were supported by genetic expression of protein and enzymes related to lipid metabolism assessed by real-time PCR. There was significant reduction of lipid regulatory transcription factors such as $PPAR-{\gamma}$, C/EBP, and SREBP and lipid enzymes such as fatty acid synthesis and lipoprotein lipase in the low and high dose PC-supplemented groups. However, there was no statistical difference between low and high dose PC treatments. These results suggest that pine cone supercritical extract supplementation is able to regulate serum lipid profiles by reducing total cholesterol, triglyceride, and LDL cholesterol levels, followed by regulation of expression of lipid metabolic factors, resulting in reduction of weight gain in HFD-induced obese mice.

• 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 the Korean Society of Food Science and Nutrition
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• v.43 no.10
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• pp.1477-1483
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• 2014

This study investigated the anti-obesity effects of African mango (Irvingia gabonesis, IGOB $131^{TM}$) extract in leptin-deficient obese mice. Experimental groups were treated with two different doses of IGOB $131^{TM}$ (1% and 2% in each AIN93G supplement) for 8 weeks. Treatment of obese mice with both low and high dose of IGOB $131^{TM}$ significantly reduced body weight gain by 10.9% and 13.3%, respectively, compared to control obese mice. Subcutaneous adipose tissue weight of mice was significantly reduced by 18% by low-dose and 23% by high-dose supplementation. This result was supported by micro-CT analysis around the abdominal regions of mice, indicating that the adipose tissue area and volume were significantly reduced by treatment with IGOB $131^{TM}$. Serum levels of triglycerides in the low- and high-dose groups were reduced by 36.5% and 43.8%, respectively, upon treatment with IGOB $131^{TM}$, whereas total cholesterol levels were reduced by 31.8% and 35.4%. Interestingly, the serum LDL level decreased upon treatment with IGOB $131^{TM}$ while the serum level of HDL dramatically increased upon high-dose treatment with IGOB $131^{TM}$, resulting in a significant reduction in the LDL to HDL ratio of 59.2%. These results were supported by the expression levels of enzymes and proteins related to lipid metabolism assessed by real-time PCR. There was a significant increase of in adiponectin expression as well as significant decreases in the expression of FAS, LPL, and lipid regulatory transcription factors such as PPAR-${\gamma}$, C/EBP, and SREBP upon both low- and high-dose IGOB $131^{TM}$ treatment. However, there was no statistical difference between low- and high-dose treatments. These results suggest that IGOB $131^{TM}$ is able to regulate the serum lipid profiles by reducing triglyceride and increasing HDL levels as well as regulate expression of lipid metabolic factors, resulting in reduction of a weight gain in leptin-deficient obese mice.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.8
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• pp.1079-1085
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• 2012

Obesity is an important issue worldwide as it may associated with increased prevalence of metabolic diseases. Mung bean is known as a functional food for decreasing the glycemic index and lipid profile of plasma. The purpose of this study was to investigate the anti-obesity effects of vitexin from mung bean on the regulation of adipocyte differentiation and adipocytokine secretion. When 3T3-L1 adipocytes were treated with vitexin from days 0 to 14 at various levels of 25, 50, 100, and $200{\mu}M$, there was no change in cell viability. Vitexin treatment at 50, 100, and $200{\mu}M$ decreased triacylglycerol levels in cells, but only $100{\mu}M$ vitexin induced lipolysis. At $200{\mu}M$ of vitexin, phosphorylation of p38 and ERK, which causes secretion of inflammatory adipocytokines, was depressed, whereas there was an increase in expression of $PPAR{\gamma}$, the key regulator of adipocyte differentiation. Phosphorylation of AMPK increased at $100{\mu}M$ vitexin. TNF-${\alpha}$ and aP2 mRNA expression increased at $25{\mu}M$ vitexin, whereas only TNF-${\alpha}$ mRNA expression increased at $200{\mu}M$ vitexin. Further, the mRNA levels of TNF-${\alpha}$ and aP2 decreased at other concentrations in a dose-dependent manner. Since we observed that mRNA expression of C/EBP, SREBP1, and $PPAR{\gamma}$ did not change upon vitexin treatment, our future studies will investigate other genes such as mTOR, which is related with apoptosis signaling, or SIRT1, which is associated with inhibition of adipogenesis. Our results indicate that vitexin at concentrations between 100 and $200{\mu}M$ is suitable in vivo for the development of mung bean as an anti-obesity therapy or functional food.

• Food Science and Preservation
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• v.21 no.1
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• pp.97-106
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• 2014

This study was performed in order to investigate the anti-obesity effect of Polygala tenuifolia on lipid mechanism in 3T3-L1 adipocytes. The chemical composition of the P. tenuifolia was analyzed in order to assess its nutritional value. Total dietary fiber was the highest among the proximate component of the P. tenuifolia. These results showed that the P. tenuifolia may be used as a potential functional ingredient for anti-obesity effect. Intracellular lipid droplets in the adipocyte were stained with oil-red O dye and quantified. In comparison to the control, lipid accumulation was significantly decreased by 40.1% and 22.4% when treated with the water extract and 70% EtOH extract of the P. tenuifolia at the concentration of $10{\mu}g/mL$, respectively. The anti-adipogenic effect of the water extract was stronger than that of the 70% EtOH extract. The gene expression levels were measured via Western blot and real-time PCR. As a result, the water extract was found to have decrease the gene expression of SREBP-1c, PPAR, $C/EBP{\alpha}$, FAS, ACC in a dose-dependent manner. These indicate that the water extract inhibits pre-adipocyte differentiation and adipogenesis by blocking the SREBP-1c gene expression in 3T3-L1 cells. Therefore, P. tenuifolia can be used as an effective anti-obesity agent.

• Journal of Ginseng Research
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• v.41 no.3
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• pp.257-267
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• 2017

Background: Heat-processed ginseng, sun ginseng (SG), has been reported to have improved therapeutic properties compared with raw forms, such as increased antidiabetic, anti-inflammatory, and antihyperglycemic effects. The aim of this study was to investigate the antiobesity effects of SG through the suppression of cell differentiation and proliferation of mouse 3T3-L1 preadipocyte cells and the lipid accumulation in Caenorhabditis elegans. Methods: To investigate the effect of SG on adipocyte differentiation, levels of stained intracellular lipid droplets were quantified by measuring the oil red O signal in the lipid extracts of cells on differentiation Day 7. To study the effect of SG on fat accumulation in C. elegans, L4 stage worms were cultured on an Escherichia coli OP50 diet supplemented with $10{\mu}g/mL$ of SG, followed by Nile red staining. To determine the effect of SG on gene expression of lipid and glucose metabolism-regulation molecules, messenger RNA (mRNA) levels of genes were analyzed by real-time reverse transcription-polymerase chain reaction analysis. In addition, the phosphorylation of Akt was examined by Western blotting. Results: SG suppressed the differentiation of 3T3-L1 cells stimulated by a mixture of 3-isobutyl-1-methylxanthine, dexamethasone, and insulin (MDI), and inhibited the proliferation of adipocytes during differentiation. Treatment of C. elegans with SG showed reductions in lipid accumulation by Nile red staining, thus directly demonstrating an antiobesity effect for SG. Furthermore, SG treatment down-regulated mRNA and protein expression levels of peroxisome proliferator-activated receptor subtype ${\gamma}$ ($PPAR{\gamma}$) and CCAAT/enhancer-binding protein-alpha ($C/EBP{\alpha}$) and decreased the mRNA level of sterol regulatory element-binding protein 1c in MDI-treated adipocytes in a dose-dependent manner. In differentiated 3T3-L1 cells, mRNA expression levels of lipid metabolism-regulating factors, such as amplifying mouse fatty acid-binding protein 2, leptin, lipoprotein lipase, fatty acid transporter protein 1, fatty acid synthase, and 3-hydroxy-3-methylglutaryl coenzyme A reductase, were increased, whereas that of the lipolytic enzyme carnitine palmitoyltransferase-1 was decreased. Our data demonstrate that SG inversely regulated the expression of these genes in differentiated adipocytes. SG induced increases in the mRNA expression of glycolytic enzymes such as glucokinase and pyruvate kinase, and a decrease in the mRNA level of the glycogenic enzyme phosphoenol pyruvate carboxylase. In addition, mRNA levels of the glucose transporters GLUT1, GLUT4, and insulin receptor substrate-1 were elevated by MDI stimulation, whereas SG dose-dependently inhibited the expression of these genes in differentiated adipocytes. SG also inhibited the phosphorylation of Akt (Ser473) at an early phase of MDI stimulation. Intracellular nitric oxide (NO) production and endothelial nitric oxide synthase mRNA levels were markedly decreased by MDI stimulation and recovered by SG treatment of adipocytes. Conclusion: Our results suggest that SG effectively inhibits adipocyte proliferation and differentiation through the downregulation of $PPAR{\gamma}$ and $C/EBP{\alpha}$, by suppressing Akt (Ser473) phosphorylation and enhancing NO production. These results provide strong evidence to support the development of SG for antiobesity treatment.