• Journal of Life Science
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• v.27 no.10
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• pp.1104-1110
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• 2017

SET1A is a histone H3K4 methyltransferase that catalyzes di- and trimethylation of histone H3 at lysine 4 (H3K4). Mono-, di-, and trimethylations on H3K4 (H3K4me1, H3K4me2, and H3K4me3, respectively) are generally correlated with gene activation. Although H3K4 methylation is associated with the stimulation of adipogenesis of 3T3-L1 preadipocytes, it remains unknown whether SET1A plays a role in the regulation of adipogenesis of 3T3-L1 preadipocytes. Here, we investigated whether SET1A regulates 3T3-L1 preadipocytes' adipogenesis and characterized the mechanism involved in this regulation. SET1A expression increased during 3T3-L1 preadipocytes' adipogenesis. Consistent with the increased SET1A expression, the global H3K4me3 level had also increased on day 2 after the induction of adipogenesis in 3T3-L1 adipocytes. SET1A knockdown using siRNA in 3T3-L1 preadipocytes inhibited 3T3-L1 preadipocytes' adipogenesis, as assessed by Oil Red O staining and the expression of adipogenic genes, indicating that SET1A stimulates the adipogenesis of 3T3-L1 preadipocytes. SET1A knockdown inhibited the cell proliferation of 3T3-L1 cells during mitotic clonal expansion (MCE) via down-regulation of the cell cycle gene cyclin E1, as well as the DNA synthesis gene, dihydrofolate reductase. Furthermore, SET1A knockdown repressed peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) expression during the late stage of adipogenesis. These results indicate that SET1A stimulates MCE and $PPAR{\gamma}$ expression, which leads to the promotion of 3T3-L1 preadipocytes' adipogenesis.

• Nutrition Research and Practice
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• v.15 no.3
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• pp.279-293
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• 2021

BACKGROUND/OBJECTIVES: The steamed ginger has been shown to have antioxidative effects and a protective effect against obesity. In the present study, we investigated the effects of ethanolic extract of steamed ginger (SGE) on adipogenesis in 3T3-L1 preadipocytes and diet-induced obesity (DIO) mouse model. MATERIALS/METHODS: The protective effects of SGE on adipogenesis were examined in 3T3-L1 adipocytes by measuring lipid accumulations and genes involved in adipogenesis. Male C57BL/6J mice were fed a normal diet (ND, 10% fat w/w), a high-fat diet (HFD, 60% fat w/w), and HFD supplemented with either 40 mg/kg or 80 mg/kg of SGE for 12 weeks. Serum chemistry was measured, and the expression of genes involved in lipid metabolism was determined in the adipose tissue. Histological analysis and micro-computed tomography were performed to identify lipid accumulations in epididymal fat pads. RESULTS: In 3T3-L1 cells, SGE significantly decreased lipid accumulation, with concomitant decreases in the expression of adipogenesis-related genes. SGE significantly attenuated the increase in body, liver, and epididymal adipose tissue weights by HFD. Serum total cholesterol and triglyceride levels were significantly lower in SGE fed groups compared to HFD. In adipose tissue, SGE significantly decreased adipocyte size than that of HFD and altered adipogenesis-related genes. CONCLUSIONS: In conclusion, steamed ginger exerted anti-obesity effects by regulating genes involved in adipogenesis and lipogenesis in 3T3-L1 cell and epididymal adipose tissue of DIO mice.

• KSBB Journal
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• v.27 no.6
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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 Physiology & Pathology in Korean Medicine
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• v.24 no.2
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• pp.266-271
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• 2010

Obesity occurred by energy imbalance, is increasing regardless of race, sex, age, and related to the metabolic syndrome, diabetes and cardiovascular disease. Since adipose tissue plays a critical role in regulating energy homeostasis, understanding of adipogenesis pathway and finding of regulatory mechanism for adipogenesis can be helpful to manage obesity as well as obesity-related diseases. In this study, to investigate the effects of Chestnut Inner Shell(CIS) extract on the adipogenesis in 3T3-L1 preadipocytes, 3T3-L1 preadipocytes were differentiated with adipogenic reagents for 9 days in the absence or presence of CIS extract ranging from 10 - 100 ${\mu}g/m{\ell}$. The effect of CIS extract on 3T3-L1 differentiation was examined by measuring intracelluar lipid droplet and triglyceride contents. CIS extract remarkably inhibited lipid accumulation(about 45% inhibition at 100 ${\mu}g/m{\ell}$ of CIS extract) and slightly decreased triglyceride contents(about 15% decrease at 100 ${\mu}g/m{\ell}$ of CIS extract) in 3T3-L1 preadipocytes at the concentration showing no cytotoxicity. These results demonstrated that CIS extract significantly inhibit adipogenesis and can be used for the regulation of obesity.

• The Korea Journal of Herbology
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• v.31 no.5
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• pp.47-53
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• 2016

Objectives : Mori Follium (Morus alba L. leaf) has been cultivated in many Asian countries. Especially, mulberry leaf has been used as an anti-diabetic remedy in oriental medicine. However, anti-obesity effect of mulberry has not been unknown. In this study, our objectives of study is to investigate the anti-adipogenic effect of mulberry water extract (MLE) and to reveal potential molecular anti-obesity mechanism in 3T3-L1 adipocytes differentiation model.Methods : The cytotoxicity of MLE in 3T3-L1 was examined by MTT assay. Anti-adipogenic effect of MLE was evaluated by Oil Red O (ORO) staining. To elucidate the molecular mechanism, inhibitor assay was employed. The mRNA expression levels of adipogenic transcriptional factors such as PPARγ and fatty acid synthase (FAS) were analyzed by reverse transcription-polymer chain reaction (RT-PCR) analysis.Results : The MLE treatment for 24 h did not affect to the 3T3-L1 cells at concentrations of 1, 10, 100, 200, 400, 800 and 1,000 ㎍/㎖. Thus, non-toxic concentration rages of MLE were used during adipogenesis period (day -2 to 7). Intracellular lipid accumulation in MLE-treated 3T3-L1 adipocytes (day 6) were quantitatively evaluated by ORO staining. The MLE treatment significantly and dose-dependently suppressed 3T3-L1 adipogenesis by 60.42%, 38.24%, and 5.97% at 10, 100, and 200 ㎍/㎖, respectively. In addition, our inhibitor assay and RT-PCR analysis revealed that the MLE-inhibited 3T3-L1 adipogenesis through inhibition of PPARγ mediated by Wnt/β-catenin signaling pathway.Conclusions : In conclusion, these findings indicate that the MLE could be used in prevent and/or treatment of obesity-related diseases.

• Korean Journal of Plant Resources
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• v.28 no.5
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• pp.582-590
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• 2015

In the present study, we investigated the effect of 70% EtOH extract from Hippophae Rhamnoides L. leaves (HRL) on the anti-obesity effect in 3T3-L1 cells. The effects of HRL on lipid accumulation in 3T3-L1 cells were examined using Oil Red O staining. In addition, we examined the gene expression levels by using RT-PCR and western blot. The results of this analysis showed that 100 ㎍/㎖ HRL significantly increased the inhibition of lipid accumulation by 82.25%; significantly decreased the mRNA expression of sterol regulatory element binding protein-1c (SREBP-1c), peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer binding proteins α (C/EBPα), and fatty acid synthase (FAS) in 3T3-L1 cells as well as the stimulated protein expression of AMP-activated protein kinase (AMPK); and suppressed the expression level of PPARγ. These results suggest that HRL can prevent adipogenesis through activation of AMPKα and inhibition of adipogenesis transcription factors.

• Preventive Nutrition and Food Science
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• v.22 no.2
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• pp.118-123
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• 2017

This study was conducted to investigate the anti-adipogenic activity of esculetin (ECT) which is reported to be attributable to the modulation of antioxidant enzymes during adipogenesis. After six days of ECT treatment of 3T3-L1 cells, lipid accumulation was determined by Oil red O staining. The levels of glutathione (GSH) and reactive oxygen species (ROS), and the activities of antioxidant enzymes including glutathione reductase, glutathione peroxidase (GPx), and catalase were examined. In addition, the protein expression of glutamate-cysteine ligase catalytic subunit (GCLC) and heme oxygenase-1 (HO-1) was measured by Western blot. ECT significantly inhibited lipid accumulation by approximately 80% and ROS production in a concentration-dependent manner. GSH level and GPx activity were increased by ECT by approximately 1.3-fold and 1.7-fold compared to the control group, respectively. GCLC and HO-1 expression were elevated by ECT. These results showed that ECT treatments strongly inhibit adipogenesis, increase GSH level, and upregulate the expression of GCLC and HO-1, possibly by decreasing ROS production in 3T3-L1 cells during adipogenesis.

• Journal of Physiology & Pathology in Korean Medicine
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• v.34 no.3
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• pp.136-141
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• 2020

The purpose of this study was to investigate the action mechanism of the roots of Adenophora triphylla var. japonica extract (ATE) in 3T3-L1 adipocytes. Cell toxicity test by MTT assay and lipid accumulation was performed to evaluate the inhibitory effect on the differentiation of adipocyte from preadipocytes induced by MDI differentiation medium, while adipogenesis related proteins expression level were evaluated by western blotting. As a result, ATE inhibited MDI-induced adipocyte differentiation in 3T3-L1 cells dose-dependently without cytotoxicity. Our results showed that ATE inhibited the phosphorylation of IRS1, thereby decreasing the expression of PI3K110α and reducing the phosphorylation of AKT and mTOR, resulting in attenuated protein expression of C/EBPα, PPARγ, ap2 and FAS in 3T3-L1 cells. These results suggest anti-adipogenic functions for ATE, and identified IRS1 as a novel target for ATE in adipogenesis.

• Nutrition Research and Practice
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• v.9 no.6
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• pp.592-598
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• 2015

BACKGROUND/OBJECTIVES: Increased mass of adipose tissue in obese persons is caused by excessive adipogenesis, which is elaborately controlled by an array of transcription factors. Inhibition of adipogenesis by diverse plant-derived substances has been explored. The aim of the current study was to examine the effects of the aqueous methanol extract of laver (Porphyra yezoensis) on adipogenesis and apoptosis in 3T3-L1 adipocytes and to investigate the mechanism underlying the effect of the laver extract. MATERIALS/METHODS: 3T3-L1 cells were treated with various concentrations of laver extract in differentiation medium. Lipid accumulation, expression of adipogenic proteins, including CCAAT enhancer-binding protein ${\alpha}$, peroxisome proliferator-activated receptor ${\gamma}$, fatty acid binding protein 4, and fatty acid synthase, cell viability, apoptosis, and the total content and the ratio of reduced to oxidized forms of glutathione (GSH/GSSG) were analyzed. RESULTS: Treatment with laver extract resulted in a significant decrease in lipid accumulation in 3T3-L1 adipocytes, which showed correlation with a reduction in expression of adipogenic proteins. Treatment with laver extract also resulted in a decrease in the viability of preadipocytes and an increase in the apoptosis of mature adipocytes. Treatment with laver extract led to exacerbated depletion of cellular glutathione and abolished the transient increase in GSH/GSSG ratio during adipogenesis in 3T3-L1 adipocytes. CONCLUSION: Results of our study demonstrated that treatment with the laver extract caused inhibition of adipogenesis, a decrease in proliferation of preadipocytes, and an increase in the apoptosis of mature adipocytes. It appears that these effects were caused by increasing oxidative stress, as demonstrated by the depletion and oxidation of the cellular glutathione pool in the extract-treated adipocytes. Our results suggest that a prooxidant role of laver extract is associated with its antiadipogenic and proapoptotic effects.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.6
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• pp.1409-1415
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• 2009

Obesity is especially a serious health problem in industrialized countries, because it is considered to be a risk factor associated with the genesis or development of various metabolic diseases, including cardiovascular disease and type 2 diabetes mellitus. The purpose of this study was to investigate the effects of tanshinone IIA from Salvia miltiorrhiza Bunge on induction of apoptossis and inhibition of adipogenesis in in 3T3-L1 preadipocytes and adipocytes. The results demonstrated that tanshinone IIA decreased cell population growth of 3T3-L1 preadipocytes, assessed with the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] and LDH (lactate dehydrogenase) assay. Flow cytometric analysis of 3T3-L1 preadipocytes exposed to tanshinone IIA showed that apoptotic cells increased in a timeand dose-dependent manner. Treatment with tanshinone IIA decreased the number of normal cells and increased the number of apoptotic cells in a dose-dependent manner. The induction of apoptosis in 3T3-L1 preadipocytes by tanshinone IIA was mediated through the activation of caspase-3 and Bax, and then through the cleavage of PARP and the down-regulation of Bcl-2. Moreover, tanshinone IIA significantly decreased the amount of intracellular triglycerides and GPDH (glycerol-3-phosphate dehydrogenase) activity in 3T3-L1 adipocytes. Our results suggest that tanshinone IIA efficiently induces apoptosis and inhibits adipogenesis in 3T3-L1 preadipocytes and adipocytes.