• Journal of Nutrition and Health
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• v.48 no.4
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• pp.327-334
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• 2015

Purpose: The objective of this study was to investigate the effects of (6)-gingerol, ginger components proliferation and adipocyte differentiation from early to lately steps. Methods: 3T3-L1 preadipocytes were cultured. Differentiation of confluent cells was induced with dexamethasone, isobutylxanthin and insulin for 2 day and cells were cultured by medium with insulin in presence of various concentrations 0, 25, 50, $100({\mu}mol/L)$ of (6)-gingerol for 4 day. Cell viability was measured using the EZ Cytox assay kit. In addition, we examined the expression of mRNA levels associated with each adipocyte differentiation step by real time reverse transcription polymerase chain reaction. Results: (6)-Gingerol inhibited adipocyte proliferation in a dose and time dependent manner. Expression of $C/EBP{\beta}$, associated with early differentiation step remained unchaged. However, intermmediate, late differentiation step and adipocytokines were effectively changed in dose-dependently manner in cell groups treated with (6)-gingerol. Conclusion: This study has shown that treatment with (6)-gingerol inhibited adipocyte proliferation as well as each adipocyte differentiation step. In particular, the (6)-gingerol more effectively inhibited adipocyte differentiation from intermmediate differentiation step.

• Journal of Life Science
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• v.32 no.7
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• pp.542-549
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• 2022

Polymethoxyflavones (PMFs) are flavonoids mainly found in citrus fruits and have been reported to exhibit a wide range of bioactivities, including anti-obesity, anti-cancer, and anti-inflammatory actions. To utilize PMFs as functional materials, it is necessary to develop a simple method of obtaining PMFs from citrus tissues containing a large amount of PMFs. It has been reported that Jinkyool (C. sunki Hort ex. Tanaka) peel contained a large amount of PMFs, but there are no studies on PMFs isolated from its leaves. In this study, we established a simple procedure for obtaining the PMF-rich fraction (PRF) from the leaves of Jinkyool and investigated the effects of PRF on lipid metabolism in 3T3-L1 cells. PRF inhibited lipogenesis during the differentiation of 3T3-L1 preadipocytes. It decreased the expression of peroxisome proliferator-activated receptor gamma (PPAR𝛾) and CCAAT/enhancer binding protein alpha (CEBP𝛼), FAS, and adipocyte fatty-acid-binding protein 2 (aP2). In mature 3T3-L1 adipocytes, PRF increases the phosphorylation of protein kinase A (PKA)/hormone-sensitive lipase (HSL), which are key factors involved in lipolysis. Moreover, it increases the phosphorylation of the AMP-activated protein kinase (AMPK)/acetyl-CoA carboxylase (ACC) involved in fatty acid oxidation. These results suggest that PRF from Jinkyool leaves can be used as an anti-obesity agent with the action of inhibiting lipogenesis and promoting lipolysis and fatty acid oxidation in 3T3-L1 adipocytes.

• BMB Reports
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• v.47 no.10
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• pp.587-592
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• 2014

We investigated the effects of ${\beta}$-adrenergic activation on bone marrow adiposity and on adipogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). C57BL/6 mice were subjected to a control (CON), high calorie (HIGH) or low calorie (LOW) diet for 12 weeks. In each group, mice were treated with vehicle (VEH) or propranolol. The number of adipocytes per area bone marrow was increased in LOWVEH and HIGHVEH mice compared with CONVEH mice, which was attenuated by propranolol. Isoproterenol increased lipid droplet accumulation and adipogenic marker gene expression in 3T3-L1 preadipocytes and mouse BMSCs, which were blocked by propranolol. Conditioned medium obtained from MC3T3-E1 osteoblasts suppressed adipogenic differentiation of 3T3-L1 cells, which was significantly attenuated by treatment of MC3T3-E1 cells with isoproterenol. These data suggest that ${\beta}$-adrenergic activation enhances bone marrow adipogenesis via direct stimulation of BMSCs adipogenesis and indirect inhibition of osteoblast anti-adipogenic potential.

• Journal of Life Science
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• v.25 no.3
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• pp.299-306
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• 2015

Chrysanthemum zawadskii, a herbaceous perennial plant belonging to the Compositae, grows wild in Asian countries, including Japan, China, and Korea. The biological, antioxidative, anti-inflammatory, and antibacterial activities of C. zawadskii have been reported, its antiobesity activity has not been elucidated. In the present study, the effect of C. zawadskii methanol extract (CZME) on pancreatic lipase enzyme activity, adipocyte differentiation, and adipogenesis was investigated using an in vitro assay and a cell model system. CZME effectively suppressed lipase enzyme activity in a dose-dependent manner. CZME also inhibited insulin, dexamethasone, 3-isobutyl-1-methylxanthine (MDI)-induced adipocyte differentiation, lipid accumulation, and the level of triglyceride in 3T3-L1 preadipocytes in a dose-dependent manner, without cytotoxicity. The antiobesity effect of CZME might be modulated by gene and protein expression of cytidine-cytidine-adenosine-adenosine-thymidine (CCAAT)/enhancer binding proteins (C/EBP) α, C/EBPβ, and the peroxisome proliferator-activated receptor γ (PPAR γ). CZME also triggered lipolysis in a dose-dependent manner in MDI-induced 3T3-L1 preadipocytes. Taken together, these results provide important new insights into the antiobesity activities of C. zawadskii, showing that they involve pancreatic lipase inhibition, as well as antiadipogenic and lipolysis effects. CZME might be a promising source in the field of nutraceuticals. However, the active compounds that confer the antiobesity activities of CZME need to be identified.

• Nutrition Research and Practice
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• v.5 no.1
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• pp.11-19
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• 2011

Eotaxin is an important inflammatory chemokine in eosinophil chemotaxis and activation and, thus, is implicated in asthma. Recently, obesity was associated with an increased prevalence of asthma, but the relationship between obesity and eotaxin expression has only been partially understood in obese mice and human studies. Therefore, we studied the expression patterns of eotaxin in 3T3-L1 preadipocytes/adipocytes to determine whether eotaxin levels are influenced by body weight gain and/or reduction in diet-induced obese mice. First, we investigated eotaxin expression during differentiation in 3T3-L1 adipocytes. Then, we treated 3T3-L1 preadipoeytes/adipoeytes with tumor necrosis factor-alpha (TNF-${\alpha}$), eotaxin, interleukin (IL)-4, IL-5, or leptin. To examine the effects of weight loss in high-fat diet induced obese mice, we fed C57BL/6 mice a high-fat diet or a normal diet for 26 weeks. Then, half of the high-fat diet group were fed a normal diet until 30 weeks to reduce weight. Epididymal adipose tissue, visceral adipose tissue, serum, and bronchoalveolar fluid of mice were examined for eotaxin expression. The results showed that eotaxin expression levels increased with adipocyte differentiation and that more eotaxin was expressed when the cells were stimulated with TNF-${\alpha}$, eotaxin, IL-4, IL-5, or leptin. An in vivo study showed that eotaxin levels were reduced in visceral adipose tissues when high-fat diet fed mice underwent weight loss. Taken together, these results indicate a close relationship between eotaxin expression and obesity as well as weight loss, thus, they indirectly show a relation to asthma.

• Journal of Sasang Constitutional Medicine
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• v.25 no.1
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• pp.51-61
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• 2013

Objectives We performed the simultaneous analysis for three compounds in Cheongsimyeonja-tang (CSYJT) and evaluated its anti-obesity effect. Methods The column for separation of three compounds was used Gemini $C_{18}$ column and maintained at $40^{\circ}C$. The mobile phase for gradient elution consisted of two solvent systems. The analysis was carried out at a flow rate of 1.0 mL/min with PDA detection at 275 nm. The injection volume was $10{\mu}L$. 3T3-L1 preadipocytes were differentiated into adipocytes by adding insulin, dexamethasone and 3-isobutyl-1-methylxanthine (IBMX) for 8 days in the absence or presence of CSYJT. Anti-obesity effects of CSYJT were evaluated by Oil Red O staining, glycerol-3-phosphate dehydrogenase (GPDH) activity, triglyceride contents, and leptin production. Results Calibration curves were acquired with $r^2$ >0.9999. The contents of baicalin, wogonoside and baicalein in CSYJT were 14.54-14.65 mg/g, 5.24-5.27 mg/g and 0.01-0.02 mg/g, respectively. CSYJT showed inhibitory effect on lipid accumulation and GPDH activity in the differentiated 3T3-L1 cells. Furthermore, CSYJT significantly decreased contents of triglyceride and leptin production in 3T3-L1 adipocytes. Conclusions These results will be helpful to improve quality control and anti-obesity effect of Taeeumin CSYJT.

• The Journal of Pediatrics of Korean Medicine
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• v.29 no.2
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• pp.37-48
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• 2015

Objectives This experimental study was designed to investigate the antiobese effects of Gamikwakhyangjungkisan and Fermented GamiKwakhyangjungkisan. Methods The cellular lipid contents were assessed by Oil-Red-O staining. The expression of $PPAR{\gamma}$ and $C/EBP{\alpha}$ were determined by real time RT-PCR and western blotting. In addition, body weight gain and serum lipid levels were measured in the mice with obesity induced by the high fat-diet for four weeks. Results Gamikwakhyangjungkisan and Fermented GamiKwakhyangjungkisan is reduced 3T3-L1 cells' differentiation and the expressions of $PPAR{\gamma}$ and $C/EBP{\alpha}$ in high concentration group. High-fat diet + Fermented GamiKwakhyangjungkisan group significantly reduced body weight gain. High-fat diet + Fermented GamiKwakhyangjungkisan group significantly increased HDL-cholesterol contents and reduced LDL-cholesterol contents. Furthermore, Fermented GamiKwakhyangjungkisan is excellent antiobese effects than Gamikwakhyangjungkisan. Conclusions These results demonstrate that Gamikwakhyangjungkisan and Fermented GamiKwakhyangjungkisan exerts antiobese effect in 3T3-L1 cells and mice fed high fat diet. Furthermore, Fermented GamiKwakhyangjungkisan is excellent antiobese effects than Gamikwakhyangjungkisan.

• Nutrition Research and Practice
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• v.7 no.3
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• pp.160-165
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• 2013

3T3-L1 preadipocyte were differentiated to adipocytes, and then treated with 0, 10, 20, and $40{\mu}g/mL$ of peanut sprout ethanol extract (PSEE). The main component of PSEE is resveratrol which contained 5.55 mg/mL of resveratrol. The MTT assay, Oil-Red O staining, glycerol-3-phosphate dehydrogenase (GPDH) activity, and the triglyceride concentration were determined in 3T3-L1 cells. MMP-2 and MMP-9 activities as well as mRNA expressions of C/EBP ${\beta}$ and C/EBP ${\alpha}$ were also investigated. As the concentration of PSEE in adipocytes increased, the cell proliferation was decreased in a dose-dependent manner from 4 days of incubation (P < 0.05). The GDPH activity (P < 0.05) and the triglyceride concentration (P < 0.05) were decreased as the PSEE treatment concentration increased. The mRNA expression of C/EBP${\beta}$ in 3T3-L1 cells was significantly low in groups of PSEE-treated, compared with control group (P < 0.05). The MMP-9 (P < 0.05) and MMP-2 (P < 0.05) activities were decreased in a dose-dependent manner as the PSEE concentration increased from $20{\mu}g/mL$. In conclusion, it was found that PSEE has an effect on restricting proliferation and differentiation of adipocytes.

• Nutrition Research and Practice
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• v.15 no.4
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• pp.444-455
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• 2021

BACKGROUND/OBJECTIVES: Adipocytes undergo angiogenesis to receive nutrients and oxygen needed for adipocyte' growth and differentiation. No study relating quercetin with angiogenesis in adipocytes exists. Therefore, this study investigated the role of quercetin on adipogenesis in 3T3-L1 cells, acting through matrix metalloproteinases (MMPs). MATERIALS/METHODS: After proliferating preadipocytes into adipocytes, various quercetin concentrations were added to adipocytes, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays were performed to evaluate cell proliferation. Glycerol-3-phosphate dehydrogenase (GPDH) activity was investigated as an indicator of fat accumulation. The mRNA expressions of transcription factors related to adipocyte differentiation, CCAAT/enhancer-binding proteins (C/EBPs), peroxisomal proliferatoractivated receptors (PPAR)-γ, and adipocyte protein 2 (aP2), were investigated. The mRNA expressions of proteins related to angiogenesis, vascular endothelial growth factor (VEGF)-α, vascular endothelial growth factor receptor (VEGFR)-2, MMP-2, and MMP-9, were investigated. Enzyme activities and concentrations of MMP-2 and MMP-9 were also measured. RESULTS: Quercetin treatment suppressed fat accumulation and the expressions of adipocyte differentiation-related genes (C/EBPα, C/EBPβ, PPAR-γ, and aP2) in a concentration-dependent manner in 3T3-L1 cells. Quercetin treatments reduced the mRNA expressions of VEGF-α, VEGFR-2, MMP-2, and MMP-9 in 3T3-L1 cells. The activities and concentrations of MMP-2 and MMP-9 were also decreased significantly as the concentration of quercetin increased. CONCLUSIONS: The results confirm that quercetin inhibits adipose tissue differentiation and fat accumulation in 3T3-L1 cells, which could occur through inhibition of the angiogenesis process related to MMPs.

• The Korea Journal of Herbology
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• v.29 no.4
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• pp.77-82
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• 2014

Objectives : The effects of ethanol extract of Plantago asiatica L. were investgated on adipocyte differentiation, lipopogenesis, lipolysis and apoptosis using differnentiated 3T3-L1 adipocytes. Methods : Plantago asiatica L. was extracted with ethanol (CCE). We carried on MTT assay for cell proliferation, Oil Red O staining for determination of cell differentiation and intracelluar adipogenesis. TUNEL staining assay for cell apoptosis, and Western blot analysis for measurement of pAMPK and pACC, $C/EBP{\alpha}$, $PPAR{\gamma}$ protein expressions were performed. Results : The addition of CCE up to 0.2 mg/ml into cell culture media showed no cytotoxicity. Treatment of 0.2 mg/ml CCE significantly inhibited differentiation in 3T3-L1 preadipocytes. Lipid accumulation of the CCE treated cells was decreased compared with that of control. Induction of cell apoptosis was increased in CCE treated cells compared with that of control. AMPK and ACC levels of the cells with 0.2 mg/ml CCE were led to phosphorylation and also expressions of $C/EBP{\alpha}$ and $PPAR{\gamma}$, as adipogenic transcription factors, were suppressed compared with those of control. Conclusions : Taken together, these results provide evidence that CCE has a regulatory role in lipid metabolism that is related to differentiation into adipocytes, adipogenesis and apoptosis.