• Title/Summary/Keyword: 3T3-L1 adipocytes

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Korean Curcuma longa L. induces lipolysis and regulates leptin in adipocyte cells and rats

  • Song, Won-Yeong;Choi, Jeong-Hwa
    • Nutrition Research and Practice
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    • v.10 no.5
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    • pp.487-493
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    • 2016
  • BACKGROUND/OBJECTIVES: Turmeric (Curcuma longa L.) has been reported to have many biological functions including anti-obesity. Leptin, peptide hormone produced by adipocytes and its concentration is increased in proportion to the amount of the adipocytes. In the present study, we examined the effects of Korean turmeric on the regulation of adiposity and leptin levels in 3T3-L1 adipocytes and rats fed a high-fat and high-cholesterol diet. MATERIALS/METHODS: Leptin secretion, free fatty acid and glycerol contents in 3T3-L1 adipocytes were measured after incubation of cells with turmeric for 24 hours. Rats were divided into four experimental groups: a normal diet group (N), a high-fat and high-cholesterol diet group (HF), a high-fat and high-cholesterol diet group supplemented with 2.5% turmeric extracts (TPA group) and a high-fat and high-cholesterol diet group supplemented with 5% turmeric extracts (TPB group). Serum samples were used for the measurement of leptin concentration. RESULTS: Contents of free fatty acid and glycerol showed concentration dependent increase in response to turmeric extracts. Effects of turmeric extracts on reduction of lipid accumulation in 3T3-L1 cells were examined by Oil Red O staining. Treatment with turmeric extracts resulted in increased expression levels of adipose triglyceride lipase and hormone-sensitive lipase mRNA. The concentration of leptin from 3T3-L1 adipocytes was significantly decreased by turmeric. Proportional abdominal and epididymal fats weights of the turmeric 5% supplemented group, TPB has significantly decreased compared to the HF group. The serum levels of leptin in the TPA and TPB groups were significantly lower than those of the HF group. CONCLUSIONS: Based on these results, we suggested that Korean turmeric may contribute to the decreasing of body fat and regulating leptin secretion.

Anti-adipogenic Effect of Undaria pinnatifida Extracts by Ethanol in 3T3-L1 Adipocytes (미역 에탄올 추출물이 지방세포 형성과정에 미치는 영향)

  • Kim, Hye-Jin;Kang, Chang-Han;Kim, Sung-Koo
    • 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.

Anti-adipogenic effect of mulberry leaf ethanol extract in 3T3-L1 adipocytes

  • Yang, Soo Jin;Park, Na-Young;Lim, Yunsook
    • Nutrition Research and Practice
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    • v.8 no.6
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    • pp.613-617
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    • 2014
  • BACKGROUND/OBJECTIVES: Adipogenesis is part of the cell differentiation process in which undifferentiated fibroblasts (pre-adipocytes) become mature adipocytes with the accumulation of lipid droplets and subsequent cell morphological changes. Several transcription factors and food components have been suggested to be involved in adipogenesis. The aim of this study was to determine whether mulberry leaf ethanol extract (MLEE) affects adipogenesis in 3T3-L1 adipocytes. MATERIALS/METHODS: The 3T3-L1 adipocytes were treated with different doses of MLEE for 8 days starting 2 days post-confluence. Cell viability, fat accumulation, and adipogenesis-related factors including CCAAT-enhancer-binding protein alpha ($C/EBP{\alpha}$), peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$), $PPAR{\gamma}$ coactivator 1 alpha (PGC-$1{\alpha}$), fatty acid synthase (FAS), and adiponectin were analyzed. RESULTS: Results showed that MLEE treatments at 10, 25, 50, and $100{\mu}g/ml$ had no effect on cell morphology and viability. Without evident toxicity, all MLEE treated cells had lower fat accumulation compared with control as shown by lower absorbances of Oil Red O stain. MLEE at 50 and $100{\mu}g/ml$ significantly reduced protein levels of $PPAR{\gamma}$, PGC-$1{\alpha}$, FAS, and adiponectin in differentiated adipocytes. Furthermore, protein level of $C/EBP{\alpha}$ was significantly decreased by the treatment of $100{\mu}g/ml$ MLEE. CONCLUSION: These results demonstrate that MLEE treatment has an anti-adipogenic effect in differentiated adipocytes without toxicity, suggesting its potential as an anti-obesity therapeutic.

Green Tea (-)-Epigallotocatechin-3-Gallate Induces PGC-1α Gene Expression in HepG2 Cells and 3T3-L1 Adipocytes

  • Lee, Mak-Soon;Lee, Seohyun;Doo, Miae;Kim, Yangha
    • Preventive Nutrition and Food Science
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    • v.21 no.1
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    • pp.62-67
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    • 2016
  • Green tea (Camellia sinensis) is one of the most popular beverages in the world and has been acknowledged for centuries as having significant health benefits. (-)-Epigallocatechin-3-gallate (EGCG) is the most abundant catechin in green tea, and it has been reported to have health benefit effects. Peroxisome proliferator-activated receptor ${\gamma}$ coactivator $(PGC)-1{\alpha}$ is a crucial regulator of mitochondrial biogenesis and hepatic gluconeogenesis. The objective of this study was to investigate whether EGCG from green tea can affect the ability of transcriptional regulation on $PGC-1{\alpha}$ mRNA expression in HepG2 cells and 3T3-L1 adipocytes. To study the molecular mechanism that allows EGCG to control $PGC-1{\alpha}$ expression, the promoter activity levels of $PGC-1{\alpha}$ were examined. The $PGC-1{\alpha}$ mRNA level was measured using quantitative real-time PCR. The -970/+412 bp of $PGC-1{\alpha}$ promoter was subcloned into the pGL3-Basic vector that includes luciferase as a reporter gene. EGCG was found to up-regulate the $PGC-1{\alpha}$ mRNA levels significantly with $10{\mu}mol/L$ of EGCG in HepG2 cells and differentiated 3T3-L1 adipocytes. $PGC-1{\alpha}$ promoter activity was also increased by treatment with $10{\mu}mol/L$ of EGCG in both cells. These results suggest that EGCG may induce $PGC-1{\alpha}$ gene expression, potentially through promoter activation.

Intensive Proteomic Approach to Identify Secreted Peptides/Proteins from 3T3-L1 Adipocytes using Gel Electrophoresis and Liquid Chromatograph Separation Methods (젤 전기영동 및 액체 크로마토그래피 분리 방법을 이용하여 지방 세포로부터 분비되는 단백질들에 대한 프로테오믹스 연구 방법)

  • Hwang, Hyun-Ho;Baek, Moon-Chang
    • YAKHAK HOEJI
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    • v.55 no.3
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    • pp.203-212
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    • 2011
  • Adipocytes have been known to secrete a number of important proteins called adipokines with roles in energy metabolism, reproduction, cardiovascular function and immunity. In this study we have attempted to identify intensively secretory proteins from 3T3-L1 adipocytes. 3T3-L1 preadipocytes were differentiated into mature adipocytes and then the cells were left in serum-free medium. The supernatant was filtrated and dialyzed. Lyophilized secretome was fractionated by two different methods, 1-D SDS PAGE and RP-FPLC. The tryptic peptides from the gel slices and the FPLC fractions were analyzed by nanoLC/ESI-MS/MS. We identified a total of 303 identical proteins from two methods, 251 proteins from 1-D gel and 184 proteins from RP-FPLC. 86 of them were listed as a secretory protein Finally, we identified many known or unknown secreted proteins existed in the low level including adiponectin, angiotensinogen, bone morphogenetic protein-1 (BMP-1), macrophage migration inhibitory factor (MIF), insulin like growth factor-II (IGF-II), interleukin-6 (IL-6), follistatin-related protein-1, minecan, and resistin. The existence of some of secreted proteins has been confirmed in RNA level. This proteomic experiment is useful for the intensive screening of secretory proteins in many kinds of other cells.

Effect of Eriobotrya japonica Leaf and Seed Extracts on Adipogenesis (비파잎과 씨 추출물의 지방생성 억제효과)

  • Min, Oh-Jin;Oh, Jin;Kim, Hyeon-A;Kim, Min-Sook;Baek, Hum-Young;Kim, Yong-Jae;Rhyu, Dong-Young
    • Korean Journal of Pharmacognosy
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    • v.41 no.4
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    • pp.270-274
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    • 2010
  • Obesity is caused from an imbalance between energy intake and expenditure, which may lead to pathologic growth of adipocytes and accumulation of fat in tissue. We examined the inhibitory effects of Eriobotrya japonica leaf and seed extracts on lipid absorption in vitro and fat accumulation during the differentiation of 3T3-L1 to adipocytes. 3T3-L1 preadipocytes were stimulated with DMEM media containing 10% FBS, 0.5 mM 3-isobuthyl-1-methyxanthine (IBMX), $5\;{\mu}g/ml$ insulin, and $1\;{\mu}g/ml$ dexamethasone for differentiation to adipocytes. E. japonica leaf extract at concentration of 0.5 or 1 mg/ml inhibited pancreatic lipase activity. The cell viability of 3T3-L1 adipocytes slightly reduced about 3% by treatment of E. Japonica leaf and seed extracts. The leaf and seed extracts of E. japonica effectively inhibited the accumulations of lipid droplet and expression of $C/EBP{\alpha}$ promoting adipogenesis. Thus, this data suggest that E. japonica leaf and seed extracts inhibit fat accumulation through regulation of $C/EBP{\alpha}$, and leaf extract is more effective in lipid absorption and adipogenesis than seed extract.

Novel Role of Dipterocarpus tuberculatus Roxb. as a Lipogenesis Inhibitor and Lipolysis Stimulator in 3T3-L1 Adipocytes (3T3-L1 지방세포에서 lipogenesis 저해제와 lipolysis 촉진제로서 Dipterocarpus tuberculatus Roxb.의 새로운 역할)

  • Su Jin, Lee;Ji Eun, Kim;Yun Ju, Choi;You Jeong, Jin;Yu Jeong, Roh;AYun, Seol;Hee Jin, Song;Dae Youn, Hwang
    • Journal of Life Science
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    • v.32 no.11
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    • pp.855-864
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    • 2022
  • The pharmacological efficacy of Dipterocarpus tuberculatus Roxb. has been verified in only several fields including photoaging, inflammation, hepatotoxicity, acute gastritis and osseointegration. To identify the novel functions of Dipterocarpus tuberculatus Roxb. on anti-obesity, inhibitory effect on lipid accumulation and stimulatory effect on lipolysis were investigated in MDI (3-isobutyl-1-methyl-xanthine, dexamethasone, and insulin) stimulated 3T3-L1 adipocytes treated with methanol extracts of Dipterocarpus tuberculatus Roxb. (MED). Lipogenic targets, including lipid accumulation, level of lipogenic transcription factors, and expression of lipogenic regulators, were downregulated in MDI-stimulated 3T3-L1 adipocytes treated with MED without any significant cytotoxicity. Also, MED treatment inhibited the mRNA levels of adipogenic targets including peroxisome proliferator-activated receptor (PPAR)γ and CCAAT-enhancer binding protein (C/EBP) α, as well as lipogeic targets including adipocyte fatty acid binding protein 2 (aP2) and fatty acid synthetase (FAS) in MDI-stimulated 3T3-L1 adipocytes. A similar decrease patterns were detected in Oil red O stained lipid droplets of MED treated MDI-stimulated 3T3-L1 adipocytes. Furthermore, several lipolytic targets, such as cAMP concentration, concentration of free glycerol, expression level of lipases, including ATGL, perilipin and HSL, were upregulated in MDI-stimulated 3T3-L1 adipocytes treated with MED. These results show that MED has a novel role as a lipogenesis inhibitor and lipolysis stimulator in MDI-stimulated 3T3-L1 adipocytes.

Expression of eotaxin in 3T3-L1 adipocytes and the effects of weight loss in high-fat diet induced obese mice

  • Kim, Hyun-Jung;Kim, Chang-Hyun;Lee, Do-Hyun;Han, Min-Woo;Kim, Mi-Young;Ju, Jae-Hyun;Do, Myoung-Sool
    • 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.

Effects of Yeoldahanso-tang, a Sasang Constitutional Herbal Prescription for Taeeumin, on the Adipogenesis in 3T3-L1 Cells (태음인(太陰人) 열다한소탕(熱多寒少湯)이 3T3-L1 지방전구세포(脂肪前驅細胞) 분화(分化)에 미치는 영향(影響))

  • Yoo, Sae-Rom;Jeong, Soo-Jin;Shin, Hyeun-Kyoo
    • Journal of Sasang Constitutional Medicine
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    • v.24 no.4
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    • pp.75-83
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    • 2012
  • Objectives : Although Yeoldahanso-tang (YDHST) has been widely used for treatment of obesity and its related diseases such as hyperinsulinemia and hypertension for Taeeumin, no scientific evidence has reported yet to support its ability to work against these metabolic disorders. Our study was aimed to investigate the anti-obesity effect of YDHST extract on the cellular differentiation of 3T3-L1 preadipocytes into adipocytes. Methods : 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 YDHST extract. Anti-obesity effects of YDHST extract were evaluated by Oil Red O staining, glycerol-3-phosphate dehydrogenase (GPDH) activity, triglyceride contents, and leptin production. Results : YDHST extract remarkably prevented lipid accumulation with no cytotoxicity in the differentiated 3T3-L1 cells. In addition, YDHST extract decreased contents of triglyceride 3T3-L1 adipocytes. Consistently, YDHST extract caused a significant inhibition of GPDH activity and leptin production in a dose-dependent manner. Conclusions : Our findings suggest that Sasang constitutional herbal formula YDHST for Tae-eumin has anti-obesity activity by regulation of the adipogenesis process in vitro. Additional study will be required to further confirm the inhibitory effect on adipocyte differentiation by using in vivo animal model.

Peanut sprout ethanol extract inhibits the adipocyte proliferation, differentiation, and matrix metalloproteinases activities in mouse fibroblast 3T3-L1 preadipocytes

  • Kim, Woo Kyoung;Kang, Nam E;Kim, Myung Hwan;Ha, Ae Wha
    • 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.