• Title/Summary/Keyword: 3T3-L1 cells

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Pear pomace ethanol extract improves insulin resistance through enhancement of insulin signaling pathway without lipid accumulation

  • You, Mi-Kyoung;Kim, Hwa-Jin;Rhyu, Jin;Kim, Hyeon-A
    • Nutrition Research and Practice
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    • v.11 no.3
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    • pp.198-205
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    • 2017
  • BACKGROUND/OBJECTIVES: The anti-diabetic activity of pear through inhibition of ${\alpha}-glucosidase$ has been demonstrated. However, little has been reported about the effect of pear on insulin signaling pathway in obesity. The aims of this study are to establish pear pomace 50% ethanol extract (PPE)-induced improvement of insulin sensitivity and characterize its action mechanism in 3T3-L1 cells and high-fat diet (HFD)-fed C57BL/6 mice. MATERIALS/METHODS: Lipid accumulation, monocyte chemoattractant protein-1 (MCP-1) secretion and glucose uptake were measure in 3T3-L1 cells. Mice were fed HFD (60% kcal from fat) and orally ingested PPE once daily for 8 weeks and body weight, homeostasis model assessment of insulin resistance (HOMA-IR), and serum lipids were measured. The expression of proteins involved in insulin signaling pathway was evaluated by western blot assay in 3T3-L1 cells and adipose tissue of mice. RESULTS: In 3T3-L1 cells, without affecting cell viability and lipid accumulation, PPE inhibited MCP-1 secretion, improved glucose uptake, and increased protein expression of phosphorylated insulin receptor substrate 1 [p-IRS-1, ($Tyr^{632})$)], p-Akt, and glucose transporter type 4 (GLUT4). Additionally, in HFD-fed mice, PPE reduced body weight, HOMA-IR, and serum lipids including triglyceride and LDL-cholesterol. Furthermore, in adipose tissue, PPE up-regulated GLUT4 expression and expression ratio of p-IRS-1 ($Tyr^{632})/IRS$, whereas, down-regulated p-IRS-1 ($Ser^{307})/IRS$. CONCLUSIONS: Our results collectively show that PPE improves glucose uptake in 3T3-L1 cells and insulin sensitivity in mice fed a HFD through stimulation of the insulin signaling pathway. Furthermore, PPE-induced improvement of insulin sensitivity was not accompanied with lipid accumulation.

Hormonal Regulation of Leptin, Resistin, and Plasminogen Activator Inhibitor-1 Gene Expression in 3T3-L1 Adipocytes

  • Lee, Hyun-Jung;Kim, Yang-Ha
    • Preventive Nutrition and Food Science
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    • v.9 no.4
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    • pp.336-341
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    • 2004
  • Leptin, resisitn and PAI-1 (plasminogen activator inhibitor-1) are synthesized and secreted by rodent fat cells and recently postulated to be an important link to obesity. This study was conducted to characterize the hormonal regulation of leptin, resistin, and PAI-1 gene expression in the 3T3-L1 adipocytes. The cells were treated with 0.5 $\mu$M insulin, 1 $\mu$M dexamethasone (Dex), or 0.05 $\mu$M triiodothyronine (T3) for 72 hours. The mRNA levels of each peptide were measured by semi-quantitative RT-PCR. The mRNA level of the leptin-producing ob gene was significantly increased by insulin, Dex, and T3 by 3.2-, 3.1- and 2.7-fold, respectively, compared to the control (p < 0.05). The level of resistin mRNA was increased by insulin, Dex, and T3 by 2.7-, 2.5- and 2-fold, respectively, compared to the control (p < 0.05). Likewise, the level of PAI-1 mRNA was significantly increased by insulin, Dex, and T3 compared to the control (p < 0.05). Taken together, our results suggest that insulin, Dex, and T3 may regulate the gene expression of leptin, resistin, and PAI-1 in 3T3-L1 adipocytes.

Anti-adipogenic Effect of Chlorogenic Acid in 3T3-L1 Adipocytes

  • Park, Se-Eun;Choi, Jun-Hui;Lee, Hyo-Jeong;Seo, Kyoungsun;Kim, Seung
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.80-80
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    • 2018
  • Chlorogenic acid is a phenolic compound found in Cudrania tricuspidata fruits. In the present study, the effect of chlorogenic acid on the inhibition of adipogenesis in 3T3-L1 adipocytes was investigated. Cells were stained with Oil red O reagent to detect lipid droplets in adipocytes. The 3T3-L1 cells were lysed and measured for intracellular triglyceride and adipokine by ELISA kit. The protein expression of adipogenesis-related gene was evaluated by Western blot analysis. Chlorogenic suppressed lipid droplet and intracellular triglyceride accumulation in a concentration manner and also decreased secretion of adipokines such as leptin and adiponectin, compared with fully differentiated adipocytes. Treatment of 3T3-L1 cells with chlorogenic acid reduced the protein levels of peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) and, CCAAT/enhancer binding proteins alpha ($C/EBP{\alpha}$). This indicates that chlrogenic acid was effective as an anti-obesity agent by repressing the differentiation of 3T3-L1 into adipocytes and inhibiting triglyceridef formation in adipocyte and that it exerts its role mainly through the significant down-regulation of $PPAR{\gamma}$ and $C/EBP{\alpha}$.

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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.

Effects of Lonicera caerulea extract on adipocyte differentiation and adipogenesis in 3T3-L1 cells and mouse adipose-derived stem cells (MADSCs) (댕댕이나무 열매 추출물이 지방전구세포와 마우스 지방유래줄기세포의 분화 및 지방 생성 억제에 미치는 영향)

  • Park, Miey;Lee, Changho;Lee, Hae-Jeung
    • Journal of Nutrition and Health
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    • v.52 no.1
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    • pp.17-25
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    • 2019
  • Purpose: Obesity is a major health problem of global significance because it is clearly associated with an increased risk of health problems, such as nonalcoholic fatty liver disease (NAFLD), diabetes, cardiovascular diseases, and cancer. Lonicera caerulea (LC) originates from high mountains or wet areas and has been used as a traditional medicine in northern Russia, China, and Japan. LC contains a range of bioactive constituents, such as vitamins, minerals, and polyphenols. This study examined the anti-obesity effects of LC during differentiation in preadipocytes. Methods: The cell viability assay was performed after the differentiation of 3T3-L1 cells for 7 days. Oil Red O staining was used to visualize the changes in lipid droplets in 3T3-L1 cells and mouse adipose-derived stem cells (MADSCs). The mRNA expression of obesity-related genes was determined by quantitative real-time PCR. Results: According to the results of Oil Red O staining, the lipid levels and size of lipid droplets in the adipocytes were reduced and the LC extract (LCE, 0.25-1 mg/mL) markedly inhibited adipogenesis in a dose-dependent manner. The treatment of LCE also decreased the mRNA expression of peroxisome proliferator-activated receptor ${\gamma}$ ($PPAR{\gamma}$), CCAAT/enhancer binding protein-${\alpha}$ ($C/EBP{\alpha}$), and sterol regulatory element binding protein 1 (SREBP1) in 3T3-L1 cells. Western blot analysis showed that the $PPAR{\gamma}$, $C/EBP{\alpha}$, and SREBP1 protein levels in both 3T3-L1 and MADSC were reduced in a dose-dependent manner. Conclusion: These results suggest that LCE can inhibit adipogenic differentiation through the regulation of adipogenesis-related markers.

Crude Extract and Solvent-Partitioned Fractions of the Halophyte Atriplex gmelinii Inhibit Adipogenesis in 3T3-L1 Preadipocytes (3T3-L1 지방전구세포에서 염생식물 Atriplex gmelinii의 조추출물과 용매 분획물의 지방세포분화 억제)

  • Jung Im Lee;Jung Hwan Oh;Chang-Suk Kong;Youngwan Seo
    • Journal of Korean Medicine for Obesity Research
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    • v.23 no.2
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    • pp.69-77
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    • 2023
  • Objectives: Atriplex gmelinii C. A. Meyer is a halophyte belonging to the Chenopodiaceae family, and its young leaves and stems are used as fodder for livestock. The aim of the present study was to investigate the effects of A. gmelinii extract and its solvent fractions on lipid accumulation during adipogenesis of 3T3-L1 preadipocytes. Methods: The samples of A. gmelinii were separately extracted using methylene chloride and methanol. Subsequently, they were combined to formulate the initial extract, which was then partitioned based on polarity to prepare solvent fractions. Oil Red O staining was employed to measure lipid accumulation during the differentiation of 3T3-L1 preadipocytes. To verify cytotoxicity in 3T3-L1 cells, MTT assays were conducted. The expression levels of transcription factors in 3T3-L1 preadipocytes were measured through Western blotting analysis. Results: At 50 ㎍/mL, treatment of A. gmelinii extract and its solvent fractions during the differentiation of 3T3-L1 preadipocytes significantly diminished lipid accumulation with no noteworthy cytotoxicity on cell viability. Additionally, when investigating the biochemical pathways that underlie the prevention of lipid accumulation using solvent fractions, it was found that the n-BuOH and n-hexane fractions significantly decreased the expression of key transcription factors involved in the generation of fat, such as peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer binding protein α (C/EBPα), and sterol regulatory element-binding protein-1c (SREBP1c). Conclusions: These findings indicate that A. gmelinii can effectively reduce the accumulation of fat in 3T3-L1 adipocytes, making it a potentially valuable material for mitigating and preventing obesity.

Effects of the Extracts from Hoelen alba, Alismatis Rhizoma and Atractylodes Rhizoma on Proliferation and Differentiation of 3T3-L1 Cells (3T3-L1 세포(細胞)의 증식(增殖) 및 분화(分化)에 미치는 백복령(白茯笭), 택사(澤瀉) 및 창출(蒼朮)의 영향(影響))

  • Eun, Jae-Soon;Hong, Jong-Sung;So, June-No
    • Korean Journal of Pharmacognosy
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    • v.24 no.2
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    • pp.131-139
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    • 1993
  • These studies were conducted to investigate the effects of the extracts from Hoelen alba, Alismatis Rhizoma and Atractylodes Rhizoma on the proliferation and differentiation of 3T3-L1 cells. The results were summerized as follows: Hoelen alba and Alismatis Rhizoma extracts inhibited the proliferation of preadipose 3T3-L1 cells. In inductive differentiation, all three extracts inhibited the adipose conversion in 2 days of initial-culture, Atractylodes Rhizoma extract inhibited the adipose conversion in 5 days of final-culture and Hoelen alba and Alismatis Rhizoma inhibited adipose conversion in treatment of whole term of culture. In spontaneous differentiation, Atractylodes Rhizoma extract increased the adipose conversion in 2 days of initial-culture, Hoelen alba and Alismatis Rhizoma increased the adipose conversion in 5 days of final-culture, all three extracts increased adipose conversion in treatment of whole term of culture. The 10% serum of mice treated with each sample did not affect, but the 5% serum of them inhibited the proliferation of 3T3-L1 cells. In inductive differentiation, the 10% serum of them inhibited the adipose conversion in treatment of whole term of culture.

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Antiadipogenic Effects of Salvia plebeia R. Br. Extracts by Extraction Conditions in 3T3-L1 Preadipocytes (추출조건에 따른 배암차즈기 추출물의 지방세포 분화 및 지방축적 억제 효과)

  • Choi, Soo Im;Kwak, Hoyoung;Kim, Jae Yoon;Choi, Jong Gil;Lee, Je Hyuk
    • Korean Journal of Medicinal Crop Science
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    • v.23 no.3
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    • pp.245-252
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    • 2015
  • This study was carried out to investigate the effects of Salvia plebeia R. Br. ethanolic extract with different aspects (stem/leaf and whole plant) on differentiation and lipid accumulation in 3T3-L1 preadipocytes. The morphological changes and the degrees of lipid accumulation in 3T3-L1 cells were measured by Oil Red O staining and intra-cellular triglyceride (TG) assay. The mRNA expressions of special peroxisome proliferation activated receptor- genes (PPAR), CCAAT/ enhancer-binding protein (C/$EBP{\alpha}$), fatty acid synthase (FAS) and lipoprotein lipase (LPL) were detected by reverse transcriptase polymerase chain reaction (RT-PCR). The 50% ethanolic extracts ($100{\mu}g/mL$) of stem and leaf (SALE) and 30% ethanolic extracts (100 g/mL) of whole plant (SAE) from Salvia plebeia R. Br. were significantly attenuated lipid accumulation during adipogenesis in 3T3-L1 cells. Ethyl acetate-soluble fractions ($50{\mu}g/mL$) significantly inhibited lipid droplet accumulation in 3T3-L1 cells. In addition, SALE induced down-regulation of specific adipogenic transcriptional factors (C/$EBP{\alpha}$ and $PPAR{\gamma}$) and target genes (FAS and LPL) during adipogenesis. Salvia plebeia R. Br. may be used as a safe and efficient natural substance to manage obesity.

Convergence study on the through inhibition of differentiation in 3T3-L1 cells of ethanol extract from Trichosanthes kirilowii Maxim. Root (하늘타리(Trichosanthes kirilowii Maxim.) 뿌리 에탄올 추출물의 3T3-L1 지방세포 분화 억제 융합연구)

  • Kim, Sung Ok;Jeung, Ji-Suk
    • Journal of the Korea Convergence Society
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    • v.10 no.3
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    • pp.127-133
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    • 2019
  • The ami of our study was on the anti-obesity effect of ethanol extract from Trichosanthes kirilowii Maxim root (TKM) in murine adipocytes, 3T3-L1 cells. This study focused on anti-adipogenic activity through inhibition of cell differentiation in 3T3-L1 cells treated TKM. 100 ug/ml of non-cytotoxic TEM remarkablely inhibited content of triglycerol and suppressed expressions of $C/EBP{\alpha}$, $PPAR{\gamma}a$ and SREBP-1c related with lipogenic transcription factors in theres 3T3-L1 cells compared to (-)control cells. As phosphorylations of AMPK and ACC were incerased, HSL and CPT-1 mRNA expression increased upon TKM treatment, which involved in inhibition of fatty acid synthase expression. In conclusion, these results indicate that TKM can inhibit mRNA and protein expression of lipogenic genes in 3T3-L1 adipocytes. Our study suggests that TKM has potential anti-obesity effects and is a convergence therapeutic functional agent with anti-adipogenic activity via hypolipogenesis.

Effect of Water Extract of Carthamus tinctorious L. on In Vitro Activity of T and B Lymphocytes (시험관내에서 홍화의 물 추출물이 T 및 B 림프구의 활성에 미치는 영향)

  • Choi, Youn-Hwa;Do, Jeong-Su;Nam, Sang-Yun
    • Korean Journal of Pharmacognosy
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    • v.35 no.4 s.139
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    • pp.330-337
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    • 2004
  • Based on the traditional application of Carthamus tinctorious L. (CF) as a component of Korean medicinal decoctions, in the present study, we investigated in vitro an immunomodulatory activity of water extract of CF(WECF). Water extract of CF significantly increased the in vitro proliferative responses of spleen cells (SPC). However, addition of WECF during anti-CD3 activation resulted in a significant decrease in SPC proliferation. Flow cytometric analysis showed that WECF addition chanced T and B cell frequencies in anti-CD3-activated spleen cell populations. Using purified cells, it was revealed that WECF is mitogenic to B cells but rather inhibitory to T cell Proliferation. Upon anti-CD3 stimulation, high concentration (1 mg/ml) of WECF significantly inhibited T cell proliferation until day 2 of stimulation. At day 3, anti-CD3-activated cells exposed to WECF recovered their proliferation to the level comparable to control. Although B cell proliferation was also inhibited in proliferation at day 1, it recovered sooner and then was rather augmented by WECF at day 3. These data indicate that WECF down-regulates lymphocyte proliferation at early phase of activation but T cells are more vulnerable than B cells to WECF, However, CD4+ and CD8+ T cells did not differ in WECF-mediated immunotoxicity. Data of propidium iodide (PI) staining showed that WECF accelerates activated T cell, but not B cell, apoptosis and WECF concurrently inhibited cytokine production of activated T cells. Taken together, WECF exhibits B cell mitogenic activity and differential toxicity more pronounced to T cells, suggesting a possible in vivo application of WECF for specific control of T cells without alteration of B cell activity.