• Title, Summary, Keyword: adipocyte differentiation

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Homeostatic balance of histone acetylation and deconstruction of repressive chromatin marker H3K9me3 during adipocyte differentiation of 3T3-L1 cells

  • Na, Han?Heom;Kim, Keun?Cheol
    • Genes and Genomics
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    • v.40 no.12
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    • pp.1301-1308
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    • 2018
  • Background Adipocyte differentiation is completed by changing gene expression. Chromatin is closely related to gene expression. Therefore, its structure might be changed for adipocyte differentiation. Mouse 3T3-L1 preadipocytes have been used as a cell model to study molecular mechanisms of adipogenesis. Objective To examine changes of chromatin modification and expression of histone modifying enzymes during adipocyte differentiation. Methods Microscopic analysis and Oil Red O staining were performed to determine distinct phenotype of adipocyte differentiation. RT-PCR and Western blot analysis were used to examine expression levels of histone modifying enzymes during adipocyte differentiation. Histone modifications were examined by immunostaining analysis. Results Expression levels of P300 and cbp were increased during adipocyte differentiation. However, acetylation of histones was not quantitatively changed postdifferentiation of 3T3-L1 cells compared to that at pre-differentiation. RT-PCR and Western blot analyses showed that expression levels of hdac2 and hdac3 were increased during adipocyte differentiation, suggesting histone acetylation at chromatin level was homeostatically controlled by increased expression of both HATs and HDACs. Tri-methylation level of H3K9 (H3K9me3), but not that of H3K27me3, was significantly decreased during adipocyte differentiation. Decreased expression of setdb1 was consistent with reduced pattern of H3K9me3. Knock-down of setdb1 induced adipocyte differentiation. This suggests that setdb1 is a key chromatin modifier that modulates repressive chromatin. Conclusion These results suggest that there exist extensive mechanisms of chromatin modifications for homeostatic balance of chromatin acetylation and deconstruction of repressive chromatin during adipocyte differentiation.

Roles of Protein Histidine Phosphatase 1 (PHPT1) in Brown Adipocyte Differentiation

  • Kang, Joo Ae;Kang, Hyun Sup;Bae, Kwang-Hee;Lee, Sang Chul;Oh, Kyoung-Jin;Kim, Won Kon
    • Journal of Microbiology and Biotechnology
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    • v.30 no.2
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    • pp.306-312
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    • 2020
  • Despite the importance of brown adipocytes as a therapeutic target for the prevention and treatment of obesity, the molecular mechanism underlying brown adipocyte differentiation is not fully understood. In particular, the role of post-translational modifications in brown adipocyte differentiation has not been extensively studied. Histidine phosphorylation is increasingly recognized an important process for protein post-translational modifications. In this study, we show that histidine phosphorylation patterns change during brown adipocyte differentiation. In addition, the expression level of protein histidine phosphatase 1 (PHPT1), a major mammalian phosphohistidine phosphatase, is reduced rapidly at the early phase of differentiation and recovers at the later phase. During white adipocyte differentiation of 3T3-L1 preadipocytes, however, the expression level of PHPT1 do not significantly change. Knockdown of PHPT1 promotes brown adipocyte differentiation, whereas ectopic expression of PHPT1 suppresses brown adipocyte differentiation. These results collectively suggest that histidine phosphorylation is closely linked to brown adipocyte differentiation and could be a therapeutic target for obesity and related metabolic diseases.

Methyltransferase and demethylase profiling studies during brown adipocyte differentiation

  • Son, Min Jeong;Kim, Won Kon;Oh, Kyoung-Jin;Park, Anna;Lee, Da Som;Han, Baek Soo;Lee, Sang Chul;Bae, Kwang-Hee
    • BMB Reports
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    • v.49 no.7
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    • pp.388-393
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    • 2016
  • Although brown adipose tissue is important with regard to energy balance, the molecular mechanism of brown adipocyte differentiation has not been extensively studied. Specifically, regulation factors at the level of protein modification are largely unknown. In this study, we examine the changes in the expression level of enzymes which are involved in protein lysine methylation during brown adipocyte differentiation. Several enzymes, in this case SUV420H2, PRDM9, MLL3 and JHDM1D, were found to be up-regulated. On the other hand, Set7/9 was significantly down-regulated. In the case of SUV420H2, the expression level increased sharply during brown adipocyte differentiation, whereas the expression of SUV420H2 was marginally enhanced during the white adipocyte differentiation. The knock-down of SUV420H2 caused the suppression of brown adipocyte differentiation, as compared to a scrambled control. These results suggest that SUV420H2, a methyltransferase, is involved in brown adipocyte differentiation, and that the methylation of protein lysine is important in brown adipocyte differentiation.

Inhibitory Effects of Cultivated Wild Ginseng on the Differentiation of 3T3-L1 Pre-adipocytes

  • Mollah, Mohammad Lalmoddin;Cheon, Yong-Pil;In, Jun-Gyo;Yang, Deok-Chun;Kim, Young-Chul;Song, Jae-Chan;Kim, Kil-Soo
    • Journal of Ginseng Research
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    • v.35 no.1
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    • pp.45-51
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    • 2011
  • Wild ginseng has been used as a traditional medicine for thousands of years and for increase physical strength in Korea, China and Japan. This study reports that cultivated wild ginseng (CWG) inhibits adipocyte differentiation of 3T3-L1 pre-adipocytes in a concentration-dependent manner. Inhibition of adipocyte differentiation is one possible anti-obesity strategy. CWG inhibits the expression of the adipocyte differentiation regulator peroxisome proliferators-activated receptor (PPAR)${\gamma}$ and CCAAT/enhancer-binding protein ${\alpha}$mRNA. It also inhibited the expression of PPAR${\gamma}$ and adiponectin at the protein level during the differentiation of pre-adipocytes into adipocytes. Additionally, CWG blocked the cell cycle at the sub-$G_1$ phase transition, causing cells to remain in the pre-adipocyte state. These results indicate that CWG inhibits adipocyte differentiation and adipogenesis through pre-adipocyte cell cycle arrest in cultured 3T3-L1 cells.

15-Hydroxyprostaglandin Dehydrogenase Is Associated with the Troglitazone-Induced Promotion of Adipocyte Differentiation in Human Bone Marrow Mesenchymal Stem Cells

  • Noh, Min-Soo;Lee, Soo-Hwan
    • Biomolecules & Therapeutics
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    • v.18 no.1
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    • pp.16-23
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    • 2010
  • Adipocyte differentiation in human bone marrow mesenchymal stem cells (hBM-MSCs) is not as efficient as that in murine pre-adipocytes when induced by adipogenic agents including insulin, dexamethasone, and 3-isobutyl-1-methylxanthine (IDX condition). Therefore, the promotion of adipocyte differentiation in hBM-MSCs has been used as a cell culture model to evaluate insulin sensitivity for anti-diabetic drugs. In hBM-MSCs, $PPAR{\gamma}$ agonists or sulfonylurea anti-diabetic drugs have been added to IDX conditions to promote adipocyte differentiation. Here we show that troglitazone, a peroxisome proliferator-activated receptor-gamma ($PPAR{\gamma}$) agonist, significantly reduced the levels of anti-adipogenic $PGE_2$ in IDX-conditioned hBM-MSC culture supernatants when compared to $PGE_2$ levels in the absence of $PPAR{\gamma}$ agonist. However, there was no difference in the mRNA levels of cyclooxygenases (COXs) and the activities of COXs and prostaglandin synthases during adipocyte differentiation in hBM-MSCs with or without troglitazone. In hBM-MSCs, troglitazone significantly increased the mRNA level of 15-hydroxyprostaglandin dehydrogenase (HPGD) which can act to decrease $PGE_2$ levels in culture. These results suggest that the role of $PPAR{\gamma}$ activation in promoting adipocyte differentiation in hBM-MSCs is to reduce anti-adipogenic $PGE_2$ levels through the up-regulation of HPGD expression.

Effects of (6)-gingerol, ginger component on adipocyte development and differentiation in 3T3-L1 (생강 성분인 (6)-Gingerol이 3T3-L1에서 지방세포 증식과 분화 과정에 미치는 영향)

  • Seo, Eun Young
    • 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.

Evaluation of the inhibition of the differentiation of pre-adipocytes into matures adipocytes

  • Morvan, Pierre Yves
    • Proceedings of the SCSK Conference
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    • pp.440-447
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    • 2003
  • Up until today, the key to contouring has been resumed in these two alternatives, either limiting the adipocyte storing capacity by modulating lipogenesis, or by stimulating lipolysis to eliminate adipocyte lipid content. Another interesting way could be the regulation of adipocyte differentiation. In this work, we have evaluated the effect of a brown algal extract of Sphacelaria scoparia (SSE) on the differentiation of pre-adipocytes into adipocytes. A pre-adipocyte line (3T3-L 1) was used. The differentiation was evaluated by the measure of produced lipids thanks to red oil coloration and spectrophotometry, and also by the expression of adipocyte differentiation markers: enzymes such as fatty acid synthase (FAS) and stearoyl CoA desaturase (SCD), or membrane proteins such as glucose transporters (GLUT -4) and fatty acid transporters (FAT) expressed on the surface of human adipocytes. These genes are under control of two transcription factors: CAAT-enhancer binding protein (c/EBP alpha) and sterol response element binding protein (SREBP1). All these markers were analysed at different stages of differentiation by RT -PCR. Sphacelaria extract (SSE) inhibits pre-adipocytes differentiating into adipocytes following a dose-dependant relation, using a kinetics similar to retinoic acid. It decreases the expression of mRNA specific to FAS, FAT, GLUT -4, SCD1, c/EBP alpha and SREBP1. Moreover, SSE regulated on collagen 1 and collagen 4 expression. A stimulation of collagen 1 was also measured in human skin fibroblasts. Thus, SSE performs as a genuine differentiation inhibitor and not only as a lipogenesis inhibitor, and could be used in slimming products.

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TonEBP suppresses adipocyte differentiation via modulation of early signaling in 3T3-L1 cells

  • Kim, Soo Jin;Kim, Taehee;Choi, Han Na;Cho, Eun Jung;Park, Jin Bong;Jeon, Byeong Hwa;Lee, Sang Do
    • The Korean Journal of Physiology and Pharmacology
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    • v.20 no.6
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    • pp.649-655
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    • 2016
  • TonEBP belongs to the Rel family of transcription factors and plays important roles in inflammation as well as kidney homeostasis. Recent studies suggest that TonEBP expression is also involved in differentiation of several cell types such as myocytes, chondrocytes, and osteocytes. In this study, we investigated the roles of TonEBP during adipocyte differentiation in 3T3-L1 cells. TonEBP mRNA and protein expression was dramatically reduced during adipocyte differentiation. Sustained expression of TonEBP using an adenovirus suppressed the formation of lipid droplets as well as the expression of FABP4, a marker of differentiated adipocytes. TonEBP also inhibited the expression of $PPAR{\gamma}$, a known master regulator of adipocytes. RNAi-mediated knock down of TonEBP promoted adipocyte differentiation. However, overexpression of TonEBP did not affect adipogenesis after the initiation of differentiation. Furthermore, TonEBP expression suppressed mitotic clonal expansion and insulin signaling, which are required early for adipocyte differentiation of 3T3-L1 cells. These results suggest that TonEBP may be an important regulatory factor in the early phase of adipocyte differentiation.

Identification of anti-adipogenic proteins in adult bovine serum suppressing 3T3-L1 preadipocyte differentiation

  • Park, Jeongho;Park, Jihyun;Nahm, Sang-Soep;Choi, Inho;Kim, Jihoe
    • BMB Reports
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    • v.46 no.12
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    • pp.582-587
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    • 2013
  • Adipocyte differentiation is a complex developmental process forming adipocytes from various precursor cells. The murine 3T3-L1 preadipocyte cell line has been most frequently used in the studies of adipocyte differentiation. Differentiation of 3T3-L1 preadipocytes includes a medium containing fetal bovine serum (FBS) with hormonal induction. In this study, we observed that differentiation medium containing adult bovine serum (ABS) instead of FBS did not support differentiation of preadipocytes. Impaired adipocyte differentiation was due to the presence of a serum protein factor in ABS that suppresses differentiation of preadipocytes. Using a proteomic analysis, alpha-2-macroglobulin and paraoxonase/arylesterase 1, which were previously shown to suppress differentiation of preadipocytes, were identified as anti-adipogenic proteins. Although their functional mechanisms have not yet been elucidated, the anti-adipogenic effects of these proteins are discussed.

Inhibitory Effect of Eggplant Extract on Adipocyte Differentiation in 3T3-L1 Cells (가지 물추출물의 3T3-L1 지방전구세포 분화 억제효능)

  • Lee, Mi-Kyeong;Liu, Qing;Hwang, Bang-Yeon;Kim, Sun-Yeou;Lee, Jae-Hak
    • YAKHAK HOEJI
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    • v.55 no.4
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    • pp.309-313
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    • 2011
  • Abnormal growth of adipocyte characterized by increased cell numbers and differentiation is considered as an major pathological characteristic feature in obesity. Thus, inhibition of mitogenesis of preadipocytes and their differentiation to adipocytes would be beneficial for the prevention and inhibition of obesity. In the present study, we attempted to evaluate anti-adipogenic activity of eggplants (the fruits of Solanum melongena L.) employing preadipocytes cell line, 3T3-L1 as an in vitro assay system. Water extract of eggplants significantly inhibited adipocyte differentiation when treated during adipocyte differentiation process, as assessed by measuring fat accumulation using Oil Red O staining. Eggplant extract, however, showed little effects on fully differentiated adipocytes. Eggplant didn't show significant toxicity up to 500 ${\mu}g$/ml to the 3T3-L1 cells. Further studies with interval treatment demonstrated that eggplant exerted inhibitory activity on adipocyte differentiation via acting on early stage of adipogenesis. Conclusively, eggplants are suggested to be beneficial for the prevention of obesity.