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A possible regulatory link between Twist 1 and PPAR? gene regulation in 3T3-L1 adipocytes.

ABSTRACT: Peroxisome proliferator-activated receptor ? (PPAR?) is a critical gene that regulates the function of adipocytes. Therefore, studies on the molecular regulation mechanism of PPAR? are important to understand the function of adipose tissue. Twist 1 is another important functional gene in adipose tissue, and hundreds of genes are regulated by Twist 1. The aim of this study was to investigate the regulation of Twist 1 and PPAR? expression in 3T3-L1 mature adipocytes.We induced differentiation in 3T3-L1 preadipocytes and examined alterations in Twist 1 and PPAR? expression. We used the PPAR? agonist pioglitazone and the PPAR? antagonist T0070907 to investigate the effect of PPAR? on Twist 1 expression. In addition, we utilized retroviral interference and overexpression of Twist 1 to determine the effects of Twist 1 on PPAR? expression.The expression levels of Twist 1 and PPAR? were induced during differentiation in 3T3-L1 adipocytes. Application of either a PPAR? agonist (pioglitazone) or antagonist (T0070907) influenced Twist 1 expression, with up-regulation of Twist 1 under pioglitazone (1 ?M, 24 h) and down-regulation of Twist 1 under T0070907 (100 ?M, 24 h) exposure. Furthermore, the retroviral interference of Twist 1 decreased the protein and mRNA expression of PPAR?, while Twist 1 overexpression had the opposite effect.There was a possible regulatory link between Twist 1 and PPAR? in 3T3-L1 mature adipocytes. This regulatory link enhanced the regulation of PPAR? and may be a functional mechanism of Twist 1 regulation of adipocyte physiology and pathology.

SUBMITTER: Ren R

PROVIDER: S-EPMC5101646 | biostudies-literature | 2016 Nov

REPOSITORIES: biostudies-literature

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A possible regulatory link between Twist 1 and PPARγ gene regulation in 3T3-L1 adipocytes.

Ren Rui R Chen Zhufeng Z Zhao Xia X Sun Tao T Zhang Yuchao Y Chen Jie J Lu Sumei S Ma Wanshan W

Lipids in health and disease 20161108 1

<h4>Background</h4>Peroxisome proliferator-activated receptor γ (PPARγ) is a critical gene that regulates the function of adipocytes. Therefore, studies on the molecular regulation mechanism of PPARγ are important to understand the function of adipose tissue. Twist 1 is another important functional gene in adipose tissue, and hundreds of genes are regulated by Twist 1. The aim of this study was to investigate the regulation of Twist 1 and PPARγ expression in 3T3-L1 mature adipocytes.<h4>Methods< ...[more]

PMID: 27825360

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Project description:Purpose: Mitochondrial dysfunction in adipose tissue has emerged as key to the development of obesity and diabetes. Salvianolic acid B (SalB) is a water-soluble ingredient derived from Salvia miltiorrhiza that has been shown to possess potential anti-obese and anti-diabetic activities. However, the cellular mechanism of SalB on mitochondrial function with respect to these metabolic disorders has not been elucidated. Therefore, we aim to investigate the effects of SalB on mitochondrial function in 3T3-L1 adipocytes and analyze the underlying molecular mechanism. Methods: The effects of SalB on adipocyte differentiation, glucose uptake, and glycerol release were evaluated in 3T3-L1 adipocytes. Differentiated adipocytes were treated with SalB (50 μM) with or without PPARγ antagonist (GW9662, 20 μM) for 48 h, and mitochondrial oxygen consumption rate (OCR) as well as extracellular acidification rate (ECAR) were assessed using an XF Extracellular Flux Analyzer. The mitochondrial distribution of adipocytes was assessed using Mito Tracker Green (MTG) and observed under a fluorescent microscope. In addition, the mRNA expression levels of peroxisome proliferators-activated receptor γ/α (PPARγ/α), CCAAT/enhancer binding proteinα (C/EBPα), Nuclear respiratory factor 1/2 (NRF1/2), Uncoupling protein 2 (UCP2), and phosphofructokinase 2/fructose-2, 6-bisphosphatase 2 (PFKFB2) were detected by RT-PCR. Finally, changes in the protein levels of peroxisome proliferators-activated receptor γ coactivator-1α (PGC-1α) were determined by western blotting and immunofluorescence analysis. Results: Treatment with SalB increased glucose uptake and mitochondrial respiration, reduced glycerol release and promoted adipocyte differentiation by increasing mRNA expression of adipogenic transcription factors including PPARγ, C/EBPα, and PPARα. Furthermore, SalB enhanced adipocytes mitochondrial content, mitochondrial respiration and glycolysis capacity, which had been attenuated by GW9662 treatment through the increased expression of PGC-1α. Conclusion: Our results provide novel insights into the role of PGC-1α and mitochondria as probable mediators of SalB activity in the regulation of adipocyte differentiation in 3T3-L1 adipocytes.

Dataset Information

A possible regulatory link between Twist 1 and PPAR? gene regulation in 3T3-L1 adipocytes.

Publications

A possible regulatory link between Twist 1 and PPARγ gene regulation in 3T3-L1 adipocytes.

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