Evaluation of Reference Genes for Studies of Gene Expression in Human Adipose Tissue

Gabrielsson, Britt G.; Olofsson, Louise E.; Sjögren, Anders; Jernås, Margareta; Elander, Anna; Lönn, Malin; Rudemo, Mats; Carlsson, Lena

doi:10.1038/oby.2005.72

Cited by 115 publications

(126 citation statements)

References 13 publications

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“…Amplification efficiencies were validated and normalized to ribosomal 18S and quantity of target gene was calculated according to a standard curve. 18S is an established and accepted reference gene (14). Our own experience showed that it is comparable with other reference genes including LRP10, CLN3, and HPCAL1 (data not shown).…”

Section: Zfp36 Measurementsmentioning

confidence: 69%

Visceral adipose tissue zinc finger protein 36 mRNA levels are correlated with insulin, insulin resistance index, and adiponectinemia in women

Bouchard¹,

Vohl²,

Deshaies³

et al. 2007

eur j endocrinol

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Introduction: Adipose tissue is now recognized as an endocrine organ and secretes numerous molecules and proteins potentially involved in the physiopathology of the metabolic syndrome. Recently, we have determined the transcriptome of omental adipose tissue, leading to the identification of a new candidate gene for obesity-related metabolic complications, zinc finger protein 36 (ZFP36), which is known to down-regulate tumor necrosis factor-a (TNF-a) expression. Objective: The objective of this study was to further examine the relationship between ZFP36 gene expression levels, obesity-related phenotypes, and adipokines. Methods: Abdominal subcutaneous and omental adipose tissue samples were obtained from 46 women undergoing elective gynecological surgery. Adipose tissue ZFP36 mRNA abundance was assessed by quantitative real-time PCR. Body fat accumulation and distribution were measured by dual X-ray absorptiometry and computed tomography. Fasting blood levels of glucose, insulin, and lipids, and circulating TNF-a, interleukin-6 (IL-6), resistin, and adiponectin were also measured. Results: No correlation was observed between s.c. ZFP36 mRNA levels and any of the phenotypes tested. However, although omental ZFP36 mRNA levels were not correlated with measures of body fatness and lipid profile, they were negatively correlated with fasting insulin levels (RZK0.31; PZ0.05), the insulin resistance index (HOMA-IR; RZK0.31; PZ0.05), and 2-h post-glucose insulinemia (RZK0.32; PZ0.05). Omental ZFP36 mRNA abundance was also positively correlated with adiponectinemia (RZ 0.35; PZ0.03) but not with circulating TNF-a, IL-6, and resistin concentrations. Conclusion: These results suggest that ZFP36 gene expression in omental adipose tissue, but not in abdominal s.c. fat, may offer partial protection against the development of insulin resistance and diabetes.

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Section: Zfp36 Measurementsmentioning

confidence: 69%

Visceral adipose tissue zinc finger protein 36 mRNA levels are correlated with insulin, insulin resistance index, and adiponectinemia in women

Bouchard¹,

Vohl²,

Deshaies³

et al. 2007

eur j endocrinol

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“…C t values were normalized to the reference gene, low-density lipoprotein receptor-related protein 10 (LRP10), which was amplified in parallel reactions. LRP10 was preferred as a reference gene because it displayed the least variation in expression among a number of reference genes in human adipose tissue (25). RNA interference of IRS-1.…”

Section: Methodsmentioning

confidence: 99%

Vascular Peptide Endothelin-1 Links Fat Accumulation With Alterations of Visceral Adipocyte Lipolysis

et al. 2008

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OBJECTIVE-Visceral obesity increases risk of insulin resistance and type 2 diabetes. This may partly be due to a regionspecific resistance to insulin's antilipolytic effect in visceral adipocytes. We investigated whether adipose tissue releases the vascular peptide endothelin-1 (ET-1) and whether ET-1 could account for regional differences in lipolysis.RESEARCH DESIGN AND METHODS-One group consisted of eleven obese and eleven nonobese subjects in whom ET-1 levels were compared between abdominal subcutaneous and arterialized blood samples. A second group included subjects undergoing anti-obesity surgery. Abdominal subcutaneous and visceral adipose tissues were obtained to study the effect of ET-1 on differentiated adipocytes regarding lipolysis and gene and protein expression.RESULTS-Adipose tissue had a marked net release of ET-1 in vivo, which was 2.5-fold increased in obesity. In adipocytes treated with ET-1, the antilipolytic effect of insulin was attenuated in visceral but not in subcutaneous adipocytes, which could not be explained by effects of ET-1 on adipocyte differentiation. ET-1 decreased the expression of insulin receptor, insulin receptor substrate-1 and phosphodiesterase-3B and increased the expression of endothelin receptor-B (ET B R) in visceral but not in subcutaneous adipocytes. These effects were mediated via ET B R with signals through protein kinase C and calmodulin pathways. The effect of ET-1 could be mimicked by knockdown of IRS-1. A bdominal obesity contributes to the pathogenesis of insulin resistance and, thereby, type2 diabetes (1,2). Release of free fatty acids (FFAs) from fat cells during lipolysis may be involved in the negative consequences of excess adipose tissue (3-5). This process is inhibited by insulin, which activates a signaling pathway including insulin substrate (IRS)-1, phosphatidylinositol 3-kinase (PI3K), AKT, and, ultimately, the enzyme phosphodiesterase-3B (PDE3B), which breaks down cyclic AMP. There are important regional variations in the antilipolytic effect of insulin. Subcutaneous adipocytes are much more sensitive than visceral adipocytes because of a higher receptoraffinity and higher expression of IRS-1 (6 -8). Site variations in adipocyte lipolysis elevate release of FFAs from the visceral compared with the subcutaneous adipose tissue during hyperinsulinemia (e.g., postprandially). Only visceral fat is linked to the liver, and a high FFA mobilization to the liver results in hepatic insulin resistance, dyslipidemia, hyperglycemia, and hyperinsulinemia, all of which are features of type 2 diabetes (3-5). CONCLUSIONS-ET-The factors that determine the relative insulin sensitivity of various adipose tissue depots are not well understood, but local environment may contribute. Adipocytes are surrounded by stromal vascular cells, including endothelial cells, which secrete endothelin-1 (ET-1), a potent vasoconstrictor. Plasma levels of ET-1 are increased in obesity and type 2 diabetes (9 -13), although the major source of circulating ET-1 in these conditions is not ...

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“…A standard curve was plotted for each primer-probe set with a serial dilution of pooled adipose tissue cDNA. Human LRP10 [29] was used as a reference to normalize the expression levels between the samples. All the standards and samples were analyzed in triplicate.…”

Section: Rt-pcr Analysismentioning

confidence: 99%

Regulation of human aldoketoreductase 1C3 (AKR1C3) gene expression in the adipose tissue

Svensson

Gabrielsson

Jernås

et al. 2008

Cellular and Molecular Biology Letters

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Aldoketoreductase 1C3 (AKR1C3) is a functional prostaglandin F synthase and a negative modulator of the availability of ligands for the nuclear receptor peroxisome proliferator-activated receptor-gamma (PPARγ). AKR1C3 expression is known to be associated with adiposity, one of the components of the metabolic syndrome. The aim of this study was to characterize the expression of AKR1C3 in the adipose tissue and adipocytes and to investigate its potential role in the metabolic syndrome. Using microarray analysis and realtime PCR, we studied the expression of AKR1C3 in adipose tissue samples from obese subjects with or without metabolic complications, during very low calorie diet-induced weight loss, and its expression in isolated human adipocytes of different sizes. The adipose tissue AKR1C3 expression levels were marginally lower in obese subjects with the metabolic syndrome compared with the levels in healthy obese subjects when analyzed using microarray (p = 0.078) and realtime PCR (p < 0.05), suggesting a secondary or compensatory effect. The adipose tissue mRNA levels of AKR1C3 were reduced during and after dietinduced weight-loss compared to the levels before the start of the diet (p < 0.001 at all time-points). The gene expression of AKR1C3 correlated with both adipose tissue mRNA levels and serum levels of leptin before the start of the diet (p < 0.05 and p < 0.01, respectively). Furthermore, large adipocytes displayed a higher expression of AKR1C3 than small adipocytes (1.5-fold, p < 0.01). In conclusion, adipose tissue AKR1C3 expression may be affected by metabolic disease, and its levels are significantly reduced in response to dietinduced weight loss and correlate with leptin levels.

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Evaluation of Reference Genes for Studies of Gene Expression in Human Adipose Tissue

Cited by 115 publications

References 13 publications

Visceral adipose tissue zinc finger protein 36 mRNA levels are correlated with insulin, insulin resistance index, and adiponectinemia in women

Visceral adipose tissue zinc finger protein 36 mRNA levels are correlated with insulin, insulin resistance index, and adiponectinemia in women

Vascular Peptide Endothelin-1 Links Fat Accumulation With Alterations of Visceral Adipocyte Lipolysis

Regulation of human aldoketoreductase 1C3 (AKR1C3) gene expression in the adipose tissue

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