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Diabetologia

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01-05-2007 | Article

Characterisation of insulin-resistant phenotype of cultured rat primary adipose cells

Authors: C. C. Xiang, Y. J. Wu, L. Ma, L. Ding, I. Lisinski, M. J. Brownstein, S. W. Cushman, X. Chen

Published in: Diabetologia | Issue 5/2007

Abstract

Aims/hypothesis

We characterised insulin resistance, metabolic defects and endocrine dysfunction in cultured adipose cells and examined the autocrine or paracrine roles of cytokines/adipokines in the progression of insulin resistance.

Materials and methods

Rat primary adipose cells were prepared and cultured for 24 and 48 h. Insulin resistance and gene expression were examined by glucose uptake assay, cDNA microarray and real-time RT-PCR.

Results

After 24 h in culture, the fold increase of insulin-stimulated glucose uptake in adipose cells was markedly reduced; after 48 h the response of the cells to insulin decreased. cDNA microarray analysis showed that the expression of 514 genes was altered in adipose cells after 24 h in culture. The dysregulated genes included those involved in the citric acid cycle and in fatty acid and pyruvate metabolism. Specifically, the following genes were all downregulated: genes encoding lipolytic and lipogenic enzymes; uncoupling protein 1 and 2 genes; peroxisome proliferator-activated receptor gamma, coactivator 1 alpha gene. This indicates that lipolytic and lipogenic activity, as well as mitochondria capacity decline in adipose cells cultured for 24 h. The mRNAs encoding 40 adipokines were also dysregulated in cultured cells. Strikingly, the dysregulated adipokines in cultured cells and in freshly isolated adipose cells from insulin-resistant Zucker fa/fa rats displayed a similar pattern with regard to protein functions. Also striking was the fact that progression of insulin resistance was promoted by the adipokines secreted from insulin-resistant adipose tissue or cells.

Conclusions/interpretation

Our data demonstrate that the impairment of metabolism and endocrine dysfunction in cultured adipose cells mimics the insulin resistance occurring in vivo. Cytokines and adipokines appear to play a critical role in the progression of insulin resistance in adipose cells.

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Title: Characterisation of insulin-resistant phenotype of cultured rat primary adipose cells
Authors: C. C. Xiang
Y. J. Wu
L. Ma
L. Ding
I. Lisinski
M. J. Brownstein
S. W. Cushman
X. Chen
Publication date: 01-05-2007
Publisher: Springer-Verlag
Published in: Diabetologia / Issue 5/2007
Print ISSN: 0012-186X
Electronic ISSN: 1432-0428
DOI: https://doi.org/10.1007/s00125-007-0626-z

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Abstract

Aims/hypothesis

Materials and methods

Results

Conclusions/interpretation

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