Impact of rosiglitazone on the expression of beta(3)-AR in the stable cell lines expressed beta(3)-AR gene.

摘要

Background: The aim of the present study was to investigate the effect of rosiglitazone, a peroxisome proliferator-activated receptor gamma2 (PPAR-gamma2) agonist, on the expression of beta(3)-adrenergic receptor (beta(3)-AR) at transcriptional and translational level. Methods: We cloned the cDNA sequences of human PPAR-gamma2 (hPPAR-gamma2) gene and human wild type and mutant beta(3)-adrenergic receptor (hbeta(3)-AR) genes and established their eukaryotic expression vectors. The pcDNA3.1/hbeta(3)-AR (mutant and wild type) was transfected into SH-SY5Y cells using electroporation method. The expression level of beta(3)-AR protein was determined by Western blot analysis. Results: Our results showed that the reverse transcription-PCR products were consistent with theoretical fragment sizes of human PPAR-gamma2 (1544 bp) and human beta(3)-AR genes (1578 bp). The sequence analysis of PPAR-gamma2 and beta(3)-AR genes showed that the fragment sizes were the same as that of human PPAR-gamma2 and human beta(3)-AR genes in Genebank. The pcDNA3.1/hbeta(3)-AR (mutant and wild type) was successfully cloned to SH-SY5Y cells. We found that the expression of beta(3)-AR protein was significantly inhibited by rosiglitazone in a concentration-dependent manner in SH-SY5Y cell lines stably expressed beta(3)-AR genes. Conclusions: The results suggest that rosiglitazone has a concentration-dependent inhibitory effect on the expression of beta(3)-AR protein, and this inhibitory effect may be due to activation of PPAR-gamma2 receptor. Clin Chem Lab Med 2007;45:1511-6.