Impact of rosiglitazone on the expression of beta3-AR in the stable cell lines expressed beta3-AR gene.

ABSTRACT

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 beta3-adrenergic receptor (beta3-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 beta3-adrenergic receptor (hbeta3-AR) genes and established their eukaryotic expression vectors. The pcDNA3.1/hbeta3-AR (mutant and wild type) was transfected into SH-SY5Y cells using electroporation method. The expression level of beta3-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 beta3-AR genes (1578 bp). The sequence analysis of PPAR-gamma2 and beta3-AR genes showed that the fragment sizes were the same as that of human PPAR-gamma2 and human beta3-AR genes in Genbank. The pcDNA3.1/hbeta3-AR (mutant and wild type) was successfully cloned to SH-SY5Y cells. We found that the expression of beta3-AR protein was significantly inhibited by rosiglitazone in a concentration-dependent manner in SH-SY5Y cell lines stably expressed beta3-AR genes. CONCLUSIONS: The results suggest that rosiglitazone has a concentration-dependent inhibitory effect on the expression of beta3-AR protein, and this inhibitory effect may be due to activation of PPAR-gamma2 receptor.