Impact of rosiglitazone on the expression of beta3-AR in the stable cell lines expressed beta3-AR gene.
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.
Zhao
Xin
X
Pharmacogenetics Research Institute, Institute of Clinical Pharmacology, Central South University Xiang-Ya School of Medicine, Changsha, Hunan, P.R. China.
Hu
Min-Yu
MY
Huang
Qiong
Q
Tang
Zhong-Yang
ZY
Dai
Xing-Ping
XP
Yin
Ji-Ye
JY
Zhang
Feng
F
Zhou
Hong-Hao
HH
Liu
Zhao-Qian
ZQ
D001483:Base Sequence; D015153:Blotting, Western; D045744:Cell Line, Tumor; D017931:DNA Primers; D006801:Humans; D007004:Hypoglycemic Agents / Q000494:pharmacology*; D047495:PPAR gamma / Q000235:genetics; D010957:Plasmids; D022702:Receptors, Adrenergic, beta-3 / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000077154:Rosiglitazone; D045162:Thiazolidinediones / Q000494:pharmacology*
D017931:DNA Primers; D007004:Hypoglycemic Agents; D047495:PPAR gamma; D022702:Receptors, Adrenergic, beta-3; D045162:Thiazolidinediones; D000077154:Rosiglitazone