Establishment of a cell-based high-throughput screening model for PPARdelta agonists.
- Author:
Guo-Ping LU
1
;
Zhi-Hui ZHENG
;
Bao-Hua ZHAO
;
Si-Hai XIANG
;
Yue-Sheng DONG
;
Fu-Qiang WANG
;
Hua ZHANG
Author Information
1. College of Life Sciences, Hebei Normal University, Shijiazhuang 050016, China.
- Publication Type:Journal Article
- MeSH:
3T3 Cells;
Animals;
Bezafibrate;
pharmacology;
Cell Line;
Dose-Response Relationship, Drug;
Drug Evaluation, Preclinical;
methods;
Genetic Vectors;
chemistry;
genetics;
HeLa Cells;
Humans;
Linoleic Acid;
pharmacology;
Lipids;
chemistry;
Luciferases;
genetics;
metabolism;
Mice;
PPAR delta;
agonists;
genetics;
Recombinant Fusion Proteins;
genetics;
metabolism;
Transfection;
methods
- From:
Chinese Journal of Biotechnology
2007;23(2):343-346
- CountryChina
- Language:Chinese
-
Abstract:
To establish a new high-throughput screening model for the agonist of PPARdelta, PPARdelta gene was obtained by reverse transcriptase-polymerase chain reaction (RT-PCR), and subcloned to pGEM-T Vector for sequencing, then the PPARdelta fragment was excised by restriction enzymes, and inserted into pTARGET Vector to construct expression vector pTARGET-ppARdelta. Insert three copies of PPRE into pGl3-promoter vector to construct expression vector pGl3-PPRE x 3-luc. The vector pTARGET-ppARdelta was transiently cotransfected with pGl3-PPRE x 3-luc into different cell lines to assay the expression levels of luciferase. The PPARdelta agonist screening model was established and optimized. Bezafibrate and linoleic acid can induce the expression of luciferase significantly and in a dose-dependent manner. This method can be used for high throughput screening for the agonist of PPARdelta, which might become lead compounds for new anti-atheroscleriosis or anti-adiposity drugs.
