title: Development of a cell-based high-throughput peroxisome proliferator-activated receptors (PPARs) screening model and its application for evaluation of the extracts from Rhizoma Coptis.
abstract: To date, peroxisome proliferator-activated receptors (PPARs) are becoming the new therapeutic targets for the treatment of metabolic diseases, such as Type 2 diabetes, obesity, and cardiovascular disease. In this study, a cell-based high-throughput PPARs (PPARα/β/γ) model was developed for the screening of PPARs agonists. The screening conditions were evaluated through analyzing the expression value of luciferase. Finally, 24 h of drug acting time, 5 times of the dilution factor of luciferase zymolyte, and about 2 × 10(4) cells/ well on HeLa cells in 96-well plates were used, respectively. Furthermore, the quality of high-throughput screening (HTS) in stability and reliability was evaluated by the Z'-factor. Additionally, different extracts of Rhizoma Coptis and berberine were tested by the developed method. The results suggested that both the EtOAc extract and berberine were able to activate PPARα/β/γ, and Rhizoma Coptis contains potential natural agonists of PPARs besides berberine. In conclusion, the developed HTS assay is a simple, rapid, stable, and specific method for the screening of PPARs natural agonists.
authors: College of Bioengineering, Chongqing University, Chongqing 400030, China. chem_lab_cqu@yahoo.com.cn
authors: Zhi-Ning
authors: ZN
authors: Xia
authors: Ye-Xin
authors: YX
authors: Lin
authors: Li-Xia
authors: LX
authors: Guo
authors: Feng-Qing
authors: FQ
authors: Yang
authors: Pan
authors: P
authors: Xu
authors: Yong-Lan
authors: YL
authors: Zhang
authors: Jian-Hui
authors: JH
authors: Liu
mesh_terms: D001599:Berberine / Q000494:pharmacology*; D039323:Coptis / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D008954:Models, Biological; D047493:PPAR alpha / Q000378:metabolism; D047495:PPAR gamma / Q000378:metabolism; D048068:PPAR-beta / Q000378:metabolism; D047492:Peroxisome Proliferator-Activated Receptors / Q000819:agonists*
chemical_list: D047493:PPAR alpha; D047495:PPAR gamma; D048068:PPAR-beta; D047492:Peroxisome Proliferator-Activated Receptors; D001599:Berberine; D008156:Luciferases