Study on Cell Model for High Throughput Screening the Agonist of Glucagon-Like Petide-1 Receptor

In order to establish high throughout screening method for identifying agonists for glucagon-like petide-1 receptor(GLP-1R),a cell line was set up after the GLP-1 receptor plasmid(GLP-1R/pCDNA3.1) and reporter gene plasmid(3×CRE/3×MRE/SRE-LUC/ pGL3)were co-transfectted CHO at the ratio of 1∶5.To identify and optimize the screening model,the effects of some factors were examined by using GLP-1,such as cell number per well,incubation time,dosage of Bright-GloTM and concentration of DMSO.Based on a steady cell line,a reliable screening model was established for GLP-1R agonist screening.The Z′-factor value for the screening model was estimated 0.75 under the conditions that 4×104 cells in each well were incubated for 8h using 4 times diluted Bright-GloTM and less than 1% DMSO.The extracts of tradition Chinese medicine were assayed using the screening model,and five samples of them showed activity.Experiments showed that this method could be applied to high throughout screen agonist for GLP-1 receptor.