Development and evaluation of a cell model targeted on GLP-1 receptor / 中国药理学通报

ABSTRACT

Aim To establish a cell model targeting on GLP-1 R,and evaluate its function by the cAMP assay,for screening the new class of GLP-1 analogues as anti-diabetic candidates. Methods An eukaryotic expression vector pEGFP-GLP-1 R was constructed and transfected into HEK293A cells.After selecting with G41 8,a cell line stably expressing GLP-1 R-GFP was estab-lished.The expression and the cellular distribution of GLP-1 R-GFP fusion protein were investigated by Western blot and fluo-rescence microscopy.Then,the activity of GLP-1 analogue Lira-glutide was evaluated by monitoring the content of cAMP via HTRF using this cell model.Results GLP-1 R-GFP-293A cell line was successfully established.GLP-1 R-GFP fusion proteins were mainly distributed in the cell membrane.The dose-respon-sive relationship experiments revealed that cAMP could be effec-tively stimulated by Liraglutide using this cell model.Conclu-sion This cell model could be used to detect the bioactivity of GLP-1 analogues in vitro,which lays a foundation for the screen-ing of GLP-1 analogues and small GLP-1 R agonists.