Glucagon-like peptide-1 receptor (GLP-1R), glucose-dependent insulinotropic polypeptide receptor (GIPR), and G protein-coupled receptor 40 (GPR40) are members of G protein-coupled receptors (GPCR) family. They are abundantly expressed in islet beta cells, and mediate effects of incretins and fatty acids in beta cells. Glucose and 5-AMP-activated protein kinase (AMPK) are known to be involved in the regulation of beta cell function. Metformin and the potential therapeutic drug for type 2 diabetes, 5-amino-4-imidazolecarboxamide riboside (AICAR), are both known activators of AMPK. Here we studied the effects of glucose, metformin, and AICAR on the expression of GPCR in INS-1 beta cell. INS-1 beta cells were supplemented with different concentrations of glucose, metformin, or AICAR. The expressions of GLP-1R, GIPR, GPR40, and a nuclear transcription factor - peroxisome-proliferator activated receptor alpha (PPARalpha) - were analyzed by real-time RT-PCR and immunoblotting. The time-course of the mRNA degradation of these receptors was also monitored by applying actinomycin D to cells. We demonstrated that the expressions of GLP-1R, GIPR, and PPARalpha were downregulated when INS-1beta cells were treated with glucose, while their expressions were upregulated when treated with metformin or AICAR. Glucose, metformin, or AICAR treatment had no obvious effect on the expression of GPR40. These results indicate that glucose, metformin, and AICAR regulated the expressions of incretin receptors and PPARalpha, but not GPR40 in beta cells. Whether AMPK is a key regulator of these factors mediated receptor regulation remains to be investigated further.