The aim of this project is to establish a GLP-1 signaling pathway targeted cell model, for screening the new class of GLP-1 receptor agonists as anti-diabetic candidates. Firstly construct a recombined plasmid with multi-copied specific response element (RIP-CRE) regulated by GLP-1 signaling pathway and E-GFP reporter gene. Transient transfect this recombined plasmid into islet cell NIT-1, then detect the responsibility of transfected cell to GLP-1 analogue, Exendin 4. For secondly, use stable transfection and monocloning cell culture to obtain a GLP-1 signaling-specific cell line. It indicates that this cell model can response to Exendin 4, which response can be completely inhibited by GLP-1 receptor antagonist, Exendin 9-39, further showing GLP-1 receptor specific activity with a cAMP-PKA-independently mechanism. Establishment of this novel cell model can be used in high-throughput drug screening of peptides or small molecular GLP-1 analogues.
Animals ; Cell Line ; Cyclic AMP Response Element Modulator ; pharmacology ; Cyclic AMP-Dependent Protein Kinases ; antagonists & inhibitors ; Drug Delivery Systems ; Drug Evaluation, Preclinical ; Genes, Reporter ; Glucagon-Like Peptide-1 Receptor ; Green Fluorescent Proteins ; metabolism ; Hypoglycemic Agents ; agonists ; antagonists & inhibitors ; metabolism ; Islets of Langerhans ; cytology ; drug effects ; metabolism ; Isoquinolines ; pharmacology ; Peptide Fragments ; pharmacology ; Peptides ; antagonists & inhibitors ; pharmacology ; Plasmids ; Rats ; Receptors, Glucagon ; agonists ; antagonists & inhibitors ; genetics ; metabolism ; Recombinant Proteins ; genetics ; metabolism ; Signal Transduction ; Sulfonamides ; pharmacology ; Transfection ; Venoms ; pharmacology