[A novel cell model targeted on GLP-1 receptor for application to anti-diabetic candidates screening].
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.
Huan
Yi
Y
Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China.
Shen
Zhu-fang
ZF
D000818:Animals; D002460:Cell Line; D051786:Cyclic AMP Response Element Modulator / Q000494:pharmacology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors; D016503:Drug Delivery Systems; D004353:Drug Evaluation, Preclinical; D000077270:Exenatide; D017930:Genes, Reporter; D000067757:Glucagon-Like Peptide-1 Receptor; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007004:Hypoglycemic Agents / Q000819:agonists / Q000037:antagonists & inhibitors / Q000378:metabolism; D007515:Islets of Langerhans / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D007546:Isoquinolines / Q000494:pharmacology; D010446:Peptide Fragments / Q000494:pharmacology; D010455:Peptides / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D010957:Plasmids; D051381:Rats; D018027:Receptors, Glucagon / Q000819:agonists / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D013449:Sulfonamides / Q000494:pharmacology; D014162:Transfection; D014688:Venoms / Q000494:pharmacology*
C000593835:Glp1r protein, rat; D000067757:Glucagon-Like Peptide-1 Receptor; D007004:Hypoglycemic Agents; D007546:Isoquinolines; D010446:Peptide Fragments; D010455:Peptides; D018027:Receptors, Glucagon; D011994:Recombinant Proteins; D013449:Sulfonamides; D014688:Venoms; C485184:enhanced green fluorescent protein; D051786:Cyclic AMP Response Element Modulator; D049452:Green Fluorescent Proteins; C083773:exendin (9-39); D000077270:Exenatide; D017868:Cyclic AMP-Dependent Protein Kinases; C063509:N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide