以GLP-1受体为靶点的药物筛选细胞模型的建立和应用

建立以GLP-1信号通路为靶点的药物筛选细胞模型，用于筛选GLP-1受体激动剂类的新型抗糖尿病药物。首先构建GLP-1受体信号通路调控的特异应答元件（RIP-CRE）多拷贝序列及报告基因E-GFP的重组载体。将该载体转染胰岛NIT-1细胞株，以检测转染细胞对GLP类似物的反应性和特异性，并通过稳定转染和单克隆培养，获得对GLP-1类似物特异应答的单克隆细胞株。该细胞模型在GLP-1类似物Exendin 4刺激下激活表达报告基因，激活作用可以被GLP-1受体阻断剂Exendin 9-39完全阻断，且激活途径非cAMP-PKA依赖，具有GLP-1受体特异性。构建该细胞模型，可以对肽类或非肽类GLP-1类似物进行高通量筛选。

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.