筛选糖尿病候选药物FGF-21受体激动剂的新型细胞模型的建立
Establishment of a novel cell model targeted on FGF-21 receptor for screening anti-diabetic drug candidates
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摘要:
建立以成纤维细胞生长因子-21 (fibroblast growth factor-21, FGF-21) 信号通路为靶点的药物筛选细胞模型, 用于筛选FGF-21受体激动剂类的新型治疗糖尿病药物。FGF-21的生理功能主要是通过细胞表面的FGFR及辅助受体β klotho传递信号, 从而激活细胞内相关调控的一系列信号通路以及基因转录来实现的。本实验将β klotho基因构建到逆转录病毒表达载体pBMN-IRES-EGFP。将该载体导入包装细胞, 收集病毒上清液并感染3T3-L1细胞, 筛选到稳定表达β klotho细胞系。该细胞模型在FGF-21作用下可以促进细胞葡萄糖转运蛋白-1表达及转运水平升高, 从而提高细胞糖吸收能力。构建该细胞系可以方便对FGF-21及类似药物进行高通量筛选, 为糖尿病药物的研究提供了一种新方法。
Abstract:The aim of this project is to establish a fibroblast growth factor-21 (FGF-21) signaling pathway targeted cell model, for screening a class of FGF-21 receptor agonists as anti-diabetic candidates. FGF-21 requires β klotho transmembrane proteins as co-receptor for the activation of tyrosine kinase FGF receptor (FGFR) signaling, thereby activating a series of intracellular signaling pathways and regulating gene transcription for glucose metabolism. Firstly a recombinant plasmid expressing co-receptor β klotho and EGFP reporter genes was constructed. After introducing the recombinant plasmid into package cells, the cell culture supernatant was used to infect 3T3-L1 cells, which were then screened for stably expressing β klotho gene. Administration of FGF-21 increased the expression of GLUT1 and stimulated GLUT1-mediated glucose uptake. This novel cell model can be conveniently used in high-throughput drug screening of FGF-21 or FGF-21 analogues.
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