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 GLUT 1-mediated glucose uptake. This novel cell model can be conveniently used in high-throughput drug screening of FGF-21 or FGF-21 analogues.%建立以成纤维细胞生长因子-21(fibroblast growthfactor-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βklotho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βklotho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βklotho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.
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