基于CFTR的H2受体调节剂高通量筛选细胞模型的构建

目的 建立基于囊性纤维化跨膜传导调节因子(cystic fibrosis transmembrane conductance regulator,CFTR)高通量筛选组胺H2受体(histocompatibility-2)调节剂的模型.方法 应用RT-PCR、Western blot检测H2受体在Fischer大鼠甲状...

Construction of High Throughput Screening Cell Model for H2 Receptor Modulators Based on CFTR

OBJECTIVE To establish a high-throughput screening model of histamine H2 receptor regulators based on cystic fibrosis transmembrane conduction regulator(CFTR). METHODS RT-PCR and Western blot were used to detect the expression of H2 receptor in thyroid follicular epithelial(FRT) cells of Fischer rats. FRT cells expressing CFTR and YFP-H148 Q/I152 L were transfected with liposomes. Fluorescence quenching kinetics test was used to detect the function of the cell model to determine whether the model could screen regulators of H2 receptor. RESULTS RT-PCR and Western blot showed that FRT endogenous expressed H2 receptor. Fluorescence microscopy confirmed that CFTR and YFP-H148 Q/I152 L cell models were successfully constructed. The fluorescence quenching kinetics experiments showed that the model had the characteristics of cAMP activated chloride channels, and could be used to screen H2 receptor modulators. The slope of fluorescence was dose-dependent with the concentration of H2 receptor modulators. CONCLUSION The high-throughput screening cell model of H2 receptor modulators based on CFTR is successfully constructed.