1. College of Medical Technology, Beihua University, jilin 132013, China; 2. Jilin University of Medicine, Jilin 132013, China; 3. Medical College of Yanbian University, Yanji 133002, China
Abstract：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-H148Q/I152L 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-H148Q/I152L 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.
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2022, Vol. 57 
