中国生物工程杂志

ABSTRACT: AIM A cell based reporter gene assay had been established and optimized for high throughput screening to find agonists for human histamine 3 receptor (H3R) from Chinese herb components. METHODS The H3 receptor plasmid (H3R/pCDNA3.1-hygro) and reporter gene plasmid (3XCRE-LUC) were cotransfected HEK293 and a stable cell line was selected with antibiotics for H3 receptor agonist screening. Upon agonist binding, H3 receptor was activated and lead to a decrease of the high expression of luciferase gene induced by forskolin. RESULT To identify and optimize the assay condition, the effects of some factors were examined, such as compound solvent selection, final concentration of forskolin, agonist and forskolin incubation time, final concentration of DMSO. A steady cell line and a reliable method were established for H3R agonist screening, the Z’factor value was above 0.5, the signal did not change using HEK/H3R/CRE cell line passage 5 to passage 20. By using this assay system, two herb components were identified and regarded to contain agonists for H3 receptor. CONCLUSION This drug screening cell model has been successfully used for histamine 3 receptor target high throughput drug screening.