Journal
JOURNAL OF ZHEJIANG UNIVERSITY-SCIENCE B
Volume 10, Issue 4, Pages 243-250Publisher
ZHEJIANG UNIV
DOI: 10.1631/jzus.B0820340
Keywords
beta(2)-adrenoceptor; Enhanced green fluorescent protein (EGFP); Chinese medicinal herbs; Screening
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Funding
- National Natural Science Foundation of China [30873103]
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In order to develop a model for screening the agonists of human beta(2)-adrenoceptor from Chinese medicinal herbs extracts, we used a cell-based functional assay based on a common G protein-coupled receptor (GPCR) regulation mechanism and destabilized enhanced green fluorescent protein (d(2)EGFP) reporter gene technique. The positive cell clone was confirmed by real-time polymerase chain reaction (PCR) and imaging analysis. To assess the value of this model, we screened over 2000 high performance liquid chromatography (HPLC)-fractionated samples from the ethanol extracts of Chinese medicinal herbs. Six fractions (isolated from Panax japonicus, Veratrum nigrum, Phellodendron amurense, Fructus Aurantii Immaturus, Chaenomeles speciosa, and Dictamnus dasycarpus) showed significant effects on active reporter gene expression, three of which (isolated from Phellodendron amurense, Fructus Aurantii Immaturus, and Chaenomeles speciosa) were selected for further concentration response analysis and the half maximal effective concentration (EC1/2 max) values were 4.2, 2.7, and 4.8 mu g/ml, respectively. Therefore, this reporter gene assay was suitable for screening beta(2)-adrenoceptor agonists. The results suggest that the six herbal extracts are the possible agonists of beta(2)-adrenoceptor.
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