基于肺部苦味受体抗哮喘天然活性成分筛选模型的建立与应用

Establishment and application of screening model for anti-asthmatic natural active ingredients based on bitter taste receptors in lung tissue

目的构建以肺部苦味受体TAS2R14为靶点的高通量筛选细胞模型,为寻找高效、低毒的新型抗哮喘天然药物奠定基础。方法利用同源重组技术将TAS2R14受体基因插入p LVX-basic质粒,构建p LVX-Ac GFP1-N1-TAS2R14重组载体,并将其转染入HEK293T细胞,建立特异性高表达TAS2R14受体基因的细胞株。利用该细胞模型对120种苦味中药提取物和单体化合物进行高通量筛选。结果建立的模型Z′因子为0.69和0.66,筛选发现陈皮提取物及其药效物质柠檬苦素,白果提取物及其药效物质芦丁、奎宁对TAS2R14受体具有显著的激动作用。结论建立的TAS2R14受体激动剂高通量筛选模型稳定性好、灵敏度高,筛选得到的3种中药单体具有潜在的TAS2R14受体激动活性。

Objective To construct high-throughput screening cell model targeting TAS2R14 receptor and lay the foundation for the search of effective, novel natural anti-asthma drugs with low toxicity. Methods The pLVX-AcGFPl-N1-TAS2R14 lentivims vector carrying green fluorescent protein （GFP） was constructed. The lentiviral vector was transfected into HEK HEK293T cells, collected high titer lentiviral concentration liquid and infected HEK293T cells, established cell model highly specific expressed TAS2R14 receptor gene. The TAS2R14 cell model was used to screen 120 kinds of Chinese herb extracts and chemical monomers. Results The calculated Z＇ values of the cell model were 0.69 and 0.66, and Citri Reticulatae Pericarpium extract and its efficacy material limonin, Ginkgo Semen extract and its efficacy material mtin and quinine agitated TAS2R14 cell model. Conclusion The constructed TAS2R14 cell model is stable and sensitive for screening anti-asthma drugs, and three kinds of Chinese materia medica monomers have the potential agonist the activity on TAS2R 14 receptor.