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目的:基于二型大麻受体(CB2)的信号传递通路,建立体外高通量筛选CB2受体激动剂的药物模型。方法:将目的基因质粒pIRES2-EGFP-CB2、报告基因质粒pGL4.29[luc2P/CRE/Hygro]、内参质粒PRL-TK共转染CHO细胞,通过检测双荧光素酶报告基因的表达水平变化来筛选人CB2受体的激动剂。并对加入激动剂的浓度和作用时间进行优化及考察模型的稳定性。结果:给予10μmol/L JWH-015 6 h后能取得最大相对诱导率。成功建立CB2受体激动剂的高通量筛选模型,筛选模型Z’因子为0.81,表现为良好的稳定性。结论:成功地建立了CB2受体激动剂的筛选模型,为从中药中高通量筛选有效物质奠定了良好的基础。
OBJECTIVE: To establish a high-throughput screening model of CB2 receptor agonist in vitro based on the signal transduction pathway of type II cannabinoid receptor (CB2). METHODS: CHO cells were co-transfected with the target gene plasmid pIRES2-EGFP-CB2, the reporter plasmid pGL4.29 [luc2P / CRE / Hygro] and the reference plasmid PRL-TK to detect the expression level of the dual luciferase reporter gene Screen human CB2 receptor agonists. The concentration of agonist and the duration of action were optimized and the stability of the model was investigated. Results: The maximum relative induction rate of JWH-015 after 10μmol / L administration for 6 h was obtained. A high-throughput screening model of CB2 receptor agonist was successfully established. The Z ’factor of the screening model was 0.81, showing good stability. Conclusion: The screening model of CB2 receptor agonist has been successfully established, which lays a good foundation for the screening of effective substances by high-throughput in Chinese medicine.