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目的:利用分子生物技术构建色氨酸羟化酶(TPH2)启动子调控红色荧光蛋白(red fluorescent prote in,RFP)转基因细胞,以筛选吴茱萸汤及其各组成药物对TPH2启动子的调控活性,探讨其治疗偏头痛可能的分子靶点。方法:构建TPH2启动子调控红色荧光蛋白表达的转基因细胞系,观察统计不同给药组中细胞荧光强度的变化及发光细胞数量的变化。结果:通过荧光细胞计数发现吴茱萸汤、吴茱萸、生姜、大枣水提物不同透析部分在不同浓度上对TPH2启动子转录活性具有不同的调控作用。结论:吴茱萸汤治疗偏头痛的可能分子靶点与激活TPH2表达有关。TPH2启动子调控红色荧光蛋白表达细胞模型可用于对以此为靶点的药物筛选和活性评价。
OBJECTIVE: To construct tryptophan hydroxylase (TPH2) promoter to regulate red fluorescent protein (RFP) transgenic cells by molecular biotechnology, and to screen the regulatory activity of Wuzhi decoction and its constituent drugs on TPH2 promoter. Explore its possible molecular targets for the treatment of migraine. METHODS: The TPH2 promoter was used to construct a transgenic cell line that regulates the expression of red fluorescent protein. The changes of fluorescence intensity and the number of light-emitting cells in different groups were observed and counted. RESULTS: By fluorescence cell counting, it was found that the different dialysis fractions of Wuqitang, Wuqin, Ginger, and Jujube aqueous extracts had different regulatory effects on the transcriptional activity of TPH2 promoter at different concentrations. Conclusion: The possible molecular target of migraine for Wujing Decoction is related to the activation of TPH2 expression. The TPH2 promoter-regulated red fluorescent protein expression cell model can be used to target drug screening and activity evaluation.