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目的研究6种中药单体野黄芩苷、丹皮酚、芍药苷、甘草苷、黄芪甲苷以及黄芩素是否可以通过诱导孕烷X受体(PXR)介导细胞色素酶P4503A4(CYP3A4)转录表达。方法采用脂质体瞬时转染的方法,将人PXR表达质粒、CYP3A4报告质粒以及内参质粒转入LS174T细胞中,建立报告基因体系,与不同浓度及不同给药时间的中药单体共同孵育,检测细胞中荧光素酶的活性。结果黄芩素(20μmol/L、40μmol/L和80μmol/L)分别诱导PXR介导CYP3A4表达(1.49±0.23)倍(P<0.05)、(2.45±0.56)倍(P<0.01)和(5.27±0.40)倍(P<0.01),野黄芩苷、丹皮酚、芍药苷、甘草苷和黄芪甲苷在实验的浓度范围内不能明显诱导LS174T细胞中PXR介导CYP3A4表达。在不同给药时间实验中,20μmol/L、40μmol/L和80μmol/L的黄芩素在12～48h能诱导PXR介导CYP3A4表达,诱导能力随时间的延长而呈增强的趋势,各浓度的最大诱导倍数均在48h出现。结论黄芩素在LS174T细胞系中可以通过诱导PXR介导CYP3A4转录表达,而野黄芩苷、丹皮酚、芍药苷、甘草苷或黄芪甲苷无此作用。
Objective To investigate whether six Chinese herbal medicines, baicalin, paeonol, paeoniflorin, liquiritin, astragaloside and baicalein could mediate the transcription of cytochrome P4503A4 (CYP3A4) through the induction of pregnane X receptor expression. Methods The human PXR expression plasmid, CYP3A4 reporter plasmid and the internal control plasmid were transfected into LS174T cells by lipofection method. The reporter gene system was established and incubated with Chinese medicine monomer with different concentrations and different administration time. Luciferase activity in cells. Results Baicalein (20μmol / L, 40μmol / L and 80μmol / L) induced PXR-mediated CYP3A4 expression by 1.49 ± 0.23 fold (2.45 ± 0.56) fold 0.40) fold (P <0.01). Scutellarin, paeonol, paeoniflorin, glycyrrhizin and astragaloside did not significantly induce PXR-mediated CYP3A4 expression in LS174T cells in the experimental concentration range. At different dosing time, baicalein at 20μmol / L, 40μmol / L and 80μmol / L induced PXR-mediated CYP3A4 expression at 12-48h, and the inducing ability tended to increase with time Induction multiple appear in 48h. Conclusion Baicalein can induce PXR-mediated transcription of CYP3A4 in LS174T cell line without baicalin, paeonol, paeoniflorin, glycyrrhizin or astragaloside.