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目的:观察灯盏花素对人张氏肝细胞中孕烷X受体(PXR)及细胞色素P450 3A4(CYP3A4)和细胞色素P450 2C19(CYP2C19)mRNA和蛋白表达的影响,初步探讨灯盏花素对CYP3A4和CYP2C19的影响及其作用机制。方法:MTT法检测不同浓度的灯盏花素对张氏肝细胞活力的影响;采用实时荧光定量PCR(RT-qPCR)检测不同浓度灯盏花素对张氏肝细胞中PXR、CYP3A4及CYP2C19 mRNA的表达;采用Western-blot检测不同浓度的灯盏花素对CYP3A4和CYP2C19蛋白的表达影响。结果:MTT结果表明不同浓度的灯盏花素处理张氏肝细胞24 h、48 h后的IC50分别为9 768.8、1 372.8μmol·L~(-1),IC10分别为2 689.2、154.4μmol·L~(-1);RT-qPCR结果表明灯盏花素对CYP3A4和PXR mRNA的影响呈抑制作用,对CYP2C19 mRNA的影响呈不同程度的促进作用;Western-blot结果表明灯盏花素对CYP3A4蛋白的影响呈抑制作用,对CYP2C19蛋白的影响呈促进作用。结论:灯盏花素对CYP3A4的抑制作用可能通过抑制PXR途径;对CYP2C19的促进作用可能不经PXR通路。
OBJECTIVE: To observe the effects of breviscapine on mRNA and protein expression of pregnane X receptor and CYP3A4 and CYP2C19 in human hepatocellular carcinoma cells, and to investigate the effect of breviscapine on Effects of CYP3A4 and CYP2C19 and Their Mechanism. Methods: MTT assay was used to detect the effect of breviscapine on the viability of Zhang’s hepatocytes. Real-time quantitative PCR (RT-qPCR) was used to detect the expression of PXR, CYP3A4 and CYP2C19 mRNA in Zhang hepatocytes The effects of different concentrations of Breviscapine on the expression of CYP3A4 and CYP2C19 protein were detected by Western-blot. Results: The results of MTT assay showed that the IC50 of Zhangzhang hepatocytes treated with different concentrations of breviscapine for 24 h and 48 h were 9 768.8 and 1 372.8 μmol·L -1, respectively, and IC 10 were 2 689.2 and 154.4 μmol·L ~ (-1). The results of RT-qPCR showed that breviscapine inhibited CYP3A4 and PXR mRNA expression and promoted CYP2C19 mRNA expression to a certain extent. The results of Western-blot showed that breviscapine exerted its effects on CYP3A4 protein Was inhibited, the impact of CYP2C19 protein was promoted. Conclusion: Breviscapine can inhibit the CYP3A4 by inhibiting the PXR pathway. The promotion of CYP2C19 may not be through the PXR pathway.