灯盏花素对张氏肝细胞中CYP3A4和CYP2C19的影响和机制

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、1 372.8 μmol·L^-1，IC₁₀分别为2 689.2、154.4 μmol·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。

Effect of breviscapine on CYP3A4 and CYP2C19 activities and its molecular mechanisms in Chang Liver cells

OBJECTIVE To find out the effects of breviscapine on mRNA and protein expressions of PXR, CYP3A4 and CYP2C19 in Chang Liver cells and preliminarrily study the molecular mechanisms of breviscapine on CYP3A4 and CYP2C19.METHODS The effect of different concentrations of breviscapine on viability of Chang Liver cells was tested by MTT assay. The mRNA and protein expression of PXR, CYP3A4 and CYP2C19 in Chang Liver cells treated with breviscapine were detected by RT-qPCR and Western-blot analysis.RESULTS MTT results showed that the 50% inhibition rates at 24 h and 48 h were 9 768.8 and 1 372.8 μmol·L^-1 respectively when treated with different concentrations of breviscapine in Chang Liver cells for 24 and 48 hours. The 10% inhibition rates at 24 h and 48 h were 2 689.2 and 154.4 μmol·L^-1 when treated with different concentrations of breviscapine in Chang Liver cells for 24 and 48 hours. RT-qPCR results showed that breviscapine inhibited mRNA expression of CYP3A4 and PXR, promoted mRNA expression of CYP2C19. Western-blot results showed breviscapine inhibited protein expression of CYP3A4 and promoted protein expression of CYP2C19.CONCLUSION The inhibitory effect of breviscapine on CYP3A4 may be related with the activation of PXR. The promoting effect of breviscapine on CYP2C19 may not be related with the way of PXR.