黄连素通过PXR信号通路调控CYP3A4和P-gp的机制研究

Mechanism of Berberine in the Regulation of CYP3A4 and P-gp via PXR Signaling Pathway

目的:研究黄连素对CYP3A4和P-gp的调控是否通过孕烷X受体(PXR)途径。方法:利用PXR慢病毒干扰质粒pLKO. 1-PXR转染包装细胞293T制备重组慢病毒,感染人肝癌细胞HepG2。根据pLKO. 1载体特性,采用嘌呤霉素筛选得到稳定干扰PXR表达的HepG2细胞。采用实时荧光定量PCR法和Western Blot法检测经由黄连素处理的HepG2细胞和PXR基因沉默细胞的CYP3A4及P-gp的mRNA水平和蛋白表达。结果:细胞转染质粒后,其PXR蛋白表达均显著下降(P <0. 01),但CYP3A4和P-gp的蛋白表达差异无统计学意义(P> 0. 05)。黄连素对PXR基因沉默细胞的CYP3A4、P-gp蛋白和mRNA表达的抑制效应较HepG2细胞显著减弱(P <0. 05或P <0. 01)。结论:黄连素可通过PXR信号通路调控CYP3A4和P-gp的表达,但并非唯一途径。

Objective： To investigate the regulation of CYP3A4 and P-gp by berberine hydrochloride（ BBR） via pregnane X receptor（ PXR） pathway. Methods： pLKO. 1-PXR vector,a lentivirus plasmid expressing PXR shRNA,was packaged into 293 T cells.Human hepatoma（ HepG2） cells were infected with the lentivirus and the cell clones stably expressing PXR shRNA were selected by puromycin according to pLKO. 1 vector characteristics. Real-time RT-PCR and Western blot were used to evaluate CYP3A4 and P-gp mRNA and protein in berberine treated HepG2 cells and PXR-silenced HepG2 cells. Results： The PXR expression in PXR silenced cells significantly decreased（ P〈0. 01） when compared with that in HepG2 cells,while there was no significant difference（ P〉0. 05） in the expression of CYP3A4 and P-gp between the groups. Compared with that in HepG2 cells,the inhibition of berberine on the mRNA and protein expression of CYP3A4 and P-gp in PXR-silenced HepG2 cells was weakened（ P〈0. 05 or P〈0. 01）. Conclusion： Berberine can regulate the expression of CYP3A4 and P-gp via PXR signaling pathway,while it is not the only one.