cAMP-PKA信号通路对L02细胞药物代谢酶CYP3A的调控作用

目的 探讨cAMP-PKA信号通路对细胞色素P4503A(CYP3A)在正常肝细胞L02表达中的作用.方法 102细胞中分别加入8-溴-环腺苷酸(8-Br-cAMP)10μmol·L<'-1>及抑制剂H-8910μmol·L<'-1>,培养48 h,分别采用完整细胞免疫组化法、Western印迹法和红霉素-N-脱甲基法...

Regulation of cAMP-PKA singal pathway on cytochrome P4503A in L02 cells

OBJECTIVE To observe the effect of cAMP-PKA signal pathway on the activity of cytochrome P4503A (CYP3A) in L02 cells, and to explore the mechanism. METHODS L02 cells were treated with agonist 8-Br-cAMP 10 μmol·L^-1 and inhibitor H-89 10 μmol·L^-1. The expression of pregnane X receptor (PXR) and protein kinase A (PKA) protein were detected by Western blotting and immunohistochemistry. The activity of CYP3A was detected by ultraviolet spectrophotometry. RESULTS ]Compared with the activity of CYP3A 〔(0.28±0.02)μmol·min^-1·g^-1 protein〕, the expression of PXR (0.45±0.14) and PKA (211±20) in normal control group, the activity of CYP3A〔(0.43±0.01)μmol·min^-1·g^-1 protein〕, and the expression of PXR (0.69±0.21) and PKA (296±18) increased in 8-Br-cAMP group while the expression of PKA (176±14) decreased. The activity of CYP3A 〔(0.39±0.05)μmol·min^-1·g^-1 protein〕 and the expression of PXR (0.47±0.13) were no statistical significance in H-89 group. CONCLUSIONThe cAMP- PKA singal pathway can regulate the CYP3A in L02 cells.