Effects of luteolin on the activities of cytochrome P450 enzymes in rats by Cocktail probe approach [Cocktail探针药物法评价木犀草素对大鼠CYP450酶活性影响]

作者：Gao L.; Song K.-X.; Ren J.-P.; Yi J.; Li Q.; Tang J.-L.^*

来源：中国新药杂志, 2022, 31(6): 590-595.

• Cocktail probe approach
• CYP450
• Liver microsomes
• Luteolin
• Probe drugs

Objective: To investigate the potential inhibition of luteoline against CYP1A2, CYP3A4 and CYP2C9 activities in rat liver microsomes through cocktail approach in vitro. Methods: Incubation method was developed and validated for in vitro CYP450 inhibition assay, comprising rat liver microsomes, a cocktail of substrates, luteoline or positive inhibitors. The experiment was divided into blank control group, luteolin group and positive control group. After incubation, the formation of metabolites was investigated by HPLC. The IC50 values of luteolin and positive inhibitors were calculated by GraphPad Prism 9.0. Results: The optimal concentration of rat liver microsomes was 1.5 mg?mL-1, and incubation time was 30 min. The Km values of phenacetin, testosterone and tolbutamide were 85.62, 146.90 and 78.36 μmol?L-1, respectively. The established incubation system was validated by known positive inhibitors. The IC50 values of luteolin to CYP1A2, CYP3A4, and CYP2C9 were 19.92, 4.64 and 9.29 μmol?L-1, respectively. Conclusion: An improved cocktail approach was established and validated for CYP inhibition assay. Luteolin has inhibitory activities against CYP1A2, CYP3A4 and CYP2C9.