Effects of Saikosaponin D on CYP1A2 and CYP2D6 in HepaRG Cells

Abstract

Background

Bupleurum is one of the most important traditional Chinese medicines and an ingredient in many compound preparations. It is widely used together with other drugs in clinical practice, and thus there is great potential for drug–drug interactions. Saikosaponin D (SsD) is a major bioactive triterpenoid saponin extracted from Bupleurum with anti-inflammatory, anticancer, antioxidative, and antihepatic fibrosis effects. Effects of the main components of Bupleurum on cytochromes P450 (CYPs) need to be clarified in the clinical application of combination therapies of formulations containing SsD or Bupleurum.

Purpose

This study aimed to investigate the effects of SsD on the CYP1A2 and CYP2D6 mRNAs, protein expression, and relative enzyme activities in HepaRG cells.

Methods

HepaRG cells were cultured with SsD at concentrations of 0.5, 1, 5 and 10 μM for 72 hours. mRNA and protein expression of CYP1A2 and CYP2D6 were analyzed with real-time PCR and Western blot analysis. Relative enzyme activities were analyzed with HPLC based on consumption of the specific probe substrate.

Results

SsD significantly induced expression of mRNA and increased relative activity of CYP1A2 in HepaRG cells after the cells had been treated with SsD at concentrations of 1, 5 and 10 μM. SsD also induced protein expression of CYP1A2 at concentrations of 5 and 10 μM. SsD exhibited an inductive effect on CYP2D6 mRNA and protein expression, while increasing the relative activity of CYP2D6 at concentrations of 5 and 10 μM.

Conclusion

This study is the first to investigate the effect of SsD on CYP1A2 and CYP2D6 in HepaRG cells, and the results may provide some useful information on potential drug–drug interactions related to clinical preparations containing SsD or Bupleurum.

Keywords:

• saikosaponin D
• HepaRG cells
• drug–drug interactions

Acknowledgments

This work was financially supported by grants from the National Natural Science Foundation of China (31460246), Guizhou Provincial Science and Technology Foundation (Qiankehe J [2013] 2323), Project of Special Funds for Science and Technology Cooperation in Guizhou Province and Zunyi City (Shengshikehe [2015] 53), Collaborative Innovation Center of Guizhou Traditional Chinese Medicine and Ethnic Medicine (Qianjiaokeyanfa [2012] 311).

Disclosure

The authors declare that they have no conflicts of interest in this work.