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.



中文翻译：

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柴胡皂苷 D 对 HepaRG 细胞 CYP1A2 和 CYP2D6 的影响

背景： 柴胡是重要的中药材之一，也是许多复方制剂的成分。它在临床实践中广泛与其他药物一起使用，因此具有很大的药物相互作用潜力。Saikosaponin D (SsD) 是一种从柴胡中提取的主要生物活性三萜皂苷，具有抗炎、抗癌、抗氧化和抗肝纤维化作用。柴胡主要成分对细胞色素P450（CYPs）的影响在含SsD或柴胡制剂联合治疗的临床应用中需要明确。
目的：本研究旨在探讨 SsD 对 HepaRG 细胞中 CYP1A2 和 CYP2D6 mRNA、蛋白表达和相关酶活性的影响。
方法： HepaRG 细胞与浓度为 0.5、1、5 和 10 μM 的 SsD 一起培养 72 小时。用实时荧光定量PCR和蛋白质印迹分析分析CYP1A2和CYP2D6的mRNA和蛋白表达。基于特定探针底物的消耗用HPLC分析相对酶活性。
结果：在用浓度为 1、5 和 10 μM 的 SsD 处理细胞后，SsD 显着诱导 HepaRG 细胞中 mRNA 的表达并增加 CYP1A2 的相对活性。SsD 还在 5 和 10 μM 浓度下诱导 CYP1A2 的蛋白质表达。SsD 表现出对 CYP2D6 mRNA 和蛋白质表达的诱导作用，同时在 5 和 10 μM 浓度下增加 CYP2D6 的相对活性。
结论：本研究首次研究了 SsD 对 HepaRG 细胞中 CYP1A2 和 CYP2D6 的影响，该结果可能为与含有 SsD 或柴胡的临床制剂相关的潜在药物-药物相互作用提供一些有用的信息。