隐丹参酮对细胞色素P450酶CYP3A4和CYP2C9的影响

Effects of Cryptotanshinone on CYP3A4 and CYP2C9 expressions

目的研究丹参中脂溶性成分隐丹参酮对Chang肝脏细胞细胞色素P450酶(CYP450)CYP3A4、CYP2C9表达的影响,明确丹参主要成分对CYP450酶的作用,以阐明基于CYP450酶隐丹参酮与其他药物间相互作用的分子基础,为临床合理使用含隐丹参酮的丹参制剂提供依据。方法采用CCK-8法考察隐丹参酮对Chang肝脏细胞活力的影响,以确定药物的给药剂量范围;通过Western bloting法考察隐丹参酮在给药后1、3和7 d对Chang肝脏细胞中CYP3A4和CYP2C9蛋白表达的影响;通过定量R T-PCR法考察隐丹参酮对Chang肝脏细胞中CYP3A4和CYP2C9 mRNA表达的影响。结果给药1 d,隐丹参酮对CYP3A4蛋白及mRNA表达无显著影响。但随时间延长,给药3 d和7 d,隐丹参酮可抑制CYP3A4蛋白及mRNA的表达(P<0.05)。给药1、3和7 d,隐丹参酮对CYP2C9蛋白及mRNA的表达均无显著影响。结论连续给予隐丹参酮可抑制CYP3A4 mRNA和蛋白的表达。提示临床在连续使用含隐丹参酮时,需要关注其与CYP3A4底物药物之间可能的相互作用。

【Objective】 To investigate the effects of Cryptotanshinone on CYP3A4 and CYP2C9 expressions in Chang liver cells, confirm its effect on cytochrome P450 enzyme and analyze the drug-drug interaction so as to lay a foundation for rational medicine administration. 【Methods】 CCK-8 assay was performed to determine the proliferation of Chang liver cells after treatment with Cryptotanshinone for 1 day, 3 and 7 days.Quantificational reverse transcription polymerase chain reaction and Western blot were performed to determine mRNA and protein expressions of CYP3A4 and CYP2C9 respectively. 【Results】 After 1-day treatment, Cryptotanshinone had no effect on protein or mRNA expression of CYP3A4. However, Cryptotanshinone significantly inhibited the mRNA and protein expressions of CYP3A4（P 0.05） after 3- and 7-day treatment, and had no significant effect on the mRNA and protein expressions of CYP2C9 after 1-, 3- or 7-day treatment.【Conclusions】 Continual administration of Cryptotanshinone could inhibit the protein and mRNA expressions of CYP3A4, which suggests that the drug-drug interaction should be concerned about in clinic.