唐草片与依非韦伦相互作用及对CYP450酶的调控研究
发布时间：2018-07-29 19:20
【摘要】：目的：艾滋病(AIDS)患者常用唐草片进行辅助甚至替代治疗,然而中药成分复杂,其对抗病毒药物的影响及其作用机制尚不清楚。为了减少潜在的药物与药物相互作用,临床上推荐服用依非韦伦(EFV)1小时后再服唐草片。然而这种服用方式影响患者的依次性,本研究探讨唐草片与EFV同时服用时,唐草片对EFV药代动力学的影响及作用机制；且在此基础上深入研究唐草片对CYP450酶的影响。方法：本研究首先研究不同给药方式下唐草片对依非韦伦(EFV)药动学特性的影响,将18只SD大鼠随机分为三组：单独服用EFV、EFV和唐草片同时服用、以及服用EFV 60分钟后服唐草片。用色谱串联质谱(LC-MS/MS)法检测EFV的血药浓度。然后采用定量PCR技术研究唐草片对EFV代谢酶CYP450酶CYP1A1、CYP1B1、CYP2B1、CYP2B2、CYP2E1和CYP3A1的调控。此外,我们采用差速离心法分离肝微粒体,SDS-PAGE分离其蛋白质,切取CYP450所在的三条电泳带,用液相色谱串联质谱进行鉴定。对鉴定的蛋白质进行生物信息学分析,Expasy blast搜索分析CYP2C11的特征多肽及限定片断离子的m.z800,选择CYP2C11中m.z为711.5的多肽进行MRM定量分析,该多肽的离子对为：711.5→232.1、711.5→319.2、711.5→466.2和711.5→595.3；内标(IS)ESAT-6肽的离子对为：735.5→215.3、735.5→389.3、735.5→460.3和735.5→524.3。采用相对峰(分析物／内标)面积进行相对定量分析。结果：我们研究了唐草片对EFV药代动力学的影响及作用机制。具体从以下二方面展开：1) EFV以及唐草片单次给药大鼠模型的药代动力学研究显示：EFV单独服用组、EFV和唐草片同时服用以及服用EFV 60分钟后服唐草片组中EFV的曲线下面积(AUC0-∞)分别为17.97、10.06和14.32 mg.1.h。统计学分析显示：与EFV单独服用相比,EFV和唐草片同时服用时的曲线下面积(AUC0-∞)、半衰期(T1.2)以及保留时间(MRT)显著下调(p0.05)。当EFV和唐草片同时服用时,唐草片显著减低EFV的生物利用度和血浆半衰期,而在服用EFV60分钟后服用唐草片,两者的AUC0-∞接近,没有统计学差异。2) 对EFV代谢相关代谢酶的定量PCR分析结果显示：唐草片不影响EFV的6个CYP450代谢酶的mRNA水平。进一步的蛋白质组研究在肝微粒体中鉴定16个CYP450同工酶。为了分析唐草片对CYP450酶的影响,采用基于质谱的多反应监测技术(MRM)进行定量分析。发现：与EFV单独给药组相比,在EFV与唐草片合并用药组中CYP2C11上调了1.6倍。结论：唐草片与EFV同时服用时,降低EFV的生物利用度；体外酸性环境中唐草片提高EFV的溶解；唐草片不影响EFV直接相关代谢酶的表达,唐草片提高CYP2C11的蛋白质表达。但唐草片对EFV调控的具体机制有待深入研究。
[Abstract]:Objective: in (AIDS) patients, Tangcao tablets are commonly used to assist or even substitute therapy. However, the effect of Chinese traditional medicine on the antiviral drugs and its mechanism are not clear. To reduce potential drug-drug interactions, it is recommended to take efeveron (EFV) 1 hour later before taking Tangcao tablets. However, the effect of Tangcao tablet on the pharmacokinetics of EFV and the mechanism of its action were investigated, and the effect of Tangcao tablet on CYP450 enzyme was also studied. Methods: the effect of Tangcao tablets on the pharmacokinetic characteristics of (EFV) was studied. Eighteen Sprague-Dawley rats were randomly divided into three groups: taking both EFV and Tangcao tablets alone and taking Tangcao tablets 60 minutes after taking EFV. The plasma concentration of EFV was determined by tandem mass spectrometry (LC-MS/MS). Then the quantitative PCR technique was used to study the regulation of EFV metabolizing enzyme CYP450 enzyme CYP1A1C CYP2B1C CYP2B1CY CYP2B2CY CYP2E1 and CYP3A1 in Tangcao tablets. In addition, SDS-PAGE was used to isolate the protein from liver microsomes by differential centrifugation. Three electrophoresis bands of CYP450 were cut and identified by liquid chromatography tandem mass spectrometry (LC-MS). Bioinformatics analysis was carried out on the identified protein. The characteristic polypeptide of CYP2C11 and the specific fragment ion of CYP2C11 were analyzed by blast search. The polypeptides with m 路z = 711.5 in CYP2C11 were selected for MRM quantitative analysis. The ion pairs of this peptide were: 711.5 / 232.1711.5 / 319.2711.5 / 466.2 and 711.5 / 595.3respectively. The ion pairs of the internal standard (IS) ESAT-6 peptide are: 1: 735.5, 215.3735.5, 389.3735.5, 460.3, and 735.5, 524.3. Relative peak (analyte / internal standard) area was used for relative quantitative analysis. Results: the effect of Tangcao tablet on the pharmacokinetics of EFV and its mechanism were studied. The pharmacokinetic study of EFV and Tangcao tablets in rat model showed that the EFV in the group treated with EFV alone and Tangcao tablets were taken at the same time and after taking EFV for 60 minutes. The area under line (AUC0- 鈭，
本文编号：2153730
[Abstract]:Objective: in (AIDS) patients, Tangcao tablets are commonly used to assist or even substitute therapy. However, the effect of Chinese traditional medicine on the antiviral drugs and its mechanism are not clear. To reduce potential drug-drug interactions, it is recommended to take efeveron (EFV) 1 hour later before taking Tangcao tablets. However, the effect of Tangcao tablet on the pharmacokinetics of EFV and the mechanism of its action were investigated, and the effect of Tangcao tablet on CYP450 enzyme was also studied. Methods: the effect of Tangcao tablets on the pharmacokinetic characteristics of (EFV) was studied. Eighteen Sprague-Dawley rats were randomly divided into three groups: taking both EFV and Tangcao tablets alone and taking Tangcao tablets 60 minutes after taking EFV. The plasma concentration of EFV was determined by tandem mass spectrometry (LC-MS/MS). Then the quantitative PCR technique was used to study the regulation of EFV metabolizing enzyme CYP450 enzyme CYP1A1C CYP2B1C CYP2B1CY CYP2B2CY CYP2E1 and CYP3A1 in Tangcao tablets. In addition, SDS-PAGE was used to isolate the protein from liver microsomes by differential centrifugation. Three electrophoresis bands of CYP450 were cut and identified by liquid chromatography tandem mass spectrometry (LC-MS). Bioinformatics analysis was carried out on the identified protein. The characteristic polypeptide of CYP2C11 and the specific fragment ion of CYP2C11 were analyzed by blast search. The polypeptides with m 路z = 711.5 in CYP2C11 were selected for MRM quantitative analysis. The ion pairs of this peptide were: 711.5 / 232.1711.5 / 319.2711.5 / 466.2 and 711.5 / 595.3respectively. The ion pairs of the internal standard (IS) ESAT-6 peptide are: 1: 735.5, 215.3735.5, 389.3735.5, 460.3, and 735.5, 524.3. Relative peak (analyte / internal standard) area was used for relative quantitative analysis. Results: the effect of Tangcao tablet on the pharmacokinetics of EFV and its mechanism were studied. The pharmacokinetic study of EFV and Tangcao tablets in rat model showed that the EFV in the group treated with EFV alone and Tangcao tablets were taken at the same time and after taking EFV for 60 minutes. The area under line (AUC0- 鈭，
本文编号：2153730
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