西格列汀对阿托伐他汀细胞内药物蓄积的影响及其改变OATP1B1的表达  

Effect of sitagliptin on drug accumulation of intracellular atorvastatin and changing the expression of OATP1B1

作　　者：金海英[1] 麦一峰[2] 叶小磊 黄静[1] 余莉[1] 林薇[1] JIN Haiying;MAI Yifeng;YE Xiaolei;HUANG Jing;YU Li;LIN Wei(Department of Pharmacy, The Affiliated Hospital of Medicine College, Ningbo University, Ningbo 315020, China;Department of Cardiology, The Affiliated Hospital of Medicine College, Ningbo University, Ningbo 315020, China;Institute of Medical Science, Ningbo University, Ningbo 315020, China)

机构地区：[1]宁波大学医学院附属医院药剂科,宁波315020 [2]宁波大学医学院附属医院心内科,宁波315020 [3]宁波大学医学科学研究所,宁波315020

出　　处：《中国临床药学杂志》2019年第4期258-263,共6页Chinese Journal of Clinical Pharmacy

基　　金：浙江省宁波大学医学院资助项目(编号XYY16044)

摘　　要：目的证实阿托伐他汀可以通过OATP1B1进行转运,探讨西格列汀对阿托伐他汀在HEK293T细胞内药物蓄积的影响,为两药在临床的联合应用提供实验依据。方法建立OATP1B1过表达HEK293T(HEK293T^(1B1))细胞。采用MTT法检测不同浓度的阿托伐他汀和西格列汀对HEK293T细胞活力的影响;LC-MS/MS法检测阿托伐他汀单药和/或与西格列汀混合后在HEK293T^(1B1)细胞内不同时间点的浓度;免疫印迹法分析西格列汀对OATP1B1表达水平的影响。结果阿托伐他汀在20和40μmol·L^(-1)的高浓度下对HEK293T细胞有略微的抑制作用;HEK293T^(1B1)细胞对阿托伐他汀的摄取量随着孵育时间的延长相应增加,但在30~60 min时摄取量几乎不再变化。两药合用时,随着加入的西格列汀浓度的升高,阿托伐他汀在HEK293T^(1B1)细胞中的蓄积量进一步增加,差异具有统计学意义(P<0.05);免疫印迹实验的结果显示1.0μmol·L^(-1)的西格列汀对OATP1B1蛋白表达水平的上调作用最明显,在10 min时可达到高峰,20 min以后,蛋白的表达量逐渐减少。结论阿托伐他汀在肝细胞的转运水平与OATP1B1表达水平有关,而西格列汀可上调OATP1B1表达,因此两药具有潜在相互作用的可能。AIM To confirm that atorvastatin could be transported through OATP1 B1, investigate the effect of sitagliptin on the drug accumulation of atorvastatin in HEK293 T cells, and provide the experimental basis for the combination of the two drugs in clinical application. METHODS OATP1 B1 over expression HEK293 T(HEK293 T1 B1) cells model was established. MTT assay was used to determine the effect of different concentrations of atorvastatin and sitagliptin on the viability of HEK293 T cells. The concentration of atorvastatin alone or mixture of sitagliptin in the model cells at different time points was detected by liquid chromatography-tandem mass spectrometry(LC-MS/MS). The concentration of atorvastatin after mixed with different concentration of sitagliptin was detected with the same method. The effect of sitagliptin on the expression level of OATP1 B1 was analyzed by Western blotting. RESULTS Atorvastatin had a slight inhibitory effect on HEK293 T cells at the concentration of 20 and 40 μmol·L-1. Uptake of atorvastatin by HEK293 T1 B1cells increased with the extension of incubation time, but almost unchanged at 30-60 min. In the combined group, the concentration of atorvastatin in HEK293 T1 B1cells was further increased with the increase of the concentration of sitagliptin, and the difference was statistically significant(P<0.05). The results of the Western blot showed that the expression level of OATP1 B1 protein was significantly increased by the sitagliptin with 1.0 μmol·L-1,which reached peaks at 10 min and gradually decreased after 20 min. CONCLUSION Atorvastatin transport level in hepatocytes is related to the expression of OATP1 B1, while sitagliptin can up-regulate the expression of OATP1 B1, so there is a potential interaction between the two drugs.

关 键 词：西格列汀 阿托伐他汀 OATP1B1 LC-MS/MS 药物蓄积 药物相互作用 

分 类 号：R965[医药卫生—药理学]