琥珀酸曲格列汀对db/db小鼠CYP450酶和药物转运体的影响

Effects of Trelagliptin Succinate on CYP450 Enzymes and Drug Transporters in db/db Mice

目的探讨琥珀酸曲格列汀对db/db小鼠肝组织中2种主要的CYP450酶(CYP3A4、CYP2D6)和3种药物转运体(GLUT2、MRP2、NTCP)的表达水平的影响。方法将18只db/db小鼠随机分成模型对照组、琥珀酸曲格列汀组、磷酸西格列汀组,6只C57小鼠为正常对照组。琥珀酸曲格列汀组每周灌胃给予琥珀酸曲格列汀17.29 mg·kg^(-1),磷酸西格列汀组灌胃给予磷酸西格列汀16.71 mg·kg^(-1)·d^(-1),连续给药8周。苏木精-伊红(HE)染色观察琥珀酸曲格列汀对db/db小鼠肝组织病理学是否有改善作用;酶联免疫吸附测定实验(ELISA)法测定各组糖化血红蛋白水平以评价db/db小鼠的糖化血红蛋白是否相对于C57健康小鼠升高,并探讨琥珀酸曲格列汀能否降低糖化血红蛋白;采用实时荧光定量多聚核苷酸链式反应(RT-PCR)和Western blotting法检测各小鼠肝组织中CYP3A4、CYP2D6、GLUT2、MRP2和NTCP的mRNA水平和蛋白表达情况。结果与模型对照组比较,琥珀酸曲格列汀组肝组织病理状态改善,肝细胞空泡变性减轻;CYP3A4 mRNA表达水平无明显差异(P>0.05),而CYP3A4蛋白表达水平显著升高(P<0.01);CYP2D6 mRNA和蛋白表达水平无明显差异(P>0.05);GLUT2 mRNA和蛋白表达水平显著升高(P<0.01);MRP2 mRNA和蛋白表达水平无明显差异(P>0.05);NTCP mRNA表达水平无明显差异(P>0.05),而NTCP蛋白表达水平显著升高(P<0.01)。结论琥珀酸曲格列汀可增加db/db小鼠的CYP3A4、GLUT2和NTCP的表达,而对CYP2D6和MRP2的表达无影响。

Objective To explore the effects of trelagliptin succinate on the expression levels of 2 major CYP450 enzymes(CYP3A4,CYP2D6)and three drug transporters(GLUT2,MRP2,NTCP)in liver tissues of the db/db mice.Methods Eighteen db/db mice were randomly divided into model control group,trelagliptin succinate group,sitagliptin phosphate group,and six C57 mice were set as a normal control group.The trelagliptin succinate group was intragastrically administered with trelagliptin succinate 17.29 mg·kg^(-1)·w^(-1),and the sitagliptin phosphate group was intragastrically administered with sitagliptin phosphate 16.71 mg·kg^(-1)·d^(-1) for eight weeks.HE staining was used to observe whether trelagliptin succinate could improve liver histopathology in db/db mice;ELISA assay was used to measure the glycated hemoglobin levels in each group to evaluate whether the glycated hemoglobin of db/db mice was higher than that of C57 healthy mice.The mRNA and protein expression levels of CYP3A4,CYP2D6,GLUT2,MRP2,and NTCP in the liver tissues of each mouse were detected by Real-time PCR and Western blotting.Results Compared with the model control group,trelagliptin succinate could improve the pathological state of liver tissue,and attenuate vacuolar degeneration of hepatocytes;there was no significant difference in the expression level of CYP3A4 mRNA(P>0.05),while the protein expression level of CYP3A4 was significantly increased(P<0.01);CYP2D6 mRNA and protein expression levels had no significant difference(P>0.05);GLUT2 mRNA and protein expression levels were significantly increased(P<0.01);MRP2 mRNA and protein expression levels had no significant difference(P>0.05);NTCP showed no significant difference in mRNA expression level(P>0.05),while NTCP protein expression level was significantly increased(P<0.01).Conclusion Trelagliptin succinate can increase the expressions of CYP3A4,GLUT2 and NTCP,but does not affect on the expressions of CYP2D6 and MRP2.