靛玉红衍生物PHⅡ-7通过PXR-CYP3A5/ABCB1信号通路对他克莫司转运及代谢的调节及其作用机制研究  

Regulation of indirubin derivative PHⅡ-7 on tacrolimus transport and metabolism via PXR-CYP3A5/ABCB1 signaling pathway and its mechanism

作　　者：刘红 景艳 邹德琴[2] 侯雄军[3] 彭洪薇 周健[2] 黎玉华[2] 熊冬生[4] 温金华[2] 魏筱华[1,2] LIU Hong;JING Yan;ZOU De-qin;HOU Xiong-jun;PENG Hong-wei;ZHOU Jian;LI Yu-hua;XIONG Dong-sheng;WEN Jin-hua;WEI Xiao-hua(School of Pharmacy,Nanchang University;Dept of Pharmacy,the First Affiliated Hospital of Nanchang University;Jiangxi Provincial People's Hospital,Nanchang 330006,China;Institute of Hematology,Chinese Academy of Medical Sciences,Tianjin 300020,China)

机构地区：[1]南昌大学药学院 [2]南昌大学第一附属医院药学部 [3]江西省人民医院药学部,江西南昌330006 [4]中国医学科学院血液学研究所,天津300020

出　　处：《中国药理学通报》2020年第7期927-934,共8页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金项目(No.81860035);吴阶平医学基金会项目(No.320.6750.19090-36)。

摘　　要：目的探究靛玉红衍生物PHⅡ-7通过激活PXR-CYP3A5/ABCB1信号通路进而影响他克莫司(FK506)转运与代谢的机制。方法本研究以五味子甲素(Sch-A)为阳性对照,大鼠体内实验研究PHⅡ-7对FK506药动学的影响;MTT法检测PHⅡ-7、Sch-A对HepG2和LS174T细胞的IC50值;RT-qPCR和Western blot检测PHⅡ-7、Sch-A对PXR/CYP3A4/3A5/ABCB1信号通路转录和蛋白水平的影响。结果与Sch-A组比较,PHⅡ-7组AUC0-t、CL明显增加(P<0.05);HepG2和LS174T细胞PHⅡ-7的IC50分别为(17.82±1.33)μmol·L^-1和(7.17±0.37)μmol·L^-1,Sch-A的IC50分别为(40.38±4.1)μmol·L^-1和(44.1±1.94)μmol·L^-1;HepG2和LS174T细胞经PHⅡ-7和Sch-A处理后,PXR、CYP3A4/3A5和ABCB1的mRNA和蛋白表达均明显降低(P<0.05),且呈剂量与时间依赖性。结论 PHⅡ-7可通过PXR-CYP3A5/ABCB1信号通路调节FK506的转运与代谢,进而增强FK506的体内疗效。Aim To investigate the mechanism of the regulatory effect of indirubin derivative PHⅡ-7 on the transport and metabolism of FK506 by activating the PXR-CYP3A5/ABCB1 signaling network.Methods In vivo experiments were used to investigate the effect of PHⅡ-7 on the pharmacokinetics of FK506 in rats(set Sch-A as positive control).Additionally,IC 50 values of PHⅡ-7 and Sch-A on HepG2 and LS174T cells were determined by MTT assay.Effects of PHⅡ-7 and Sch-A on mRNA and protein of PXR/CYP3A4/3A5/ABCB1 gene were detected by RT-qPCR and Western blot.Results Compared with Sch-A,PHⅡ-7 significantly increased AUC 0-t,CL of FK506(P<0.05).IC 50 of PHⅡ-7 on HepG2 and LS174T cells were(17.82±1.33)μmol·L^-1 and(7.17±0.37)μmol·L^-1,while IC 50 of Sch-A on HepG2 and LS174T cells were(40.38±4.1)μmol·L^-1 and(44.1±1.94)μmol·L^-1,respectively.HepG2 and LS174T cells were treated with PHⅡ-7 or Sch-A,and the mRNA and protein expressions of PXR,CYP3A4/3A5 and ABCB1 genes markedly decreased in a dose-and time-dependent manner(P<0.05).Conclusions In vivo and in vitro experiments indicate that PHⅡ-7 could regulate the transport and metabolism of FK506 through PXR-CYP3A5/ABCB1 signaling pathway,and strengthen the FK506 curative effect further.

关 键 词：PHⅡ-7 他克莫司 五味子甲素 转运 代谢 PXR-CYP3A5/ABCB1信号通路 

分 类 号：R-332[医药卫生] R284.1