妊娠改变了治疗妊娠相关并发症的倾向和多种药物的暴露。先前的体外研究表明,妊娠相关激素(PRH)会改变人肝细胞中某些细胞色素P450(CYP)的表达和功能。然而,PRHs对非CYP药物代谢酶(DME)和转运蛋白肝脏浓度的影响在很大程度上仍然未知.在这项研究中,将来自五名女性供体的夹心培养的人肝细胞(SCHH)暴露于媒介物或PRH(雌酮,雌二醇,雌三醇,黄体酮,皮质醇,和胎盘生长激素),单独或联合给药,在一系列生理相关的PRH浓度持续72小时。使用定量靶向绝对蛋白质组学(QTAP)同位素稀释nanoLC-MS/MS方法,在SCHH膜部分中定量了33种肝非CYPDME和转运蛋白的绝对浓度。数据显示,PRH改变了各种DME和转运蛋白的绝对蛋白质浓度,异构体-,和肝细胞依赖的方式。总的来说,33种(24%)蛋白质中的8种在肝细胞供体中相对于载体对照(ANOVAp<0.05)表现出明显的PRH诱发的绝对蛋白质浓度净变化:1/11UGTs(9%;UGT1A4),4/6其他DME(67%;CES1、CES2、FMO5、POR),和3/16的转运蛋白(19%;OAT2,OCT3,P-GP)。另外8种(24%)蛋白质(UGT1A1、UGT2B4、UGT2B10、FMO3、OCT1、MRP2、MRP3、ENT1)在至少两个个体肝细胞供体中表现出绝对蛋白质浓度的显著PRH改变。相比之下,17(52%)蛋白质在肝细胞供体内或跨肝细胞供体的PRH没有表现出可辨别的影响。总的来说,这些结果为PRH对SCHH中非CYPDME和转运蛋白的影响提供了首次全面的定量蛋白质组学评估,并提供了对妊娠期间通过这些途径清除的药物底物配置改变的机制见解.
Pregnancy alters the disposition and exposure to multiple drugs indicated for pregnancy-related complications. Previous in vitro studies have shown that pregnancy-related hormones (PRHs) alter the expression and function of certain cytochrome P450s (CYPs) in human hepatocytes. However, the impact of PRHs on hepatic concentrations of non-CYP drug-metabolizing enzymes (DMEs) and transport proteins remain largely unknown. In this study, sandwich-cultured human hepatocytes (SCHH) from five female donors were exposed to vehicle or PRHs (estrone, estradiol, estriol, progesterone, cortisol, and placental growth hormone), administered individually or in combination, across a range of physiologically relevant PRH concentrations for 72 h. Absolute concentrations of 33 hepatic non-CYP DMEs and transport proteins were quantified in SCHH membrane fractions using a quantitative targeted absolute proteomics (QTAP) isotope dilution nanoLC-MS/MS method. The data revealed that PRHs altered the absolute protein concentration of various DMEs and transporters in a concentration-, isoform-, and hepatocyte donor-dependent manner. Overall, eight of 33 (24%) proteins exhibited a significant PRH-evoked net change in absolute protein concentration relative to vehicle control (ANOVA p < 0.05) across hepatocyte donors: 1/11 UGTs (9%; UGT1A4), 4/6 other DMEs (67%; CES1, CES2, FMO5, POR), and 3/16 transport proteins (19%; OAT2, OCT3, P-GP). An additional 8 (24%) proteins (UGT1A1, UGT2B4, UGT2B10, FMO3, OCT1, MRP2, MRP3, ENT1) exhibited significant PRH alterations in absolute protein concentration within at least two individual hepatocyte donors. In contrast, 17 (52%) proteins exhibited no discernable impact by PRHs either within or across hepatocyte donors. Collectively, these results provide the first comprehensive quantitative proteomic evaluation of PRH effects on non-CYP DMEs and transport proteins in SCHH and offer mechanistic insight into the altered disposition of drug substrates cleared by these pathways during pregnancy.