Khatri Raju, Kulick Natasha, Rementer Rebecca J B, Fallon John K, Sykes Craig, Schauer Amanda P, Malinen Melina M, Mosedale Merrie, Watkins Paul B, Kashuba Angela D M, Boggess Kim A, Smith Philip C, Brouwer Kim L R, Lee Craig R
Division of Pharmacotherapy and Experimental Therapeutics, UNC Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
Division of Pharmacoengineering and Molecular Pharmaceutics, UNC Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
J Pharm Sci. 2021 Jan;110(1):412-421. doi: 10.1016/j.xphs.2020.09.013. Epub 2020 Sep 12.
Pregnancy-related hormones (PRH) have emerged as key regulators of hepatic cytochrome P450 (CYP) enzyme expression and function. The impact of PRH on protein levels of CYP3A4 and other key CYP enzymes, and the metabolism of nifedipine (a CYP3A4 substrate commonly prescribed during pregnancy), was evaluated in primary human hepatocytes. Sandwich-cultured human hepatocytes (SCHH) from female donors were exposed to PRH (estradiol, estriol, estetrol, progesterone, and cortisol), individually or in combination as a cocktail. Absolute protein concentrations of twelve CYP isoforms in SCHH membrane fractions were quantified by nanoLC-MS/MS, and metabolism of nifedipine to dehydronifedipine in SCHH was evaluated. PRH significantly increased CYP3A4 protein concentrations and nifedipine metabolism to dehydronifedipine in a concentration-dependent manner. CYP3A4 mRNA levels in hepatocyte-derived exosomes positively correlated with CYP3A4 protein levels and dehydronifedipine formation in SCHH. PRH also increased CYP2B6, CYP2C8 and CYP2A6 levels. Our findings demonstrate that PRH increase nifedipine metabolism in SCHH by inducing CYP3A4 expression and alter expression of other key CYP proteins in an isoform-specific manner, and suggest that hepatocyte-derived exosomes warrant further investigation as biomarkers of hepatic CYP3A4 metabolism. Together, these results offer mechanistic insight into the increases in nifedipine metabolism and clearance observed in pregnant women.
妊娠相关激素（PRH）已成为肝细胞色素P450（CYP）酶表达和功能的关键调节因子。在原代人肝细胞中评估了PRH对CYP3A4和其他关键CYP酶蛋白水平以及硝苯地平（孕期常用的CYP3A4底物）代谢的影响。将来自女性供体的夹心培养人肝细胞（SCHH）单独或作为混合物暴露于PRH（雌二醇、雌三醇、雌四醇、孕酮和皮质醇）。通过nanoLC-MS/MS对SCHH膜组分中12种CYP同工型的绝对蛋白浓度进行定量，并评估SCHH中硝苯地平向脱氢硝苯地平的代谢。PRH以浓度依赖的方式显著增加CYP3A4蛋白浓度和硝苯地平向脱氢硝苯地平的代谢。肝细胞来源外泌体中的CYP3A4 mRNA水平与SCHH中的CYP3A4蛋白水平和脱氢硝苯地平形成呈正相关。PRH还增加了CYP2B6、CYP2C8和CYP2A6的水平。我们的研究结果表明，PRH通过诱导CYP3A4表达增加SCHH中硝苯地平的代谢，并以同工型特异性方式改变其他关键CYP蛋白的表达，提示肝细胞来源的外泌体作为肝脏CYP3A4代谢生物标志物值得进一步研究。总之，这些结果为孕妇中观察到的硝苯地平代谢和清除增加提供了机制性见解。
