Biomedical and Pharmaceutical Sciences, College of Pharmacy , University of Rhode Island , Kingston , Rhode Island 02881 , United States.
Departments of Medicine, Pathology, Neurology, and Neurosurgery , Rhode Island Hospital and the Warren Alpert Medical School of Brown University , Providence , Rhode Island 02903 , United States.
Mol Pharm. 2018 Jul 2;15(7):2621-2632. doi: 10.1021/acs.molpharmaceut.8b00159. Epub 2018 Jun 11.
Nonalcoholic fatty liver disease (NAFLD) is a major cause of chronic liver disease in the Western population. We investigated the association of nonalcoholic fatty liver disease (NAFLD) and diabetes mellitus on CYP3A4 activity in human liver tissue from brain dead donors ( n = 74). Histopathologically graded livers were grouped into normal ( n = 24), nonalcoholic fatty liver (NAFL, n = 26), and nonalcoholic steatohepatitis (NASH, n = 24) categories. The rate of conversion of midazolam to its 1-hydroxy metabolite was used to assess in vitro CYP3A4 activity in human liver microsomes (HLM). A proteomics approach was utilized to quantify the protein expression of CYP3A4 and related enzymes. Moreover, a physiologically based pharmacokinetic (PBPK) model was developed to allow prediction of midazolam concentration in NAFL and NASH livers. CYP3A4 activity in NAFL and NASH was 1.9- and 3.1-fold ( p < 0.05) lower than normal donors, respectively. Intrinsic clearance (CLint) was 2.7- ( p < 0.05) and 4.1-fold ( p < 0.01) lower in donors with NAFL and NASH, respectively. CYP3A4 protein expression was significantly lower in NAFL and NASH donors ( p < 0.05) and accounted for significant midazolam hydroxylation variability in a multiple linear regression analysis (β = 0.869, r = 0.762, P < 0.01). Diabetes was also associated with decreased CYP3A4 activity and protein expression. Both midazolam CLint and CYP3A4 protein abundance decreased significantly with increase in hepatic fat accumulation. Age and gender did not exhibit any significant association with the observed alterations. Predicted midazolam exposure was 1.7- and 2.3-fold higher for NAFL and NASH, respectively, which may result in a longer period of sedation in these disease-states. Data suggests that NAFLD and diabetes are associated with the decreased hepatic CYP3A4 activity. Thus, further evaluation of clinical consequences of these findings on the efficacy and safety of CYP3A4 substrates is warranted.
非酒精性脂肪性肝病 (NAFLD) 是西方人群慢性肝病的主要病因。我们研究了脑死亡供体的人肝组织中非酒精性脂肪性肝病 (NAFLD) 和糖尿病对 CYP3A4 活性的影响 (n = 74)。组织病理学分级的肝脏分为正常 (n = 24)、非酒精性脂肪肝 (NAFL，n = 26) 和非酒精性脂肪性肝炎 (NASH，n = 24) 类别。米唑仑转化为 1-羟基代谢物的速率用于评估人肝微粒体 (HLM) 中的体外 CYP3A4 活性。采用蛋白质组学方法定量 CYP3A4 及其相关酶的蛋白表达。此外，开发了一种基于生理学的药代动力学 (PBPK) 模型，以允许预测米唑仑在 NAFL 和 NASH 肝脏中的浓度。NAFL 和 NASH 中的 CYP3A4 活性分别比正常供体低 1.9 倍和 3.1 倍 (p < 0.05)。内在清除率 (CLint) 在 NAFL 和 NASH 供体中分别降低了 2.7- (p < 0.05) 和 4.1 倍 (p < 0.01)。NAFL 和 NASH 供体中 CYP3A4 蛋白表达明显降低 (p < 0.05)，并在多元线性回归分析中对米唑仑羟化的变异性有显著影响 (β = 0.869，r = 0.762，P < 0.01)。糖尿病也与 CYP3A4 活性和蛋白表达降低有关。米唑仑 CLint 和 CYP3A4 蛋白丰度均随肝脂肪堆积增加而显著降低。年龄和性别与观察到的变化无显著相关性。对于 NAFL 和 NASH，预测的米唑仑暴露分别增加了 1.7 倍和 2.3 倍，这可能导致这些疾病状态下的镇静时间延长。数据表明，NAFLD 和糖尿病与肝 CYP3A4 活性降低有关。因此，需要进一步评估这些发现对 CYP3A4 底物疗效和安全性的临床后果。
