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, r2 = 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.

中文翻译：

---

非酒精性脂肪性肝病和糖尿病与人肝中CYP3A4蛋白表达和活性的降低有关。

非酒精性脂肪肝疾病（NAFLD）是西方人群慢性肝病的主要原因。我们调查了非酒精性脂肪肝疾病（NAFLD）和糖尿病对脑死亡供体（n = 74）人肝组织中CYP3A4活性的影响。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）两类。咪达唑仑向其1-羟基代谢产物的转化率用于评估人肝微粒体（HLM）的体外CYP3A4活性。蛋白质组学方法用于定量CYP3A4和相关酶的蛋白质表达。此外，建立了基于生理学的药代动力学（PBPK）模型，可以预测NAFL和NASH肝脏中咪达唑仑的浓度。CYP3A4活性在NAFL和NASH中分别比正常供者低1.9倍和3.1倍（p <0.05）。NAFL和NASH供体的内在清除率（CLint）分别低2.7-（p <0.05）和4.1倍（p <0.01）。CYP3A4蛋白表达在NAFL和NASH供体中显着降低（p <0.05），并在多元线性回归分析中说明了咪达唑仑的显着羟化变异性（β= 0.869，r2 = 0.762，P <0.01）。糖尿病也与CYP3A4活性和蛋白表达降低有关。咪达唑仑CLint和CYP3A4蛋白的丰度均随着肝脂肪积累的增加而显着降低。年龄和性别与观察到的变化没有任何显着相关性。预计NAFL和NASH的咪达唑仑暴露分别高1.7和2.3倍，在这些疾病状态下可能会导致更长的镇静时间。数据表明，NAFLD和糖尿病与肝CYP3A4活性降低有关。因此，有必要进一步评估这些发现对CYP3A4底物的功效和安全性的临床后果。