J Toxicol Sci. 2010 Aug;35(4):571-6. doi: 10.2131/jts.35.571.
The nuclear receptor superfamily consists of ligand-dependent transcription factors. Among them, constitutive androstane receptor (CAR) plays a key role in the detoxification of xenobiotics, inducing various drug-metabolizing enzymes including human CYP2B6 and its homologues of other species. AMP-activated protein kinase (AMPK) acts as an important energy sensor, being activated by an increased AMP/ATP ratio. CAR is activated by phenobarbital (PB) treatment. It has been recently reported that AMPK is involved in PB-mediated CYP2B induction both in vitro and in vivo. We investigated the relationship between the functions of AMPK and CAR in rat primary hepatocyte. The AMPK-activator 5-aminoimidazole-4-Carboxamide-1-beta-Ribofuranoside (AICAR) unexpectedly repressed PB-induced CYP2B mRNA expression as well as AMPK-inhibitor compound C. In contrast, both the AMPK-activator metformin and the constitutive active form of AMPK enhanced PB-induced PB-responsive enhancer module-driven reporter gene expression. We demonstrated that AICAR prevented nuclear translocation of CAR in an AMPK-independent manner in rat primary hepatocytes. AICAR might be a convenient probe for studying the mechanisms of PB-induced activation, especially nuclear translocation, of CAR in rat primary hepatocytes.
核受体超家族由配体依赖性转录因子组成。其中,组成型雄烷受体(CAR)在解毒外来物方面发挥关键作用,诱导各种药物代谢酶,包括人 CYP2B6 及其在其他物种中的同源物。AMP 激活的蛋白激酶(AMPK)作为一种重要的能量传感器,通过增加 AMP/ATP 比值而被激活。CAR 被苯巴比妥(PB)处理激活。最近有报道称,AMPK 参与了体外和体内 PB 介导的 CYP2B 诱导。我们研究了 AMPK 和 CAR 在大鼠原代肝细胞中的功能关系。出乎意料的是,AMPK 激活剂 5-氨基咪唑-4-羧酰胺-1-β-D-呋喃核糖苷(AICAR)抑制了 PB 诱导的 CYP2B mRNA 表达以及 AMPK 抑制剂化合物 C。相比之下,AMPK 激活剂二甲双胍和 AMPK 的组成性激活形式均增强了 PB 诱导的 PB 反应增强子模块驱动的报告基因表达。我们证明 AICAR 以 AMPK 独立的方式阻止了 CAR 在大鼠原代肝细胞中的核易位。AICAR 可能是研究 PB 诱导的 CAR 激活,特别是核易位机制的便捷探针,特别是在大鼠原代肝细胞中。