Acetaminophen UDP-glucuronosyltransferase in ferrets: species and gender differences, and sequence analysis of ferret UGT1A6.
Acetaminophen UDP-glucuronosyltransferase in ferrets: species and gender differences, and sequence analysis of ferret UGT1A6.
复制标题
雪貂中的对乙酰氨基酚 UDP-葡萄糖醛酸基转移酶：物种和性别差异以及雪貂 UGT1A6 的序列分析。
DOI：
10.1046/j.1365-2885.2001.00366.x
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发表时间：
2001
影响因子：
1.3
通讯作者：
Court,MH
中科院分区：
文献类型：
--
作者：
Court,MH
本研究的主要目的是确定雪貂的葡萄糖醛酸对乙酰氨基酚是否比其他物种更慢，如果是的话，研究这种差异的分子基础。用8只雪貂、4人、4只猫、4只狗、大鼠、小鼠、牛、马、猴、猪和兔的肝微粒体测定对乙酰氨基酚- UDP -葡萄糖醛酸转移酶（UGT）的活性。采用酶动力学分析方法探讨了雄性和雌性雪貂肝脏的性别差异。使用UGT特异性抗体对微粒体蛋白进行免疫印迹。最后，对对乙酰氨基酚- UGT的主要基因UGT1A6外显子1区进行了测序。对乙酰氨基酚的葡萄糖醛酸化在雪貂的肝脏中相对较慢，除了猫。性别差异也很明显，雄性雪貂肝脏的内在清除率（Vmax/Km）值明显高于雌性雪貂肝脏。此外，vmax值与UGT1A亚家族特异性抗体鉴定的两个蛋白带的密度测量相关。在雪貂UGT1A6基因的外显子1或侧翼区域未发现有害突变。综上所述，雪貂和猫一样，肝脏对乙酰氨基酚的葡萄糖醛酸代谢相对较慢。然而，与猫的UGT1A6由假基因编码和功能失调不同，雪貂的UGT1A6基因没有缺陷，这可以解释低活性的原因。
The principal objective of this study was to determine whether ferrets glucuronidate acetaminophen more slowly compared with other species, and if so investigate the molecular basis for the difference. Acetaminophen‐UDP‐glucuronosyltransferase (UGT) activities were measured using hepatic microsomes from eight ferrets, four humans, four cats, four dogs, rat, mouse, cow, horse, monkey, pig and rabbit. Gender differences between male and female ferret livers were explored using enzyme kinetic analysis. Immunoblotting of microsomal proteins was also performed using UGT‐specific antibodies. Finally, the exon 1 region of UGT1A6, a major acetaminophen‐UGT, was sequenced. Glucuronidation of acetaminophen was relatively slow in ferret livers compared with livers from all other species except cat. Gender differences were also apparent, with intrinsic clearance (Vmax/Km) values significantly higher in male compared with female ferret livers. Furthermore,Vmaxvalues correlated with densitometric measurements of two protein bands identified with a UGT1A subfamily‐specific antibody. No deleterious mutations were identified in the exon 1 or flanking regions of the ferret UGT1A6 gene. In conclusion, like cats, ferret livers glucuronidate acetaminophen relatively slowly. However, unlike cats, in which UGT1A6 is encoded by a pseudogene and dysfunctional, there are no defects in the ferret UGT1A6 gene which could account for the low activity.