Strassburg C P, Nguyen N, Manns M P, Tukey R H
Department of Pharmacology and the Cancer Center, University of California, San Diego, La Jolla, California 92093-0636, USA.
Gastroenterology. 1999 Jan;116(1):149-60. doi: 10.1016/s0016-5085(99)70239-8.
BACKGROUND & AIMS: The contribution of glucuronidation toward human drug metabolism is carried out by the Super gene family of UDP-glucuronosyltransferases (UGTs). Regulation of the human UGT1A locus is tissue specific, resulting in the unique expression of multiple hepatic and extrahepatic gene products. Studies were undertaken to examine UGT1A expression in human hepatic and colonic tissues.
UGT1A messenger RNA, protein, catalytic activity, and substrate kinetics were studied in 5 samples of normal hepatic and sigmoid colon tissue using duplex reverse-transcription polymerase chain reaction (RT-PCR), enzymatic and Western blot analysis, and indirect immunofluorescence analysis.
Specific patterns of UGT1A gene expression occur in the liver and colon, which were consistent with different banding patterns as detected by Western blot analysis using a UGT1A-specific antibody. However, microsomal UGT activities in colon were up to 96-fold lower for many phenolic substrates, a finding that was not concordant with RT-PCR and Western blot analysis. Interestingly, UGT activity toward tertiary amines and some steroid hormones was equal.
Differences of glucuronidation activity between human liver and colon suggest that UGT1A activity may be regulated as a result of the relative presence of individual isoforms with differing catalytic activities or by tissue-specific modulators after gene expression.
葡萄糖醛酸化对人体药物代谢的作用由尿苷二磷酸葡萄糖醛酸基转移酶(UGTs)超基因家族完成。人类UGT1A基因座的调控具有组织特异性,导致多种肝内和肝外基因产物的独特表达。本研究旨在检测UGT1A在人肝脏和结肠组织中的表达情况。
采用双重逆转录聚合酶链反应(RT-PCR)、酶法、蛋白质免疫印迹分析及间接免疫荧光分析,对5份正常肝脏和乙状结肠组织样本中的UGT1A信使核糖核酸、蛋白质、催化活性及底物动力学进行研究。
UGT1A基因在肝脏和结肠中呈现特定的表达模式,这与使用UGT1A特异性抗体进行蛋白质免疫印迹分析所检测到的不同条带模式一致。然而,结肠微粒体对许多酚类底物的UGT活性比肝脏低96倍,这一结果与RT-PCR和蛋白质免疫印迹分析不一致。有趣的是,结肠对叔胺和某些甾体激素的UGT活性与肝脏相当。
人肝脏和结肠中葡萄糖醛酸化活性的差异表明,UGT1A活性可能是由于具有不同催化活性的个体同工型的相对存在,或基因表达后组织特异性调节剂的作用而受到调控。