Tissue-specific mRNA expression profiles of human phase I metabolizing enzymes except for cytochrome P450 and phase II metabolizing enzymes
Tissue-specific mRNA expression profiles of human phase I metabolizing enzymes except for cytochrome P450 and phase II metabolizing enzymes
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DOI：
10.2133/dmpk.21.357
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发表时间：
2006-01-01
影响因子：
2.1
通讯作者：
Naito, Shinsaku
中科院分区：
文献类型：
--
作者：
Nishimura, Masuhiro;Naito, Shinsaku
对52种人I相代谢酶中的每一种具有特异性的正向和反向引物对和TaqMan探针（醇脱氢酶、醛脱氢酶、醛氧化酶、二氢嘧啶脱氢酶、环氧化物水解酶、酯酶、含黄素单加氧酶、单胺氧化酶、前列腺素内过氧化物合酶、醌氧化还原酶和咕吨脱氢酶）和48种人II期代谢酶（乙酰转移酶、酰基-CoA：氨基酸N-酰基转移酶、UDP-葡糖醛酸基转移酶、谷胱甘肽S-转移酶、甲基转移酶和磺基转移酶）。在来自各种人体组织的单个和合并样本的总RNA中分析每种靶酶的mRNA表达水平（肾上腺、骨髓、脑、结肠、心脏、肾、肝、肺、胰腺、外周白细胞、胎盘、前列腺、唾液腺、骨骼肌、小肠、脊髓、脾、胃、睾丸、胸腺、甲状腺、气管、和子宫）中的DNA进行实时逆转录PCR。此外，还评价了肝脏中代表性人I相和II相代谢酶mRNA表达的个体差异。使用上述I相和II相代谢酶在23种不同人体组织中的mRNA表达谱来鉴定对这些酶表现出高转录活性的组织。这些结果有望在建立药物代谢介导的新化学实体筛选系统，在新药开发和有关疾病的临床诊断的研究是有价值的。
Pairs of forward and reverse primers and TaqMan probes specific to each of 52 human phase I metabolizing enzymes (alcohol dehydrogenase, aldehyde dehydrogenase, aldehyde oxidase, dihydropyrimidine dehydrogenase, epoxide hydrolase, esterase, flavin-containing monooxygenase, monoamine oxidase, prostaglandin endoperoxide synthase, quinone oxidoreductase, and xanthene dehydrogenase) and 48 human phase II metabolizing enzymes (acetyltransferase, acyl-CoA:amino acid N-acyltransferase, UDP-glucuronosyltransferase, glutathione S-transferase, methyltransferase, and sulfotransferase) were prepared. The mRNA expression level of each target enzyme was analyzed in total RNA from single and pooled specimens of various human tissues (adrenal gland, bone marrow, brain, colon, heart, kidney, liver, lung, pancreas, peripheral leukocytes, placenta, prostate, salivary gland, skeletal muscle, small intestine, spinal cord, spleen, stomach, testis, thymus, thyroid gland, trachea, and uterus) by real-time reverse transcription PCR using an ABI PRISM 7700 Sequence Detection System. Further, individual differences in the mRNA expression of representative human phase I and II metabolizing enzymes in the liver were also evaluated. The mRNA expression profiles of the above phase I and phase II metabolizing enzymes in 23 different human tissues were used to identify the tissues exhibiting high transcriptional activity for these enzymes. These results are expected to be valuable in establishing drug metabolism-mediated screening systems for new chemical entities in new drug development and in research concerning the clinical diagnosis of disease.