肝脏p450还原酶特异性敲除小鼠模型在毒理学中的应用

肝脏肝脏 P450 还原酶特异性敲除小鼠模型在毒理学中的应用还原酶特异性敲除小鼠模型在毒理学中的应用 1 Application of Liver-specific Cytochrome P450 Reductase knockout mice in Toxicology 肖瑛，武元峰，薛翔，顾军，任进* Xiao Ying, Wu Yuanfeng, Xue Xiang, Gu Jun, Ren Jin 中国科学院上海药物研究所，新药研究国家重点实验室,上海 201203 State Key Laboratory for New Drug Research，Shanghai Institute of Materia Medica,Chinese Academy of Sciences, Shanghai, 201203 关键词关键词: NADPH-细胞色素 P450 酶还原酶，基因敲除，器官毒性 Key Word: NADPH-P450 Cytochrome Reductase, Knockout, Organ Toxicity 1.引言引言 微粒体细胞色素 P450 单加氧酶（P450，CYP）在内源与外源性化合物的生 物转化中起着重要的作用。P450 酶蛋白种类的多样性及其底物的重叠性使 P450 酶系可以催化多种类型的反应，不仅对许多外来物质如杀虫剂及环境有毒 物质具有代谢作用，还参与一些起重要生理功能的内源性物质如激素、脂肪酸 的代谢。P450 酶的功能与生物学作用的重要性，使其研究具有广阔的应用前景。 通过调控其表达和活性，可望改变生物体内外源性化合物尤其是药物的代谢， 对于药物的研究和开发也有重要的意义。由于 P450 酶系的生物学重要性，自其 发现以来，该酶系的研究一直是药理学和毒理学中一个十分引人注目的领域。 P450 酶基因的多态性使其有多种亚型且多具有类似的功能和相似的组织分 布特征，使在一个特异组织中很难去区别单个 P450 酶亚型的功能。目前已有多 个 P450 酶亚型敲除（knockout，KO）小鼠模型，这些模型已被广泛用来研究 特异性 P450 酶亚型在外源性化合物的代谢和毒性中的作用1。然而这些 KO 1 基金项目： 基金项目：国家重点基础研究发展计划 973 项目（编号：2006CB504700） 作者简介：作者简介：肖瑛（1981-） ，女，博士研究生，研究方向为药物毒理学。 *通讯作者：通讯作者：任进，Email: CDSER_SIMMmail. Tel 83(6):818-28. 2.Okada H, Watanabe Y, Inoue T, et al. Transgene-derived hepatocyte growth factor attenuates reactive renal fibrosis in aristolochic acid nephrotoxicity. J Nephrol Dial Transplant 2003;18(12):2515-23. 3.Gu J, Weng Y, Zhang QY, et al. Liver-specific deletion of the NADPH- cytochrome P450 reductase gene: impact on plasma cholesterol homeostasis and the function and regulation of microsomal cytochrome P450 and heme oxygenase. J J Biol Chem 2003;278(28):25895-901. 4.Weng Y, Fang C, Turesky RJ, et al. Determination of the role of target tissue metabolism in lung carcinogenesis using conditional cytochrome P450 reductase-null mice. J Cancer Res 2007;67(16):7825-32. 5.Fang C, Gu J, Xie F, et al. Deletion of the NADPH-cytochrome P450 reductase gene in cardiomyocytes does not protect mice against doxorubicin-mediated acute cardiac toxicity. J Drug Metab Dispos 2008;36(8):1722-8. 6.Gu J, Chen CS, Wei Y, et al. A mouse model with liver-specific deletion and global suppression of the NADPH-Cytochrome P450 reductase gene: Characterization and utility for in vivo studies of cyclophosphamide disposition. J J Pharmacol Exp Ther 2007;321(1):9-17. 7.Henderson CJ, Otto DM, Carrie D, et al. Inactivation of the hepatic cytochrome P450 system by conditional deletion of hepatic cytochrome P450 reductase. J J Biol Chem 2003;278(15):13480-6. 8.Gu J, Cui H, Behr M, et al. In vivo mechanisms of tissue-selective drug toxicity: effects of liver-specific knockout of the NADPH-cytochrome P450 reductase gene on acetaminophen toxicity in kidney, lung, and nasal mucosa. J Mol Pharmacol 2005;67(3):623-30. 9.Pass GJ, Carrie D, Boylan M, et al. Role of hepatic cytochrome p450s in the pharmacokinetics and toxicity of cyclophosphamide: studies with the hepatic cytochrome p450 reductase null mouse. J Cancer Res 2005;65(10):4211-7. 10. Fang C, Behr M, Xie F, et al. Mechanism of chloroform-induced renal toxicity: non-involvement of hepatic cytochrome P450-dependent metabolism. J Toxicol Appl Pharmacol 2008;227(1):48-55. 11. Xiao Y, Ge M, Xue X, et al. Hepatic cytochrome P450s metabolize aristolochic acid and reduce its kidney toxicity. J Kidney Int 2008;73(11):1231-9. 12. Arlt VM, Stiborova M, Henderson CJ, et al. Metabolic activation of benzoapyrene in vitro by hepatic cytochrome P450 contrasts with detoxification in vivo: experiments with hepatic cytochrome P450 reductase null mice. J Carcinogenesis 2008;29(3):656-65. 13. Arlt VM, Henderson CJ, Wolf CR, et al. Bioactivation of 3-aminobenzanthrone, a human metabolite of the environmental pollutant 3-nitrobenzanthrone: evidence for DNA adduct formation mediated by cytochrome P450 enzymes and peroxidases. J Cancer Lett 2006;234(2):220-31. 14. Henderson CJ, Pass GJ, Wolf CR. The hepatic cytochrome P450 reductase null mouse as a tool to identify a successful candidate entity. J Toxicol Lett 2006;162(1):111-7. 15. Weng Y, DiRusso CC, Reilly AA, et al. Hepatic gene expression changes in mouse models with liver-specific deletion or global suppression of the NADPH- cytochrome P450 reductase gene. Mechanistic implications for the regulation of microsomal cytochrome P450 and the fatty liver p