烟碱、NAT和NAB对P450酶催化NNK和NNN代谢反应的抑制作用研究
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摘要
NNK和NNN是烟草特有的强致癌性成分,NNK和NNN只有在被P450酶代谢激活时才会产生致癌性。NNK、NNN及其它与之结构相似的化合物伴随烟气同时进入吸烟者体内,烟气中的其他成分烟碱、NAT和NAB能否对P450酶催化的NNK及NNN代谢反应产生抑制作用,值得研究。结果表明,P450 2A13酶催化NNK主要有OPB、HPB和OPBA三种代谢产物,催化NNN主要有HPB、Hydroxy acid和OPB三种代谢产物。上述酶促反应符合经典米氏动力学方程。烟碱、NAT和NAB对P450 2A13酶催化NNK和NNN的反应产生竞争性抑制作用,经计算三种化合物对不同代谢产物的抑制常数不同。三种抑制物对NNK代谢的抑制常数为:(1)OPB:8.51μM(烟碱)、0.21μM(NAT)、0.23μM(NAB);(2)HPB:25.01μM(烟碱)、0.71μM(NAT)、0.87μM(NAB);(3)OPBA:6.57μM(烟碱)、0.36μM(NAT)、0.50μM(NAB)。三种抑制物对NNN代谢抑制常数为:(1)HPB:0.98μM(烟碱)、1.37μM(NAT)、0.71μM(NAB);(2)Hydroxy acid:1.35μM(烟碱)、1.35μM(NAT)、1.01μM(NAB);(3)OPB:8.40μM(烟碱)、3.40μM(NAT)、3.04μM(NAB)。上述研究结果提示:烟气中有些结构相似的化合物尽管无致癌性或弱致癌性,却会对致癌物NNK和NNN的代谢激活产生抑制效应,而这种抑制效应在一定程度上可能会进一步影响NNK和NNN致癌作用。
引文
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18.Su,T.et al.Human cytochrome P450 CYP2A13:predominant expression in the respiratory tract and its high efficiency metabolic activation of a tobacco-specific carcinogen,4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone.Cancer Res 60,5074-5079(2000).
19.Megaraj,V.et al.Role of CYP2A13 in the bioactivation and lung tumorigenicity of the tobacco-specific lung procarcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone:in vivo studies using a CYP2A13-humanized mouse model.Carcinogenesis 35,131-137(2014).
20.Maser,E.,Richter,E.&Friebertsh?user,J.The Identification of 11β-hydroxysteroid Dehydrogenase as Carbonyl Reductase of the Tobacco-Specific Nitrosamine4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone.The FEBS Journal 238,484-489(1996).
21.Maser,E.,Friebertshauser,J.&Volker,B.Purification,characterization and NNK carbonyl reductase activities of 11beta-hydroxysteroid dehydrogenase type 1 from human liver:enzyme cooperativity and significance in the detoxification of a tobacco-derived carcinogen.Chem Biol Interact 143-144,435-448(2003).
22.Maser,E.Significance of reductases in the detoxification of the tobacco-specific carcinogen NNK.Trends Pharmacol Sci 25,235-237(2004).
23.Kramlinger,V.M.,von Weymarn,L.B.&Murphy,S.E.Inhibition and inactivation of cytochrome P450 2A6 and cytochrome P450 2A13 by menthofuran,beta-nicotyrine and menthol.Chem Biol Interact 197,87-92(2012).
24.Van Vleet,T.R.,Bombick,D.W.&Coulombe,R.A.,Jr.Inhibition of human cytochrome P450 2E1by nicotine,cotinine,and aqueous cigarette tar extract in vitro.Toxicol Sci 64,185-191(2001).
2.Hecht,S.S.&Hoffmann,D.Tobacco-specific nitrosamines,an important group of carcinogens in tobacco and tobacco smoke.Carcinogenesis 9,875-884(1988).
3.Rodgman,A.&Green,C.R.Toxic chemicals in cigarette mainstream smoke-hazard and hoopla.Beitr?ge zur Tabakforschung International 20,481-543(2003).
4.IARC,Volume 35(IARC,2012).
5.Jalas,J.R.,Hecht,S.S.&Murphy,S.E.Cytochrome P450 Enzymes as Catalysts of Metabolism of4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone,a Tobacco Specific Carcinogen.Chem.Res.Toxicol.18,95-110(2005).
6.Bao,Z.,He,X.Y.,Ding,X.,Prabhu,S.&Hong,J.Y.Metabolism of nicotine and cotinine by human cytochrome P450 2A13.Drug Metab Dispos 33,258-261(2005).
7.Denton,T.T.,Zhang,X.&Cashman,J.R.Nicotine-related alkaloids and metabolites as inhibitors of human cytochrome P-450 2A6.Biochemical Pharmacology 67,751-756(2004).
8.von Weymarn,L.B.,Brown,K.M.&Murphy,S.E.Inactivation of CYP2A6 and CYP2A13 during nicotine metabolism.J Pharmacol Exp Ther 316,295-303(2006).
9.Ordonez,P.et al.Nicotine,cotinine,and beta-nicotyrine inhibit NNK-induced DNA-strand break in the hepatic cell line HepaR G.Toxicol In Vitro 28,1329-1337(2014).
10.Jalas,J.R.,Ding,X.&Murphy,S.E.Comparative metabolism of the tobacco-specific introsamines4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone and 4-(methylnitrosamino)-1-butanol by rat cytochrome P450 2A3 and human cytochrome P450 2A13.Drug Metab.Dispos.31,1199-1202(2003).
11.Zhang,J.et al.Fully automated analysis of four tobacco-specific N-nitrosamines in mainstream cigarette smoke using two-dimensional online solid phase extraction combined with liquid chromatography-tandem mass spectrometry.Talanta 146,216-224(2016).
12.Patten,C.J.et al.Kinetic analysis of the activation of4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone by heterologously expressed human P450enzymes and the effect of P450-specific chemical inhibitors on this activation in human liver microsomes.Arch Biochem Biophys 333,127-138(1996).
13.Smith,T.J.et al.Metabolism of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone in human lung and liver microsomes and cytochromes P-450 expressed in hepatoma cells.Cancer Res 52,1757-1763(1992).
14.Smith,T.J.,Guo,Z.,Guengerich,F.P.&Yang,C.S.Metabolism of4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone(NNK)by human cytochrome P450 1A2 and its inhibition by phenethyl isothiocyanate.Carcinogenesis 17,809-813(1996).
15.Smith,T.J.,Stoner,G.D.&Yang,C.S.Activation of4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone(NNK)in human lung microsomes by cytochromes P450,lipoxygenase,and hydroperoxides.Cancer Res 55,5566-5573(1995).
16.Penman,B.W.et al.Characterization of a human cell line expressing high levels of c DNA-derived CYP2D6.Pharmacogenetics 3,28-39(1993).
17.Yamazaki,H.,Inui,Y.,Yun,C.-H.,Guengerich,F.P.&Shimada,T.Cytochrome P450 2E1 and 2A6enzymes as major catalysts for metabolic activation of N-nitrosodialkylamines and tobacco-related nitrosamines in human liver microsomes Carcinogenesis 13,1789-1794(1992).
18.Su,T.et al.Human cytochrome P450 CYP2A13:predominant expression in the respiratory tract and its high efficiency metabolic activation of a tobacco-specific carcinogen,4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone.Cancer Res 60,5074-5079(2000).
19.Megaraj,V.et al.Role of CYP2A13 in the bioactivation and lung tumorigenicity of the tobacco-specific lung procarcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone:in vivo studies using a CYP2A13-humanized mouse model.Carcinogenesis 35,131-137(2014).
20.Maser,E.,Richter,E.&Friebertsh?user,J.The Identification of 11β-hydroxysteroid Dehydrogenase as Carbonyl Reductase of the Tobacco-Specific Nitrosamine4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone.The FEBS Journal 238,484-489(1996).
21.Maser,E.,Friebertshauser,J.&Volker,B.Purification,characterization and NNK carbonyl reductase activities of 11beta-hydroxysteroid dehydrogenase type 1 from human liver:enzyme cooperativity and significance in the detoxification of a tobacco-derived carcinogen.Chem Biol Interact 143-144,435-448(2003).
22.Maser,E.Significance of reductases in the detoxification of the tobacco-specific carcinogen NNK.Trends Pharmacol Sci 25,235-237(2004).
23.Kramlinger,V.M.,von Weymarn,L.B.&Murphy,S.E.Inhibition and inactivation of cytochrome P450 2A6 and cytochrome P450 2A13 by menthofuran,beta-nicotyrine and menthol.Chem Biol Interact 197,87-92(2012).
24.Van Vleet,T.R.,Bombick,D.W.&Coulombe,R.A.,Jr.Inhibition of human cytochrome P450 2E1by nicotine,cotinine,and aqueous cigarette tar extract in vitro.Toxicol Sci 64,185-191(2001).