DNA甲基转移酶活性对NNK α-亚甲基羟基化代谢的影响  

Effects of activity of DNA methyltransferase on α-methylene hydroxylation metabolism of NNK

作　　者：王荣浩[1] 李琛[1] 毛健[1] 刘帅东 王丁众[1] 张启东[1] 霍现宽[1] 张建勋[1] WANG Rongha LI Chen MAO Jian LIU Shuaidong WANG Dingzhong ZHANG Qidong HUO Xiankuan ZHANG Jianxun(Zhengzhou Tobacco Research Institute of CNTC, Zhengzhou 450001, China Technology Center of China Tobacco Shaanxi Industrial Co., Ltd., Baoji 721013, Shaanxi, China)

机构地区：[1]中国烟草总公司郑州烟草研究院 [2]陕西中烟工业有限责任公司技术中心

出　　处：《烟草科技》2017年第10期42-47,共6页Tobacco Science & Technology

基　　金：国家自然科学基金面上项目"改性介孔材料固相萃取/UPLC-MSn方法研究烟草特有亚硝胺在动物体内代谢"(21175153);国家自然科学基金青年基金项目"烟碱在大鼠脑内代谢的活体动态分析及其神经递质化学效应的实时评价"(21307163)

摘　　要：为准确评价DNA甲基转移酶(DNMT)的活性对4-(甲基亚硝氨基)-1-(3-吡啶基)-1-丁酮(NNK)通过α-亚甲基羟基化代谢途径产生O^6-甲基鸟嘌呤(O^6-mG)的影响,采用超高效液相色谱-高分辨质谱(UPLC-HRMS)方法,分别对100 mg/kg NNK染毒后的生理盐水处理组、地西他滨处理组小鼠肝脏和肺中DNMT的活性、DNA甲基化程度,以及4-(3-吡啶基)-4-氧代丁酸(OPBA)、4-(3-吡啶基)-4-羟基丁酸(HPBA)和O^6-mG的质量分数进行了测定。结果表明:(1)地西他滨能够抑制DNA甲基转移酶活性,降低DNA甲基化水平。(2)地西他滨对NNK通过α-亚甲基羟基化代谢产生OPBA和HPBA的途径无显著影响。(3)抑制DNA甲基转移酶的活性,能减少NNK通过α-亚甲基羟基化代谢产生O^6-mG的量。In order to accurately evaluate the activity of DNA methyltransferase（DNMT） on the amount of O^6-methylguanine（O^6-m G） produced by α-methylene hydroxylation metabolism of 4-（methylnitrosamino）-1-（3-pyridyl）-1-butanone（NNK）, ultra-high performance liquid chromatography-high resolution mass spectrometry（UPLC-HRMS） was used to determine the activity of DNMT, the level of DNA methylation, the contents of 4-oxo-4-（3-pyridyl）butyric acid（OPBA）, 4-hydroxy-4-（3-pyridyl）butyric acid（HPBA） and O^6-m G in the livers and lungs of mice in physiological saline-treated group and decitabine-treated group after 100 mg/kg NNK administration separately. The results showed that： 1） Decitabine inhibited the activity of DNA methyltransferase and decreased the level of DNA methylation. 2） Decitabine did not significantly affect the contents of OPBA and HPBA produced by α-methylene hydroxylation metabolism of NNK. 3） The amount of O^6-m G produced through α-methylene hydroxylation metabolism of NNK could be reduced by inhibiting the activity of DNA methyltransferase.

关 键 词：DNA甲基转移酶(DNMT) DNA甲基化 4-(甲基亚硝氨基)-1-(3-吡啶基)-1-丁酮(NNK) 地西他滨 O^6-甲基鸟嘌呤(O^6-mG) 超高效液相色谱-高分辨质谱联用(UPLC-HRMS) 

分 类 号：TS416[农业科学—烟草工业]