Nicotine Component of Cigarette Smoke Extract (CSE) Decreases the Cytotoxicity of CSE in BEAS-2B Cells Stably Expressing Human Cytochrome P450 2A13.
Nicotine Component of Cigarette Smoke Extract (CSE) Decreases the Cytotoxicity of CSE in BEAS-2B Cells Stably Expressing Human Cytochrome P450 2A13.
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香烟烟雾提取物 (CSE) 的尼古丁成分可降低稳定表达人细胞色素 P450 2A13 的 BEAS-2B 细胞中 CSE 的细胞毒性
DOI：
10.3390/ijerph14101221
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发表时间：
2017-10-13
影响因子：
--
通讯作者：
Wang SL
中科院分区：
文献类型：
--
作者：
Ji M;Zhang Y;Li N;Wang C;Xia R;Zhang Z;Wang SL
细胞色素P450 2A13是一种主要在人体呼吸系统表达的肝外酶，已有研究报道其参与了香烟烟雾的代谢和毒性作用。我们先前发现尼古丁通过抑制细胞色素P4-2A13代谢来抑制烟碱(4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone，NNK)的代谢，但其对香烟烟雾其他成分的影响尚不清楚。将卷烟烟气提取物(CSE)分解为尼古丁部分(CSE-N)和非尼古丁部分(CSE-O)，采用高效液相色谱(HPLC)和超高效液相色谱串联质谱(UPLC-MS/MS)对其进行分离、纯化和鉴定。分别用细胞计数试剂盒8(CCK-8)和流式细胞仪检测稳定表达细胞色素P450 2A13(B-2A13)和载体(B-V)的永生化人支气管上皮细胞(BEAS-2B)的细胞活力和细胞凋亡率。有趣的是，CSE和CSE-O对BEAS-2B细胞有毒性，而CSE-N的细胞毒性较小。CSE-O对B-2A13细胞的毒性大于对B-V细胞的毒性(IC50分别为2.49%和7.06%)，后者被CYP抑制剂8-甲氧补骨脂素(8-MOP)抑制。CSE-O而不是CSE或CSE-N促进了B-2A13细胞的凋亡，而不是B-V细胞。相应地，与CSE-N和CSE相比，CSE-O显著改变了B-2A13细胞中三对促凋亡和抗凋亡蛋白的表达，即Bcl2相关X蛋白/B细胞淋巴瘤-2(Bax/Bcl2)、裂解聚(二磷酸腺苷-核糖)聚合酶(C-PARP/PARP)和C-caspase-3/caspase-3。此外，CSE-N和CSE-O的重组(CSE-O/N)表现出与原始CSE相似的细胞毒性和细胞凋亡。这些结果表明，尼古丁成分降低了CYP2A13对CSE的代谢激活，有助于理解CYP2A13在吸烟引起的人类呼吸系统疾病中的关键作用。
Cytochrome P450 2A13 (CYP2A13), an extrahepatic enzyme mainly expressed in the human respiratory system, has been reported to mediate the metabolism and toxicity of cigarette smoke. We previously found that nicotine inhibited 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) metabolism by CYP2A13, but its influence on other components of cigarette smoke remains unclear. The nicotine component of cigarette smoke extract (CSE) was separated, purified, and identified using high-performance liquid chromatography (HPLC) and ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS), splitting CSE into a nicotine section (CSE-N) and nicotine-free section (CSE-O). Cell viability and apoptosis by Cell Counting Kit-8 (CCK-8) and flow cytometry assays were conducted on immortalized human bronchial epithelial (BEAS-2B) cells stably expressing CYP2A13 (B-2A13) or vector (B-V), respectively. Interestingly, CSE and CSE-O were toxic to BEAS-2B cells whereas CSE-N showed less cytotoxicity. CSE-O was more toxic to B-2A13 cells than to B-V cells (IC50 of 2.49% vs. 7.06%), which was flatted by 8-methoxypsoralen (8-MOP), a CYP inhibitor. CSE-O rather than CSE or CSE-N increased apoptosis of B-2A13 cells rather than B-V cells. Accordingly, compared to CSE-N and CSE, CSE-O significantly changed the expression of three pairs of pro- and anti-apoptotic proteins, Bcl-2 Associated X Protein/B cell lymphoma-2 (Bax/Bcl-2), Cleaved Poly (Adenosine Diphosphate-Ribose) Polymerase/Poly (Adenosine Diphosphate-Ribose) Polymerase (C-PARP/PARP), and C-caspase-3/caspase-3, in B-2A13 cells. In addition, recombination of CSE-N and CSE-O (CSE-O/N) showed similar cytotoxicity and apoptosis to the original CSE. These results demonstrate that the nicotine component decreases the metabolic activation of CYP2A13 to CSE and aids in understanding the critical role of CYP2A13 in human respiratory diseases caused by cigarette smoking.
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影响因子：
4.1
作者：
Shimada T;Murayama N;Yamazaki H;Tanaka K;Takenaka S;Komori M;Kim D;Guengerich FP
通讯作者：
Guengerich FP
影响因子：
3.1
作者：
Comer DM;Elborn JS;Ennis M
通讯作者：
Ennis M
DOI：
10.1016/s0140-6736(15)00340-2
发表时间：
2015-10-10
期刊：
Lancet (London, England)
影响因子：
--
作者：
Chen Z;Peto R;Zhou M;Iona A;Smith M;Yang L;Guo Y;Chen Y;Bian Z;Lancaster G;Sherliker P;Pang S;Wang H;Su H;Wu M;Wu X;Chen J;Collins R;Li L;China Kadoorie Biobank (CKB) collaborative group
通讯作者：
China Kadoorie Biobank (CKB) collaborative group
影响因子：
2.9
作者：
Chen, TA;Yang, FS;Chan, SO
通讯作者：
Chan, SO
影响因子：
4.7
作者：
Li, Lei;Megaraj, Vandana;Ding, Xinxin
通讯作者：
Ding, Xinxin