Sinha Indu, Modesto Jennifer, Krebs Nicolle M, Stanley Anne E, Walter Vonn A, Richie John P, Muscat Joshua E, Sinha Raghu
Department of Biochemistry and Molecular Biology, Penn State Cancer Institute, Hershey, United States.
Department of Public Health Sciences, Penn State Cancer Institute, Hershey, United States.
Tob Induc Dis. 2021 Jun 29;19:56. doi: 10.18332/tid/138336. eCollection 2021.
Smoking is the leading cause of preventable disease. Although smoking results in an acute effect of relaxation and positive mood through dopamine release, smoking is thought to increase stress symptoms such as heart rate and blood pressure from nicotine-induced effects on the HPA axis and increased cortisol. Despite the importance in understanding the mechanisms in smoking maintenance, little is known about the overall protein and physiological response to smoking. There may be multiple functions involved that if identified might help in improving methods for behavioral and pharmacological interventions. Therefore, our goal for this pilot study was to identify proteins in the saliva that change in response to an acute smoking event versus acute sham smoking event in smokers and non-smokers, respectively.
We employed the iTRAQ technique followed by Mass Spectrometry to identify differentially expressed proteins in saliva of smokers and non-smokers after smoking cigarettes and sham smoking, respectively. We also validated some of the salivary proteins by ELISA or western blotting. In addition, salivary cortisol and salivary amylase (sAA) activity were measured.
In all, 484 salivary proteins were identified. Several proteins were elevated as well as decreased in smokers compared to non-smokers. Among these were proteins associated with stress response including fibrinogen alpha, cystatin A and sAA. Our investigation also highlights methodological considerations in study design, sampling and iTRAQ analysis.
We suggest further investigation of other differentially expressed proteins in this study including ACBP, A2ML1, APOA4, BPIB1, BPIA2, CAH1, CAH6, CYTA, DSG1, EST1, GRP78, GSTO1, sAA, SAP, STAT, TCO1, and TGM3 that might assist in improving methods for behavioral and pharmacological interventions for smokers.
吸烟是可预防疾病的主要原因。尽管吸烟通过多巴胺释放产生放松和积极情绪的急性效应，但吸烟被认为会增加压力症状，如因尼古丁对下丘脑 - 垂体 - 肾上腺（HPA）轴的影响以及皮质醇增加导致的心率和血压升高。尽管了解吸烟维持机制很重要，但对于吸烟的整体蛋白质和生理反应知之甚少。可能涉及多种功能，如果能够识别这些功能，可能有助于改进行为和药物干预方法。因此，我们这项初步研究的目标是分别确定吸烟者和非吸烟者在急性吸烟事件与急性假吸烟事件后唾液中发生变化的蛋白质。
我们采用同位素标记相对和绝对定量（iTRAQ）技术，随后进行质谱分析，以分别鉴定吸烟者和非吸烟者在吸烟和假吸烟后唾液中差异表达的蛋白质。我们还通过酶联免疫吸附测定（ELISA）或蛋白质免疫印迹法验证了一些唾液蛋白质。此外，还测量了唾液皮质醇和唾液淀粉酶（sAA）活性。
总共鉴定出484种唾液蛋白质。与非吸烟者相比，吸烟者中有几种蛋白质升高以及降低。其中包括与应激反应相关的蛋白质，如纤维蛋白原α、胱抑素A和sAA。我们的研究还强调了研究设计、采样和iTRAQ分析中的方法学考虑因素。
我们建议对本研究中其他差异表达的蛋白质进行进一步研究，包括酰基辅酶A结合蛋白（ACBP）、α2巨球蛋白样蛋白1（A2ML1）、载脂蛋白A4（APOA4）、杀菌/渗透增强蛋白B1（BPIB1）、杀菌/渗透增强蛋白A2（BPIA2）、碳酸酐酶1（CAH1）、碳酸酐酶6（CAH6）、胱抑素A（CYTA）、桥粒芯糖蛋白1（DSG1）、酯酶1（EST1）、葡萄糖调节蛋白78（GRP78）、谷胱甘肽S-转移酶O1（GSTO1）、sAA、血清淀粉样蛋白P成分（SAP）、信号转导和转录激活因子（STAT）、跨膜蛋白154（TCO1）和转谷氨酰胺酶3（TGM3），这些蛋白质可能有助于改进吸烟者的行为和药物干预方法。
