Abstract
INTRODUCTIONSmoking 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.METHODSWe 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.RESULTSIn 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.CONCLUSIONSWe 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.
Highlights
Smoking is the leading cause of preventable disease
The study was conducted in accordance with the Declaration of Helsinki, and the protocol was approved by the Institutional Review Board (IRB# PRAMS042265EP) of Penn State University College of Medicine
RESULTS isobaric tags for relative and absolute quantitation (iTRAQ) analysis A total of 484 proteins were identified in the pooled saliva samples by iTRAQ analysis (Supplementary file Table S1)
Summary
Smoking is the leading cause of preventable disease. 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. A major reason smokers have a difficult time quitting is the need to alleviate psychological stress and negative mood This is known as selfmedication, where smoking is used to lower negative mood and stress, and increase positive mood. The stress hormone cortisol is the standard biomarker of stress response, representing the activation of the hypothalamic–pituitary–adrenal (HPA) axis pathway. Both salivary amylase protein levels and sAA activity have been used in stress research. Given the limited effectiveness of standard of care treatment for nicotine dependence (e.g. counseling and nicotine pharmacotherapy), a greater understanding of the role of stress could potentially lead to new and improved methods for behavioral and pharmacological interventions
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