Abstract

The autonomic nervous system (ANS) plays a crucial role both in acute and chronic psychological stress eliciting changes in many local and systemic physiological and biochemical processes. Salivary secretion is also regulated by ANS. In this study, we explored salivary proteome changes produced in thirty-eight University students by a test stress, which simulated an oral exam. Students underwent a relaxation phase followed by the stress test during which an electrocardiogram was recorded. To evaluate the effect of an olfactory stimulus, half of the students were exposed to a pleasant odor diffused in the room throughout the whole session. Saliva samples were collected after the relaxation phase (T0) and the stress test (T1). State anxiety was also evaluated at T0 and T1. Salivary proteins were separated by two-dimensional electrophoresis, and patterns at different times were compared. Spots differentially expressed were trypsin digested and identified by mass spectrometry. Western blot analysis was used to validate proteomic results. Anxiety scores and heart rate changes indicated that the fake exam induced anxiety. Significant changes of α-amylase, polymeric immunoglobulin receptor (PIGR), and immunoglobulin α chain (IGHA) secretion were observed after the stress test was performed in the two conditions. Moreover, the presence of pleasant odor reduced the acute social stress affecting salivary proteome changes. Therefore, saliva proteomic analysis was a useful approach to evaluate the rapid responses associated to an acute stress test also highlighting known biomarkers.

Highlights

  • Test anxiety is a form of social anxiety defined as “the set of phenomenological, physiological and behavioral responses that accompany concern about possible negative consequences or failure on an exam or similar evaluative situations” [1]

  • We evaluated the pattern of salivary proteins secretion induced in thirty-eight students by a test anxiety condition, which simulated an oral exam

  • Since some essential oils (Eos), such as lavander, peppermint, orange, and bergamot have been reported to have anxiolytic effects [12,13], we investigated whether diffusion of an Eos odor was able to reduce the test anxiety response and modify related salivary proteomic changes

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Summary

Introduction

Test anxiety is a form of social anxiety defined as “the set of phenomenological, physiological and behavioral responses that accompany concern about possible negative consequences or failure on an exam or similar evaluative situations” [1]. It is one of the most frequent anxiety forms among students of both sexes and different ages [2,3,4,5]. Since some essential oils (Eos), such as lavander, peppermint, orange, and bergamot have been reported to have anxiolytic effects [12,13], we investigated whether diffusion of an Eos odor was able to reduce the test anxiety response and modify related salivary proteomic changes

Questionnaires and Heart Rate Results
Participants
Physiological Parameters
Experimental Protocol
Questionnaires and Heart Analysis
Salivary Samples
In-Gel Digestion and Mass Spectrometry
WB Analysis
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