Abstract
Urine, reflecting many changes in the body, is a better source than blood for biomarker discovery. However, even under physiological conditions, the urine proteome often varies. Understanding how various regulating factors affect urine proteome helps link changes to urine proteome with urinary biomarkers of physiological conditions as well as corresponding diseases. To evaluate the possible impact of α1-adrenergic receptor on urine proteome, this study investigated effects of the specific inhibitor prazosin on the urine proteome in a rat model by using tandem mass tagging and two-dimensional liquid chromatography-tandem mass spectrometry. A total of 775 proteins were identified, approximately half of which were influenced by prazosin treatment, indicating that the sympathetic nervous system exerts a significant impact on urine proteome. Eight significantly changed proteins were previously annotated as urinary candidate biomarkers. Angiotensinogen, haptoglobin, and beta-2 microglobulin, which were reported to be associated with blood pressure, were validated via Western blot. Prazosin is widely used in clinical practice; thus, these protein changes should be considered when studying corresponding diseases such as hypertension, post-traumatic stress disorder and benign prostatic hyperplasia. The related physiological activities of α1-receptors, controlling blood pressure and fear response might significantly affect the urine proteome and warrant further biomarker studies.
Highlights
Biomarkers are the measurable changes associated with a physiological or pathophysiological process [1]
Elucidating how various regulating factors affect the urine proteome allows the association of changes to the urine proteome with urinary biomarkers of physiological conditions in PLOS ONE | DOI:10.1371/journal.pone
The urine proteome can be affected by anesthetics [3], which suggested that urinary proteins might be regulated by the central nervous system; changes in central nerve system are reflected in urine
Summary
Biomarkers are the measurable changes associated with a physiological or pathophysiological process [1]. Urinary protein identification was performed in six samples that before and after prazosin-treatment, and the protein changes were validated in the remaining twelve samples. The six urinary samples before and after the prazosin administration were individually labeled with 126, 127, 128, 129, 130 and 131 TMT reagents according to the manufacturer’s protocol (Thermo Fisher Scientific, Germany) and analyzed with two dimensional (2D) LC/MS/MS.
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