Correlation between Male Infertility and Oxidative Stress
Oxidative stress has been implicated in the etiology of male infertility. Identifying and understanding the proteins that play a key role in oxidative stress-induced pathophysiology of male infertility is important as it can help in the development of some of these key proteins as potential biomarkers.
ROS levels were measured in seminal ejaculates by chemiluminescence assay using luminol (5-amino-2, 3- dihydro-1,4-phthalazinedione; Sigma, St Louis, MO). Test samples consisted of luminol (10 μL, 5 mM) and 400 μL of semen. Negative controls were prepared by replacing the sperm suspension with 400 μL phosphate buffered saline. Positive control included 400 μL of PBS and 50 μL of hydrogen peroxide (30%; 8.8 M) in triplicate. Chemiluminescence was measured for 15 min using a Berthold luminometer (Autolumat Plus LB 953).
Samples were subjected to LC-MS/MS analysis through in-solution digestion of proteins for peptide characterization. The expression profile of proteins present in human spermatozoa was examined using proteomic and bioinformatic analysis to elucidate the regulatory pathways of oxidative stress.
Proteins have been identified that help protect against oxidative stress and are uniquely present in the seminal plasma of the ROS- men. Men exhibiting high levels of ROS in their seminal ejaculate are likely to exhibit proteins that are either downregulated or oxidatively modified, and these could potentially contribute to male infertility.
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