Cellular and Subcellular Oxidative Stress Parameters Following Severe Spinal Cord Injury

Authors

Nishant P. Visavadiya, University of KentuckyFollow
Samir P. Patel, University of Kentucky
Jenna L. VanRooyen, University of KentuckyFollow
Patrick G. Sullivan, University of KentuckyFollow
Alexander G. Rabchevsky, University of KentuckyFollow

Abstract

The present study undertook a comprehensive assessment of the acute biochemical oxidative stress parameters in both cellular and, notably, mitochondrial isolates following severe upper lumbar contusion spinal cord injury (SCI) in adult female Sprague Dawley rats. At 24 h post-injury, spinal cord tissue homogenate and mitochondrial fractions were isolated concurrently and assessed for glutathione (GSH) content and production of nitric oxide (NO^•), in addition to the presence of oxidative stress markers 3-nitrotyrosine (3-NT), protein carbonyl (PC), 4-hydroxynonenal (4-HNE) and lipid peroxidation (LPO). Moreover, we assessed production of superoxide (O₂^•-) and hydrogen peroxide (H₂O₂) in mitochondrial fractions. Quantitative biochemical analyses showed that compared to sham, SCI significantly lowered GSH content accompanied by increased NO^• production in both cellular and mitochondrial fractions. SCI also resulted in increased O₂^•- and H₂O₂ levels in mitochondrial fractions. Western blot analysis further showed that reactive oxygen/nitrogen species (ROS/RNS) mediated PC and 3-NT production were significantly higher in both fractions after SCI. Conversely, neither 4-HNE levels nor LPO formation were increased at 24 h after injury in either tissue homogenate or mitochondrial fractions. These results indicate that by 24 h post-injury ROS-induced protein oxidation is more prominent compared to lipid oxidation, indicating a critical temporal distinction in secondary pathophysiology that is critical in designing therapeutic approaches to mitigate consequences of oxidative stress.

Document Type

Article

Publication Date

8-2016

Notes/Citation Information

Published in Redox Biology, v. 8, p. 59-67.

© 2016 The Authors.

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Digital Object Identifier (DOI)

https://doi.org/10.1016/j.redox.2015.12.011

Funding Information

This study was supported by NIH/NINDS R01NS069633 (A.G.R. and P.G.S.), The Craig H. Neilsen Foundation 260771 (S.P.), The Craig H. Neilsen Foundation 190115 (A.G.R.) and NIH/NINDS P30 NS051220 (E.D.H).

Repository Citation

Visavadiya, Nishant P.; Patel, Samir P.; VanRooyen, Jenna L.; Sullivan, Patrick G.; and Rabchevsky, Alexander G., "Cellular and Subcellular Oxidative Stress Parameters Following Severe Spinal Cord Injury" (2016). Spinal Cord and Brain Injury Research Center Faculty Publications. 13.
https://uknowledge.uky.edu/scobirc_facpub/13