title: Quantitative proteomics analysis to identify diffuse axonal injury biomarkers in rats using iTRAQ coupled LC-MS/MS.
abstract: Diffuse axonal injury (DAI) is fairly common during a traumatic brain injury (TBI) and is associated with high mortality. Making an early diagnosis, appropriate therapeutic decisions, and an accurate prognostic evaluation of patients with DAI still pose difficulties for clinicians. The detailed mechanisms of axonal injury after head trauma have yet to be clearly defined and no reliable biomarkers are available for early DAI diagnosis. Therefore, this study employed an established DAI animal model in conjunction with an isobaric tag for relative and absolute quantification (iTRAQ)-based protein identification/quantification approach. Alterations in rat cerebral protein expression were quantified using iTRAQ coupled LC-MS/MS, with differentially expressed proteins between the control groups, sham and sham-injured, and the injury groups, animals that died immediately post-injury and those sacrificed at 1h, 6h, 1d, 3d and 7d post-injury, identified. A total of 1858 proteins were identified and quantified and comparative analysis identified ten candidate proteins that warranted further examination. Of the ten candidate DAI biomarkers, four proteins, citrate synthase (CS), synaptosomal-associated protein 25 (Snap25), microtubule-associated protein 1B (MAP1B) and Rho-associated protein kinase 2 (Rock2), were validated by subsequent Western blot and immunohistochemistry analyses. Our studies not only identified several novel biomarkers that may provide insight into the pathophysiological mechanisms of DAI, but also demonstrated the feasibility of iTRAQ-based quantitative proteomic analysis in cerebral tissue research.
authors.affiliation: Department of Forensic Medicine, Faculty of Basic Medical Sciences, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Zhang
authors.affiliation: Faculty of Medical Technology, Chongqing Medical and Pharmaceutical College, Chongqing 401331, China.
authors.forename: Shisheng
authors.initials: S
authors.lastname: Zhu
authors.affiliation: Department of Forensic Medicine, Faculty of Basic Medical Sciences, Chongqing Medical University, Chongqing 400016, China; Chongqing Engineering Research Center for Criminal Investigation Technology, Chongqing 400016, China.
authors.forename: Yongguo
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Forensic Medicine, Faculty of Basic Medical Sciences, Chongqing Medical University, Chongqing 400016, China; Chongqing Engineering Research Center for Criminal Investigation Technology, Chongqing 400016, China.
authors.forename: Minzhu
authors.initials: M
authors.lastname: Zhao
authors.affiliation: Department of Forensic Medicine, Faculty of Basic Medical Sciences, Chongqing Medical University, Chongqing 400016, China; Chongqing Engineering Research Center for Criminal Investigation Technology, Chongqing 400016, China.
authors.forename: Meng
authors.initials: M
authors.lastname: Liu
authors.affiliation: Department of Forensic Medicine, Faculty of Basic Medical Sciences, Chongqing Medical University, Chongqing 400016, China; Chongqing Engineering Research Center for Criminal Investigation Technology, Chongqing 400016, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Gao
authors.affiliation: Chongqing Engineering Research Center for Criminal Investigation Technology, Chongqing 400016, China; Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Shijia
authors.initials: S
authors.lastname: Ding
authors.affiliation: Department of Forensic Medicine, Faculty of Basic Medical Sciences, Chongqing Medical University, Chongqing 400016, China. Electronic address: ljb2032008@163.com.
authors.forename: Jianbo
authors.initials: J
authors.lastname: Li
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002853:Chromatography, Liquid; D020833:Diffuse Axonal Injury / Q000378:metabolism*; D008297:Male; D013058:Mass Spectrometry; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D040901:Proteomics; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D015415:Biomarkers; D009419:Nerve Tissue Proteins
keywords: Biomarkers; Diffuse axonal injury; Proteomics; Rats; iTRAQ