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Chem Biol Interact. 2015 Jul 5;236:57-66. doi: 10.1016/j.cbi.2015.04.010. Epub 2015 Apr 30.

Proteomic analysis of the mouse brain after repetitive exposure to hypoxia.

Author information

1
Department of Pharmacology, School of Basic Medical Sciences, Capital Medical University, Beijing 10069, China.
2
Department of Pharmacology, School of Basic Medical Sciences, Capital Medical University, Beijing 10069, China; Beijing Laboratory for Biomedical Detection Technology and Instrument, Beijing 100069, China.
3
Department of Pharmacology, School of Basic Medical Sciences, Capital Medical University, Beijing 10069, China; Beijing Laboratory for Biomedical Detection Technology and Instrument, Beijing 100069, China. Electronic address: xuem@ccmu.edu.cn.

Abstract

Hypoxic preconditioning (HPC) is known to have a protective effect against hypoxic damage; however, the precise mechanisms involved remain unknown. In this study, an acute and repetitive hypoxia mouse model, two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) coupled with matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/TOF-MS), and Western blot experiments were used to identify the differential expression of key proteins in the mouse brain during HPC. Approximately 2100 2D-DIGE spots were observed following gel imaging and spot detection. Significant differences (p < 0.05) in the expression of 66 proteins were observed between the 3× HPC treatment group and the control group, 45 proteins were observed between the 6× HPC treatment group and the control group, and 70 proteins were observed between the 3× HPC treatment group and the 6× HPC group. Consistent results among Western blot, 2D-DIGE and MS methods were observed for the proteins, ATP synthase subunit alpha, malate dehydrogenase, guanine nucleotide-binding protein subunit beta-1 and proteasome subunit alpha type-2. The proteins associated with ATP synthesis and the citric acid cycle were down-regulated, while those linked to glycolysis and oxygen-binding were up-regulated. This proteomic analysis of the mouse brain after HPC furthers understanding of the molecular pathways involved in the protective effect of HPC and these findings provide new insight into the mechanisms of hypoxia and HPC.

KEYWORDS:

2D-DIGE; Hypoxic preconditioning; MALDI-TOF/TOF-MS; Mouse brain; Proteomics; Western blot

PMID:
25937538
DOI:
10.1016/j.cbi.2015.04.010
[Indexed for MEDLINE]

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