Proteomic analysis of the mouse brain after repetitive exposure to hypoxia.
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.
Department of Pharmacology, School of Basic Medical Sciences, Capital Medical University, Beijing 10069, China.
Can
C
Cui
Department of Pharmacology, School of Basic Medical Sciences, Capital Medical University, Beijing 10069, China.
Tao
T
Zhou
Department of Pharmacology, School of Basic Medical Sciences, Capital Medical University, Beijing 10069, China.
Jingyi
J
Li
Department of Pharmacology, School of Basic Medical Sciences, Capital Medical University, Beijing 10069, China.
Hong
H
Wang
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.
Xiaorong
X
Li
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.
Jie
J
Xiong
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.
Pingxiang
P
Xu
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.
Ming
M
Xue
D000222:Adaptation, Physiological; D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism* / Q000502:physiology; D004195:Disease Models, Animal; D015180:Electrophoresis, Gel, Two-Dimensional; D044387:GTP-Binding Protein beta Subunits; D020962:Heterotrimeric GTP-Binding Proteins / Q000378:metabolism; D002534:Hypoxia, Brain / Q000378:metabolism* / Q000503:physiopathology; D019194:Ischemic Preconditioning; D008297:Male; D008813:Mice, Inbred ICR; D024101:Mitochondrial Proteins / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D011506:Proteins / Q000032:analysis / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D012016:Reference Values; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D044387:GTP-Binding Protein beta Subunits; C491082:Gnb1 protein, mouse; D024101:Mitochondrial Proteins; D011506:Proteins; C000600515:PSA2 protein, mouse; D046988:Proteasome Endopeptidase Complex; D020962:Heterotrimeric GTP-Binding Proteins
2D-DIGE; Hypoxic preconditioning; MALDI-TOF/TOF-MS; Mouse brain; Proteomics; Western blot