抄録
Hypoxia preconditioning (HPC) is known to have protective effect against hypoxia damage. Up to date, the exact molecular mechanism of HPC still remains unclear. In this study, the acute and repetitive hypoxia and HPC models using mice, zebrafish and PC12 cell were successfully constructed. Microarray and multiple bioinformatics were used to identify the differentially expressed genes, pathways and biological processes related to HPC. 2D-DIGE coupled with MALDI-TOF/TOF-MS and Western blot experiments were used to identify the differential expression of key proteins. The UPLC-HRMS based metabolomics was utilized to explore the key endogenous metabolites and pathways related to HPC. The results indicated that at least fourteen of these genes were the related genes for HPC, including Cacna2d1, Grin2a, Npy1r, Mef2c, Epha4, Rxfp1, Chrm3, Pde1a, Atp2b4, Glra1, Idi1, Fgf1, Grin2b and Cda. The Plcb1 gene is a hub gene and a key node in these signal-networks. The functions of these differentially expressed genes mainly involved the MAPK pathway, ion transport, neurotransmitter transport and neuropeptide signal pathway. 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. The levels of phenylalanine, valine, proline, leucine and glutamine were up-regulated, while that of creatine was decreased. The aminobutyric acid was markedly decreased in HPC brain. The sphingolipid metabolic pathways were also noticed under HPC, suggesting that C24:1-Cers played a critical role in HPC endogenous protective mechanism. Our data provided an important insight to further reveal the protection mechanism of HPC.
