Hypoxia preconditioning （HPC） is associated with many complicated pathophysiological and biochemical processes that integrated and regulated via molecular levels. HPC could protect cells, tissues, organs and systems...Hypoxia preconditioning （HPC） is associated with many complicated pathophysiological and biochemical processes that integrated and regulated via molecular levels. HPC could protect cells, tissues, organs and systems from hypoxia injury, but up to date, the molecular mechanism still remained unclear. The acute and repetitive hy- poxia preconditioning model was constructed and the related parameters were observed. The high-throughput mi- croarray analysis and multiple bioinformatics were used to explore the differentially expressed genes in HPC mice brain and the related gene network, pathways and biological processes related to HPC. The 2D-DIGE coupled with MALDI-TOF/TOF-MS was performed to identify these proteins that were differentially expressed during HPC. The UPLC-HRMS based metabolomics method was utilized to explore the key endogenous metabolites and metabolic pathways related to HPC. The results showed that （1） 1175 differentially expressed genes in HPC mice brain were identified. Fourteen of these genes were the related hub genes for HPC, including Cacna2dl, Grin2a, Npylr, Mef2c, Epha4, Rxfpl, Chrm3, Pdela, Atp2b4, Glral, Idil , Fgfl, Grin2b and Cda. The change trends of all the detected genes by RT-PCR were consistent with the data of gene chips. There were 113 significant functions up- regulated and 138 significant functions down-regulated in HPC mice. （2） About 2100 proteins were revealed via the gel imaging and spot detection. 66, 45 and 70 of proteins were found to have significantly difference between the control group and three times of HPC group, the control and six times of HPC, and the three times of HPC and six times of HPC group. （3）Some endogenous metabolites such as phenylalanine, valine, proline, leucine and glu- tamine were increased, while ereatine was decreased, both in HPC brain and heart; in addition, y-aminobutyric acid was markedly decreased in brain. The sphingolipid metabolic pathways were noticed due to the low p-value and high pathway impact. Especially, the sphingolipid compound sphingomyelin, ceramide, glucosyleeramide, galactosylceramide and laetosylceramide were mapping in this metabolic pathway. Interestingly, these sphingolipid metabolites with olefinic bond in the long fatty chain were up-regulated, while those sphingolipids without olefinic bond were down-regulated. The functions of these differentially expressed genes mainly involved the cellular proces- ses including MAPK pathway, ion transport, neurotransmitter transport and neuropeptide signal pathway. The pro- tein levels related the ATP synthesis and citric acid cycle decreased while the proteins with the glycolysis and oxy- gen-binding increased. Glutathione, GNBP-1 and GPD1L were related to preventing hypoxic damage. The results indicated that C24：l-Cers played a critical role in HPC and had potential in endogenous protective mechanism. The combinations of the system omies data of the different molecules were sufficient to give a further understanding of the molecular pathways affected by HPC. Our data provided an important insight to reveal the protection mechanism of HPC.展开更多
OBJECTIVE Hypoxia is associated with many complicated pathophysiological and biochemical processes that integrated and regulated via the key gene,protein and endogenous metabolite levels.Up to date,the exact molecular...OBJECTIVE Hypoxia is associated with many complicated pathophysiological and biochemical processes that integrated and regulated via the key gene,protein and endogenous metabolite levels.Up to date,the exact molecular mechanism of hypoxia still remains unclear.In this work,we further explore the molecular mechanism of hypoxia and adaption to attenuate the damage in zebrafish model that have potential to resist hypoxic environment.METHODS The hypoxic zebrafish model was established in different concentration of oxygen with 3%,5%,10%,21%in water.The brain tissue was separated and the RNA-seq was used to identify the differentially expressed genes.The related endogenous metabolites profiles were obtained by LC-HDMS,and the multivariate statistics was applied to discover the important metabolites candidates in hypoxic zebrafish.The candidates were searched in HMDB,KEGG and Lipid Maps databases.RESULTS The zebrafish hypoxic model was successfully constructed via the different concentration of oxygen,temperature and hypoxic time.The activities of the related hypoxic metabolic enzymes and factors including HIF-1a,actate dehydrogenase(LDH)and citrate synthase(CS)were evaluated.Significant differences(P<0.05 and fold change>2)in the expression of 422 genes were observed between the normal and 3% hypoxic model.Among them,201 genes increased depended on the lower concentration of oxygen.53 metabolites were identified that had significant difference between the hypoxia and control groups(P<0.05,fold change>1.5 and VIP>1.5).The ten key metabolites were increased gradually while six compounds were decreased.The endogenous hypoxic metabolites of phenylalanine,D-glucosamine-6P and several important lipids with the relevant hub genes had similar change in hypoxic model.In addition,the metabolic pathways of phenylalanine,glutamine and glycolipid were influenced in both the levels of genes and metabolites.CONCLUSION The up-regulation of phenylalanine,D-glucosamine-6P and lipid may have further understanding of protective effect in hypoxia.Our data provided an insight to further reveal the hypoxia and adaptation mechanism.展开更多
文摘Hypoxia preconditioning （HPC） is associated with many complicated pathophysiological and biochemical processes that integrated and regulated via molecular levels. HPC could protect cells, tissues, organs and systems from hypoxia injury, but up to date, the molecular mechanism still remained unclear. The acute and repetitive hy- poxia preconditioning model was constructed and the related parameters were observed. The high-throughput mi- croarray analysis and multiple bioinformatics were used to explore the differentially expressed genes in HPC mice brain and the related gene network, pathways and biological processes related to HPC. The 2D-DIGE coupled with MALDI-TOF/TOF-MS was performed to identify these proteins that were differentially expressed during HPC. The UPLC-HRMS based metabolomics method was utilized to explore the key endogenous metabolites and metabolic pathways related to HPC. The results showed that （1） 1175 differentially expressed genes in HPC mice brain were identified. Fourteen of these genes were the related hub genes for HPC, including Cacna2dl, Grin2a, Npylr, Mef2c, Epha4, Rxfpl, Chrm3, Pdela, Atp2b4, Glral, Idil , Fgfl, Grin2b and Cda. The change trends of all the detected genes by RT-PCR were consistent with the data of gene chips. There were 113 significant functions up- regulated and 138 significant functions down-regulated in HPC mice. （2） About 2100 proteins were revealed via the gel imaging and spot detection. 66, 45 and 70 of proteins were found to have significantly difference between the control group and three times of HPC group, the control and six times of HPC, and the three times of HPC and six times of HPC group. （3）Some endogenous metabolites such as phenylalanine, valine, proline, leucine and glu- tamine were increased, while ereatine was decreased, both in HPC brain and heart; in addition, y-aminobutyric acid was markedly decreased in brain. The sphingolipid metabolic pathways were noticed due to the low p-value and high pathway impact. Especially, the sphingolipid compound sphingomyelin, ceramide, glucosyleeramide, galactosylceramide and laetosylceramide were mapping in this metabolic pathway. Interestingly, these sphingolipid metabolites with olefinic bond in the long fatty chain were up-regulated, while those sphingolipids without olefinic bond were down-regulated. The functions of these differentially expressed genes mainly involved the cellular proces- ses including MAPK pathway, ion transport, neurotransmitter transport and neuropeptide signal pathway. The pro- tein levels related the ATP synthesis and citric acid cycle decreased while the proteins with the glycolysis and oxy- gen-binding increased. Glutathione, GNBP-1 and GPD1L were related to preventing hypoxic damage. The results indicated that C24：l-Cers played a critical role in HPC and had potential in endogenous protective mechanism. The combinations of the system omies data of the different molecules were sufficient to give a further understanding of the molecular pathways affected by HPC. Our data provided an important insight to reveal the protection mechanism of HPC.
基金supported by National Natural Science Foundation of China(81573683 and 81173121)
文摘OBJECTIVE Hypoxia is associated with many complicated pathophysiological and biochemical processes that integrated and regulated via the key gene,protein and endogenous metabolite levels.Up to date,the exact molecular mechanism of hypoxia still remains unclear.In this work,we further explore the molecular mechanism of hypoxia and adaption to attenuate the damage in zebrafish model that have potential to resist hypoxic environment.METHODS The hypoxic zebrafish model was established in different concentration of oxygen with 3%,5%,10%,21%in water.The brain tissue was separated and the RNA-seq was used to identify the differentially expressed genes.The related endogenous metabolites profiles were obtained by LC-HDMS,and the multivariate statistics was applied to discover the important metabolites candidates in hypoxic zebrafish.The candidates were searched in HMDB,KEGG and Lipid Maps databases.RESULTS The zebrafish hypoxic model was successfully constructed via the different concentration of oxygen,temperature and hypoxic time.The activities of the related hypoxic metabolic enzymes and factors including HIF-1a,actate dehydrogenase(LDH)and citrate synthase(CS)were evaluated.Significant differences(P<0.05 and fold change>2)in the expression of 422 genes were observed between the normal and 3% hypoxic model.Among them,201 genes increased depended on the lower concentration of oxygen.53 metabolites were identified that had significant difference between the hypoxia and control groups(P<0.05,fold change>1.5 and VIP>1.5).The ten key metabolites were increased gradually while six compounds were decreased.The endogenous hypoxic metabolites of phenylalanine,D-glucosamine-6P and several important lipids with the relevant hub genes had similar change in hypoxic model.In addition,the metabolic pathways of phenylalanine,glutamine and glycolipid were influenced in both the levels of genes and metabolites.CONCLUSION The up-regulation of phenylalanine,D-glucosamine-6P and lipid may have further understanding of protective effect in hypoxia.Our data provided an insight to further reveal the hypoxia and adaptation mechanism.
