Abstract
The purpose of this study was to identify the key genes and pathways involved in hypoxic preconditioning of osteoblasts. Cells of the hypoxic-preconditioning group underwent 10 min of hypoxic cultivation/10 min normoxic cultivation three times before 4 h of hypoxic cultivation. The long-time hypoxia group underwent 4 h of hypoxic cultivation, and the control group cells underwent 4 h of normoxic cultivation. Differences in miRNAs expressed were analyzed by high-throughput sequencing. MiRwalk 3.0 was used to predict the target genes and conduct a Gene Set Enrichment Analysis. The STRING database and Cytoscape were used to explore the hub genes. A total of 8635 genes and 121 pathways are enriched in the hypoxic-preconditioning compared with the long-time hypoxia group, including the hub genes Utp6, Eif3a, Etf1, Rrp12 and Mkks. 8,476 target genes and 121 pathways are enriched in the hypoxic-preconditioning group compared with the control group, including the hub genes Chd7, Cebpa and Fosb, and 9,315 target genes and 131 pathways are enriched in the long-term hypoxia group compared with the control group, including the hub genes Dcun1d2, Ube2d1 and Frk. Hypoxia preconditioning improved cell viability (p < 0.01), enhanced ALP, IL10 and VEGF expression, and inhibited IL6 expression (p < 0.05). Hypoxic preconditioning is an effective method for promoting osteoblast activity.
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