title: microRNA sequencing of human osteoblast in hypoxia 4 hours
design: Oxygen is a main participator of energy metabolism pathway energy in the human body. The degeneration of organ function may happen when Microcirculation doesn't get enough oxygen. And the consequence can be serious if there are no intervention for saving, sometimes even causing the patients  death.  Although hypoxia damage has been proved on many studies we cannot ignore the physiopathology value of its existence. The cells will be activated by hypoxia when a mass of bioactive factors are released. Some of these may mediate the tissue healing and control the derailed inflammation response. However, the benefit will be restricted by cells death in excessive hypoxia. Based on this, we did a pre-primary experiments to explore how to exert the biggest influence on hypoxia. Methods-The third generation of human osteoblasts were distributed into three groups. Cells of control group were normally cultured (high-sugar DMEM, 10% FBS and 1% penicillin streptomycin). Cells of hypoxia group were cultured in low-sugar DMEM (Invitrogen, USA) and 200 mol/L CoCl2 (sigma, USA) 4 hours and normally 4 hours. Cells of the Repeated brief hypoxia (RBH) group were under 3 times RBH (cultured in high-sugar DMEM, 10% FBS and 1% penicillin streptomycin 10min and then were washed by phosphate buffer saline 3 times. Then cultured in low-sugar DMEM and 200 mol/L CoCl2 10min and were washed with phosphate buffer saline 3 times. The above management was RBH 1 time), then under hypoxia management 4 hours and normally 4 hours. In the end, the related miRNA were detected by high-throughput sequencing. Results- High-throughput sequencing of microRNA shown the expression of 2763 miRNAs. Further, we identified that hypoxia 4 hours made 60 and 56 target genes up-regulated and down-regulated respectively compared with normally cultured cells (p<0.05). RBH 3 times + hypoxia 4 hours made 68 and 127 target genes up-regulated and down-regulated respectively compared with normally cultured cells (p<0.05). RBH 3 times + hypoxia 4 hours made 37 and 105 target genes up-regulated and down-regulated respectively compared with hypoxia 4 hours (p<0.05). Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analysis indicated relevant biological processes.Conclusion-Hypoxia 4 hours increased the cell death rate but promoted some bioactive factors secrete. High-throughput sequencing of microRNA shown that many miRNA participated in these effects to provide the basis for our future research.
identifiers.namespace: SUB3867916
identifiers.id: WANG 2