Study On The Molecular Mechanism Of Hypoxia Promoting Osteoclast Formation

Objective: To investigate the effect of hypoxia on hypoxia mouse monocytes RAW264.7 in inducing factor-la(hypoxia inducible factor 1aloha,HIF,LA),transcription factor binding protein J(Recombinatation composite signal binding,protein-j,RBP-J,T)activated nuclear factor 1(nuclear factor activated T cell C1,NFATC1)effect and mi RNAs expression.To further explore the mechanism of hypoxia induced osteoclast differentiation.Methods: This experiment in vitro hypoxia environment(3% oxygen)and normal oxygen environment(20% oxygen)RAW264.7 mononuclear cells cultured under the mouse,and receptor activator of nuclear factor kappa B ligand(receptoractivator of B ligand RANKL NF-K,)induced at each time point(0h,3h,6h24 h,12h,and 48H)were collected,respectively.The following experiments: 1 by quantitative real-time PCR to detect the hypoxia inducible factor-la(HIF-LA),the expression of transcription factor RBP-J,NFATC1;2 by gene chip screening mi RNAs related to hypoxemia,again expression of mi RNAs gene by real-time quantitative PCR assay method for screening the.Results: 1.In vitro hypoxia environment(3% oxygen)and normal oxygen environment(20% oxygen)RAW264.7 mononuclear cells cultured under the mouse,and receptor activator of nuclear factor kappa B ligand(RANKL)induced by,at each time point(0h,3h,6h,12 h,24h,48h)collection cells using fluorescence PCR(Q-PCR)to detect the expression of HIF-l RAW264.7 in mononuclear cells in RBP-J,NFATC1,alpha m RNA,RAW264.7 mouse monocytes HIF-1a expression in hypoxia and normoxia was gradient increases,mouse monocyte RAW264.7 in normoxia and hypoxia in cultured 48 h,the expression of HIF-1a in hypoxia was significantly higher than that in normoxia,there was significant difference between two groups,(P < 0.05);mouse monocytes RAW264.7 in normoxia and hypoxia training 48 h,expression in hypoxia cells in RBP-J was significantly lower than that in normoxia,There was significant difference between two groups(P < 0.05);RAW264.7 mice mononuclear cells cultured in 12 h and 24 h in normoxia and hypoxia,expression in hypoxia cells in NFATC1 was significantly lower than that in normoxia,there was significant difference between two groups(P < 0.05).2.In vitro hypoxia environment(3% oxygen)and normal oxygen environment(20% oxygen)RAW264.7 mononuclear cells cultured under the mouse,and receptor activator of nuclear factor kappa B ligand(RANKL)induced by,at each time point(0h,3h,6h,12 h,24h,48h)were collected.The use of fluorescent PCR(Q-PCR)to detect the expression of RAW264.7 in mononuclear cells in mi R191,mi R127,mi R5099,m RNA,mouse monocyte RAW264.7 in normoxia and hypoxia in cultured48 h,the expression of mi R127 in hypoxic environment was significantly lower than that in normoxia,there was significant difference between two groups(P < 0.05);RAW264.7 mice mononuclear cells under hypoxia and normoxia environment(6h,12 h,24h)time point,the expression of mi R191 is gradient increases,while the RAW264.7 mice mononuclear cells in normoxia and hypoxia training 48 h,expression of mi R191 in normal oxygen environment Increased expression of hypoxia decreased,normoxia was significantly higher than that in hypoxia,there was significant difference between two groups,(P < 0.05);mouse monocytes RAW264.7 in normoxia and hypoxia in cultured 48 h,the expression of mi R5099 in hypoxic environment was significantly lower than that in normoxia,there was significant difference between two groups(P.< 0.05).Conclusion: 1.in vitro hypoxia and normoxia cultured mononuclear cell RAW264.7 at different time periods,cells in hypoxia inducible factor-l alpha(HIF-l alpha),there are differences in the expression of the transcription factor RBP-J and NFATc1,further explained that hypoxia stimulates osteoclast formation.2.in vitro hypoxia and normoxia cultured mononuclear cell RAW264.7 at different time periods,cells in mi RNAs(mi R191,mi R127,mi R5099)expression differences,further hypoxia can promote osteoclast formation by affecting the expression of mi RNAs.