Interference Down-regulation Of Bone Remodeling Factors Associated With M-CSF And TNFSF11

Objective: To analyse the dynamic relationship between macrophage colony-sti mulating factor(M-CSF),RANKL mRNA and other related osteogenesis and osteo clast genes after down-regulation in order to explore the role of M-CSF and RAN KL in bone remodeling.Methods: The down-regulated M-CSF,RANKL shRNA a nd negative control recombinant plasmids were constructed,experimental groups wh ich is M-CSF shrna and negative control recombinant plasmid group,RANKL shrn a and negative control recombinant plasmid group.The recombinant plasmids were transfected into bone marrow stem cells by LIP3000.RNA was extracted 48 hour s later and reverse transcription was performed.The expression of osteogenic genes RUNX2,OSX,ALP,OCN,COL1,BMP2,and the expression of osteoclast genes RANKL,IL-1,IL-6,IL-24,TNF? and M-CSF mRNA were detected by RT-PCR.Results: M-CSF?RANKL shRNA compared with the negative control group,the expression of RANKL mRNA in the transfection group decreased(P < 0.05),IL-1,TNF?,M-CSF were down-regulated,while IL-6,IL-24 remained unchanged.Com pared with the negative control group,the expression of M-CSF mRNA in the tran sfected group was decreased,and the expression of OSX,OCN was up-regulated(P < 0.05),BMP2,RUNX2,ALP,COL1,basically unchanged.Conclusions: RAN KL is associated with IL-1,TNF? and M-CSF and is related to osteoclasts.The re lationship with IL-6,IL-24 is not clear.M-CSF is associated with RANKL and is related to osteoclasts.There are correlations between OSX,OCN in osteogenic path ways.The relationship with RUNX2,ALP,COL1,BMP2,is not clear.