低氧状态下骨细胞参与破骨细胞形成的作用机制研究

ObjectiveTo investigate the mechanism of the participation of osteocytes in the formation of osteoclasts under hypoxia.MethodsThe hypoxia culture system of osteocyte-like cell line MLO-Y4 was established by deferoxamine mesylate (DFO)in vitro. The proliferation of MLO-Y4 cells was examined by CCK-8 cell proliferation/toxicity assay. RAW264.7 cells were induced to osteoclasts by the conditioned medium containing the cultured MLO-Y4. Tartrate-resistant acid phosphatase (TRAP) staining was performed on day 7. Quantitative real-time fluorescence polymerase chain reaction, immunofluorescence, and Western blot were used to detect the expression levels of hypoxia-inducible factor (HIF)-1α and receptor activator of nuclear factor-κB ligand (RANKL) in MLO-Y4 under hypoxia. The effects of siHIF-1α on the expression levels of HIF-1α and RANKL in MLO-Y4 under the same conditions were detected.ResultsDFO (100 μmol·L^-1) promoted the proliferation of MLO-Y4 at 24 h, which decreased with time (PP-1DFO increased first and then decreased with the duration of hypoxia. This expression reached a peak at 24 h (PPPPConclusionUnder hypoxia, MLO-Y4 could facilitate the formation of RANKL through upregulating the expression of HIF-1α protein, thereby accelerate the differentiation of RAW264.7 cells into osteoclasts.