Mechanism of participation of osteocytes in the formation of osteoclasts under hypoxia.

Jie Zhu; Yi Tang; Qing WU; Ying-chen JI; Fei-wu Kang

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Mechanism of participation of osteocytes in the formation of osteoclasts under hypoxia.

Author: Jie ZHU ¹ ; Yi TANG ¹ ; Qing WU ¹ ; Ying-Chen JI ¹ ; Fei-Wu KANG ¹
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1. Dept. of Oral and Maxillofacial Surgery, School and Hospital of Stomatology, Tongji University, Shanghai Engineering Research Center of Tooth Restoration and Regeneration, Shanghai 200072, China.
Publication Type:Journal Article
Keywords: deferoxamine mesylate; hypoxia-inducible factor-1α; receptor activator of nuclear factor-κB ligand
MeSH: Cell Differentiation; Cell Line; Humans; Hypoxia; Osteoclasts; Osteocytes
From: West China Journal of Stomatology 2019;37(5):463-468
CountryChina
Language:Chinese
Abstract: OBJECTIVE:To investigate the mechanism of the participation of osteocytes in the formation of osteoclasts under hypoxia.
METHODS:The 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.
RESULTS:DFO (100 μmol·L⁻¹) promoted the proliferation of MLO-Y4 at 24 h, which decreased with time (P<0.01). After the addition of soluble sRANKL, the formation of osteoclasts was significantly increased in the DFO group (P<0.001). The expression of RANKL mRNA in MLO-Y4 under 100 μmol·L⁻¹ DFO increased first and then decreased with the duration of hypoxia. This expression reached a peak at 24 h (P<0.01). Hypoxia up-regulated the expression of HIF-1α and RANKL protein (P<0.01). Under hypoxia, siHIF-1α downregulated the expression of HIF-1α and RANKL (P<0.01). siHIF-1α also decreased the number of osteoclasts (P<0.01).
CONCLUSIONS:Under hypoxia, MLO-Y4 could facilitate the formation of RANKL through upre-gulating the expression of HIF-1α protein, thereby accelerate the differentiation of RAW264.7 cells into osteoclasts.