关键词: 高糖, TNF-α, RAW264.7, 破骨细胞, 糖尿病足

Abstract:

【Abstract】 Objective To observe the effect of high glucose and TNF-α on the differentiation process of RAW264.7 cells to osteoclasts. Methods RAW264.7 cell were cultured in  DMEM medium  under the  condition of normal,.high glucose(30 mmol/L) and TNF-α(10 μmol/L) respectively and RANKL(receptor activator of NF-κB ligand,.100 ng/mL).was added and used to induce the the differentiation process of RAW264.7 to osteoclast..After 9 days,.the TRAP positive cells after tartrate resistant acid phosphatase (TRAP) staining and the expression of CTR and MMP-9 detected by RT-PCR and Western Blot were compared among the three culture conditions of normal,.high glucose and TNF-α respectively. Results RANKL could induce the differentiation of RAW264.7 to osteoclasts under different culture conditions and the TRAP positive cells and the expression of CTR and MMP-9 were the most in the environment of TNF-α,.but the least of high glucose. Conclusions TNF-α can promote differentiation of RAW264.7 to osteoclast,.but high glucose is inhibitory,.which can explain bone damage of patients with typeⅠandⅡ diabetes. The induction of differentiation of RAW264.7 to osteoclast by RNAKL under different culture conditions of normal,.high glucose and TNF-α can simulate the process of differentiation of precursor cells to osteoclasts in the patients with diabetic foot.

Key words: High glucose, TNF-α, RAW264.7, Osteoclast, Diabetic foot