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

摘要：

【摘要】 目的 观察高糖及TNF-α的培养条件对RAW264.7细胞向破骨细胞诱导分化的影响。方法 在正常、高糖(30 mmol/L)及TNF-α(10 μmol/L)条件下培养RAW264.7细胞后,加入浓度为100 ng/mL的细胞核转录因子κB受体激活物的配体(receptor activator of NF-κB ligand, RANKL)为诱导剂,诱导RAW264.7向破骨细胞分化;诱导9天后,抗酒石酸酸性磷酸酶(TRAP)染色,比较各组TRAP+细胞数,RT-PCR及Western Blot检测各组破骨细胞标志基因CTR和MMP-9的表达。结果 不同的培养条件下RANKL均能诱导RAW264.7分化为成熟的破骨细胞,其中TNF-α环境中RAW264.7形成的TRAP+阳性细胞数、CTR和MMP-9的表达最高,而在高糖环境下最低。结论 TNF-α可以促进RAW264.7向破骨细胞分化,而高糖对这个过程可能是抑制作用,这一现象符合Ⅰ型和Ⅱ糖尿病患者骨质破坏的表现;高糖及TNF-α的培养条件下RANKL对RAW264.7的作用可模拟糖尿病足病变微环境中OC的诱导分化的过程。

关键词: 高糖, TNF-α, RAW264.7, 破骨细胞, 糖尿病足