高糖及TNF-α对RAW264.7细胞向破骨细胞分化的影响  

Effect of high glucose and TNF-α on the differentiation of RAW264.7 cells to osteoclast

作　　者：侯念宗 刘文宙 陈炳豪 陈皓[1] Aditya 宋卫东[1] 

机构地区：[1]广州中山大学孙逸纪念医院骨外科,510120

出　　处：《岭南现代临床外科》2013年第3期172-177,共6页Lingnan Modern Clinics in Surgery

基　　金：广东省科技计划社会发展项目(2011B031800217)

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(30mmol/L)及TNF-α(10μmol/L)条件下培养RAW264.7细胞后,加入浓度为100ng/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。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 type Ⅰ and Ⅱ 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.

关 键 词：高糖 TNF-Α RAW264 7 破骨细胞 糖尿病足 

分 类 号：R681[医药卫生—骨科学]