不同浓度地西他滨对破骨细胞分化的影响  

Effect of different concentrations of decitabine on osteoclast differentiation

作　　者：糜宝国[1] 关邯峰[1] 刘常宇[1] 雷琢玮[1] 宋超[1] 刘慧勇[1] 邓杝 熊伟[1] 廖晖[1] 方忠[1] 李锋[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院骨科,武汉430030

出　　处：《骨科》2016年第2期120-124,共5页ORTHOPAEDICS

基　　金：中央高校基本科研业务费专项基金资助(2014TS074);湖北省自然科学基金(2014CFB196)

摘　　要：目的探讨不同浓度梯度地西他滨对破骨细胞形成、活性及吸收功能的影响。方法不同浓度地西他滨(0、0.1、0.25和0.5μmol/L)处理单核巨噬(RAW264.7)细胞。通过4’,6-联眯-2-苯基吲哚(4,6-Diamidino-2-phenylindole dihydrochloride,DAPI)染色和微丝绿色荧光探针(F-actin-Trakcer Green)染色后观察F-actin环的形成即破骨细胞轮廓;抗酒石酸酸性磷酸酶检测试剂盒检测细胞上清中的抗酒石酸酸性磷酸酶活性,骨板吸收实验检测破骨细胞的骨吸收能力;Q-PCR实验检测破骨细胞标志基因抗酒石酸酸性磷酸酶(tartrate-resistant acid phosphatase,TRAP)、组织蛋白酶K(cathepsin K,CK)和脊髓基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)的m RNA表达。结果不同浓度地西他滨抑制核因子NF-κB配体激活因子(receptor activator of NF-κB ligand,RANKL)诱导RAW264.7细胞形成F-actin环,降低了破骨细胞的TRAP酶活性,抑制了破骨细胞的骨吸收能力,同时也下调了破骨细胞标志基因TRAP、CK和MMP-9的m RNA表达。且随着药物浓度的增高,上述的抑制作用越明显。结论地西他滨抑制破骨细胞形成、活性和骨吸收能力,且这种抑制作用随着药物浓度的增加而逐渐增强。Objective To explore the effect of different concentrations of decitabine on osteoclast for-mation, activity and osteoclasts resorptive capacity. Methods RAW264.7 cells were treated with decitabine atdifferent concentrations（0, 0.1, 0.25, 0.5 μmol/L）. Osteoclast differentiation was assessed by DAPI and F-actin-Trakcer Green staining tests. Tartrate-resistant acid phosphatase kit was used to evaluate TRAP activity. Osteo-clasts resorptive capacity was detected by bone resorption test. The expression of osteoclast specific genes likeTRAP, capthesin K, and MMP- 9 m RNA was detected by RT- PCR. Results Different concentrations ofdecitabine inhabited RANKL-stimulated osteoclastogenesis in RAW264.7 cell culture, as manifested by de-crease of F-actin rings formation, TRAP activity and osteoclasts resorptive capacity in a dose-dependent man-ner, and also down-regulated TRAP, CK, and MMP-9 m RNA expression. Conclusion Decitabine inhabited os-teoclast formation, activity and osteoclasts resorptive capacity in a dose-dependent manner.

关 键 词：破骨细胞 细胞分化 RNA干扰 

分 类 号：R68[医药卫生—骨科学]