摘要
目的探讨阳离子通道阻断剂钌红对破骨细胞分化的影响及其机制。方法钌红处理Raw246．7后，50μg／L核因子．KB受体活化因子配体（RANKL）诱导6d，抗酒石酸酸性磷酸酶（TRAP）染色观察TRAP＋（核93）细胞的形成；TRAP试剂盒检测细胞中TRAP酶的活性；实时定量逆转录聚合酶链反应（FIT—qPCR）检测破骨细胞特异性标志基因TRAP、组织蛋白酶K（CK）和金属基质蛋白酶-9（MMPO）mRNA表达量；RT．qPCR、Westernblot分别检测核因子-κB受体活化因子（RANK）在mRNA和蛋白水平上的表达。结果钌红促进RANKL诱导Raw246．7细胞形成TRAP阳性多核细胞，对照组TRAP吸光度（A）值为0．33±0．04，而钌红组TRAPA值为0．43±0．03；同时也上调了破骨细胞标志性基因TRAP、CK和MMPO的mRNA表达以及RANK在mRNA和蛋白水平的表达。各指标在两组中的差异有统计学意义（P〈0105）。结论钌红促进Raw246．7向破骨细胞分化，这一过程与钌红上调RANK表达有关，即与RANK—RANKL轴相关。
Objective To investigate the effects of ruthenium red on the differentiation of osteo- clasts and relative mechanism. Methods Receptor activator of nuclear factor-KB ligand （RANKL） was used to stimulate osteoclast differentiation. Osteoclast differentiation was assessed by tartrate-resistant acid phosphatase （TRAP） staining and the TRAP-positive cells were counted after exposure to 50 μg/L RANKL for 6 days. Tartrate-resistant acid phosphatase kit was used to evaluate TRAP activity. The expression of osteoclast specific genes [ TRAP, Capthesin K （ CK）, matrix metalloproteinase （MMP） -9 ] mRNA expres- sion was detected by using real-time quantitative reverse transcription-polymerase chain reaction （RT-qPCR）. The receptor activator of nuclear factor Kb（RANK） mRNA and protein expression was also detected. Results Ruthenium red promoted RANKL-stimulated osteoclast differentiation in Raw246.7 cell culture, as manifested by the increase of TRAP positive muhinucleated cells and TRAP activity. In control group the TRAP A was 0. 33 ± 0. 04, and that in ruthenium red group was 0. 43± 0. 03. Ruthenium also up-regulated TRAP, CK, MMP-9 and RANK mRNA expression, and increased RANK protein expression. The difference between two groups was significant （ P 〈 0. 05 ）. Conclusion Ruthenium red promoted oste- oclast differentiation, which might by related to RANK-RANKL axis.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2013年第4期784-786,共3页
Chinese Journal of Experimental Surgery
关键词
钌红
破骨细胞
分化
Ruthenium red
Osteoclastst
Differentiation
