不同浓度钛合金微粒对成骨细胞信号通路中转录表达因子RUNX2影响  

Different concentrations of titanium alloy particles on RUNX2 transcriptional expression of osteoblast signaling pathway

作　　者：孙国静[1] 杨书丰 郭亭[1] 赵建宁[1] 

机构地区：[1]第二军医大学南京临床医学院(南京军区南京总医院)骨科,南京210002 [2]解放军81医院骨科,南京210002

出　　处：《医学研究生学报》2013年第3期251-254,共4页Journal of Medical Postgraduates

基　　金：南京军区医学科技创新项目(11Z028);南京市科技发展项目(2012sc311020)

摘　　要：目的关节假体松动后出现骨溶解,如何有效阻止骨溶解,一直是个难题。文中通过探讨不同浓度钛合金微粒对转录因子RUNX2影响,进一步了解磨损微粒抑制成骨细胞作用机制。方法实验分为空白对照组和钛合金微粒处理组5μg/ml(A组)、10μg/ml(B组)、15μg/ml(C组)。采用细胞计数试剂盒-8法检测各组培养120 h成骨细胞增殖活性。分别采用RT-PCR和蛋白免疫印迹法(Western)检测培养120 h后成骨细胞的RUNX2 mRNA和蛋白表达。结果培养120 h后,与空白对照组相比,各钛合金处理组细胞增殖活性显著下降(P<0.05),B组、C组细胞中RUNX2 mRNA水平亦明显下降(P<0.01),并且RUNX2蛋白表达水平变化趋势与其mRNA的表达变化趋势一致。结论钛合金微粒降低成骨细胞增殖活性,下调成骨细胞RUNX2-转录因子的表达。钛合金微粒对RUNX2表达水平的调控可能是其抑制成骨细胞增殖的机制之一。Objective It is difficult to prevent osteolysis around joint prosthesis. The study aimed to investigate the effect of Titanium alloy particles （TAPs） at different concentrations on RUNX2 expression in MC3T3-E1 cells, and tried to explore the mecha- nism responsible for the decrease in osteoblast proliferation induced by TAPs. Methods Osteoblasts were cultured in medium con- taining different concentrations of TAPs. Experiments were cultured in medium without TAPs, while osteoblasts in groups A, B and C were cultured in medium containing TAPs at 5 μg/ml, 10 μg/ml and 15μg/ml, respectively. Cell Counting Kit 8 test was used to assay the cell proliferation after 120 h-incubation with TAPs. Furthermore, RUNX2 mRNA levels and its protein expression were detected by reverse transcription-polymerase chain reaction （RT-PCR） and Western blotting, respectively, at the time point of 120 h. Results Compared with the control ceils, after 120 h-incubation with TAPs, the proliferation of osteoblasts in group A, B or C was significantly decreased with comparison of the control ceils in control group （P 〈 0.05）. Meanwhile the RUNX2 mRNA levels in groups B and Cwere remarkably decreased by 120 h-exposure to TAPs （P 〈 0.01 ）. The RUNX2 protein levels were also down-regulated by TAPs, which was consistent with the changes in its mRNA levels. Conclusion The proliferation of osteoblasts was decreased by the exposure to TAPs in MC3T3-E1 cells. Further investigation revealed that RUNX2 mR- NA and protein levels were significantly down-regulated by TAPs.These data suggest that the inhibiting effects of TAPs on osteoblast proliferation were probably due to the suppression of RUNX2 expres- sion induced by TAPs in MC3T3-E1 cells.

关 键 词：钛合金颗粒 成骨细胞 RUNX2 

分 类 号：Q25[生物学—细胞生物学]