摘要
目的:研究CXC-型趋化因子4(CXCL4)对体外核因子κB受体活化因子配体(RANKL)诱导小鼠骨髓来源单核巨噬细胞(BMDM)向破骨细胞分化的影响。方法:无菌分离小鼠BMDM,使用50 ng/mL M-CSF以及不同浓度的CXCL4刺激细胞,CCK-8实验检测细胞增殖活性,Transwell迁移实验检测细胞的迁移能力。在M-CSF及RANKL均存在的前提下,以不同浓度的CXCL4作为干预因素,体外诱导细胞分化为破骨细胞,另设组加入BX471阻断剂处理细胞。TRAP染色法观察破骨细胞的形态并统计破骨细胞数目,RT-PCR检测JDP-2、NFATc1、c-Fos、MMP-9、TRAP、CTSK mRNA的表达情况。结果:不同浓度梯度的CXCL4均可增强BMDM的增殖活性及迁移能力;在RANKL及M-CSF均存在的前提下,加入不同浓度CXCL4的各实验组的TRAP染色阳性的细胞数量明显多于对照组(P<0.05),而在BX471阻断剂预处理组中TRAP染色阳性的细胞数量低于各实验组,但高于对照组(P<0.05)。此外,CXCL4能够上调破骨相关基因的表达,而BX471则能部分抑制上述mRNA表达的增高(P<0.05)。结论:CXCL4能提高BMDM的增殖及迁移能力,并促进RANKL介导的BMDM向破骨细胞分化,其促进作用可能与CXCL4/CCR1通路有关。
Objective:To investigate the effect of CXC-type chemokine 4(CXCL4)on the differentiation of mouse bone marrow-derived mononuclear macrophages(BMDM)into osteoclasts induced by Receptor Activator of Nuclear Factor-κB Ligand(RANKL)in vitro.Methods:Mouse BMDMs were isolated aseptically,then stimulated with 50ng/mL M-CSF and different concentrations of CXCL4.The proliferation activity of cell was examined by CCK-8 assay,and the migration ability was detected by Transwell migration assay.Under the the presence of both M-CSF and RANKL,different concentrations of CXCL4 were used as intervention factors to induce cell differentiation into osteoclasts in vitro,and another group was treated with BX471 blocker.The morphology of osteoclasts was observed by TRAP staining,the number of osteoclasts was counted.The mRNA expressions of JDP-2,NFATc1,c-Fos,MMP-9,TRAP and CTSK were detected by RT-PCR.Results:Different concentrations of CXCL4 could enhance the proliferative activity and the migration ability of BMDM.Under the presence of both RANKL and M-CSF,the number of TRAP-positive cells in the experimental groups with different concentrations of CXCL4 was significantly higher than that in the control group(P<0.05).The number of TRAP-positive cells in the BX471 blocker pretreatment group was lower than that in the experimental groups,but higher than that in the control group(P<0.05).In addition,CXCL4 up-regulated the expression of osteoclast-related genes,while BX471 partially inhibited the increased expression of these mRNA(P<0.05).Conclusion:CXCL4 can enhance the proliferation and migration abilities of BMDM and promote RANKL-mediated differentiation of BMDM into osteoclasts.The promotion may be related to CXCL4/CCR1 pathway.
作者
袁历峰
王佐林
YUAN Li-feng;WANG Zuo-lin(Department of Implant,School and Hospital of Stomatology,Tongji University,Shanghai EngineeringRearch Center of Tooth Restoration and Regeneration,Shanghai 200072,China)
出处
《口腔颌面外科杂志》
CAS
2018年第6期301-308,共8页
Journal of Oral and Maxillofacial Surgery
基金
中央高校基本科研业务费专项资金(1504219037)
