CD147对体外破骨细胞分化成熟的影响  

Effect of CD147 on differentiation and maturation of osteoclasts in vitro

作　　者：廖通权[1] 刘玉刚[1] 胡旭[1] 张莹[1] 余世明[1] 初同伟[1] 周跃[1] 

机构地区：[1]第三军医大学新桥医院骨科,重庆400037

出　　处：《第三军医大学学报》2010年第24期2623-2626,共4页Journal of Third Military Medical University

摘　　要：目的建立破骨细胞(osteoclasts,OCs)体外分化成熟的模型,研究CD147对OCs分化成熟的影响。方法从外周血分离单个核细胞贴壁培养,使用破骨细胞分化因子(receptor activator of nuclear factor-κβligand,RANKL)和巨噬细胞集落刺激因子(macrophage colony stimulating factor,M-CSF)诱导OCs生成。实验分为对照组、诱导组(RANKL+M-CSF)及阻断组(RANKL+M-CSF+CD147Ab),应用免疫荧光染色和流式细胞术检测CD147在破骨前体细胞(osteo-clast precursor cells,OPCs)的表达;抗酒石酸酸性磷酸酶(tartrate-resistant acid phosphatase,TRAP)染色观察OCs的分化成熟情况及Real-time PCR技术检测CD147及TRAP mRNA在OPCs中的表达变化。结果①CD147分子在OPCs细胞膜上表达。②流式细胞术测得细胞膜表面平均荧光强度对照组(159.55±1.65)、诱导24 h组(171.18±3.46)、诱导48 h组(178.47±4.59)逐渐增强(P<0.05),而阻断24 h组(117.69±9.01)显著减弱(P<0.01)。③阻断组TRAP染色阳性细胞(细胞核≥3)数显著少于诱导组(P<0.01),且OCs体积小。④在24、48 h诱导组CD147 mRNA相对表达量[(1.393±0.248)、(1.772+0.326)]及TRAP mRNA的相对表达量[(1.810±0.398)、(2.608±0.906)]在OPCs诱导分化过程逐渐增高,均高于相应时相点阻断组(P<0.05);且TRAP mRNA相对表达量与CD147 mRNA相对表达量呈正相关(r=0.833,P<0.01)。结论 CD147分子在OPCs细胞膜上表达且参与OCs分化成熟;抑制CD147的表达,OCs出现成熟障碍。Objective To establish a model of differentiation and maturation of osteoclasts（OCs） in vitro and to explore the effect of CD147 on the differentiation and maturation of OCs.Methods Peripheral blood mononuclear cells（PBMCs） were isolated from the fresh blood samples of 10 healthy volunteers,and then OCs were induced by using medium with macrophage colony stimulating factor（M-CSF） and receptor activator of nuclear factor-kappaΒ ligand（RANKL）.PBMCs were divided into 3 groups,control group,induction group（RANKL＋M-CSF） and blocking group（RANKL＋M-CSF＋CD147Ab）.Expression of CD147 was determined by immunofluorescence staining and flow cytometry,and mRNA levels of CD147 and tartrate-resistant acid phosphatase（TRAP） were detected by real-time PCR.The differentiation and maturation of OCs were observed by TRAP staining.Results CD147 was expressed on the membrane of osteoclast precursor cells（OPCs） in all groups.The average fluorescence intensity（AFI） of OPCs surface was gradually increased in control group（159.55±1.65）,and in induction group in 24 h（171.18±3.46） and 48 h（178.47±4.59）（P0.05）,but was significantly reduced in blocking group in 24 h（117.69±9.01,P0.01）.The TRAP positive cells（nuclei≥3） in the blocking group was significantly lesser and with smaller cell body compared with those in the induction group（P0.01）.The mRNA levels of CD147 [（1.393±0.248）,（1.772±0.326）] and TRAP [（1.810±0.398）,（2.608±0.906）] of induction group in 24 and 48 h after the induction were gradually increased in OPCs and were higher than those in the blocking group at corresponding time points（P0.05）.Moreover,the mRNA levels of CD147 was significantly positively correlated to the mRNA levels of CD147（r=0.833,P0.01）.Conclusion CD147 is expressed on the membrane of OPCs and involved in the differentiation and maturation of OCs.Inhibiting CD147 expression may result in dysmaturity of OCs.

关 键 词：破骨细胞 CD147 分化成熟 

分 类 号：R322.71[医药卫生—人体解剖和组织胚胎学] R329.21[医药卫生—基础医学]