CD147对体外破骨细胞分化过程中基质金属蛋白酶-2、基质金属蛋白酶-9分泌的影响  

Effect of exogenous CD147 protein on synthesis and secretion of matrix metalloproteinases-2 and 9 during differentiation and maturation of osteoclasts in vitro

作　　者：余世明[1] 初同伟[1] 廖通权[1] 胡旭[1] 刘玉刚[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》2011年第14期1492-1496,共5页Journal of Third Military Medical University

摘　　要：目的建立体外破骨细胞(osteoclasts,OCs)诱导分化模型,研究CD147对体外破骨细胞分化过程中基质金属蛋白酶-2(matrix metalloproteinases 2,MMP-2)和基质金属蛋白酶-9(matrix metalloproteinases 9,MMP-9)合成与分泌的影响。方法采用健康成人外周血分离单个核细胞贴壁培养,应用巨噬细胞集落刺激因子(macrophage colony stimulatingfactor,M-CSF)与NF-κB受体活化因子配体(receptor or activator of NF-κB ligand,RANKL)诱导单个核细胞向Ocs分化。实验分为对照组(RANKL+M-CSF)与蛋白诱导组(RANKL+M-CSF+CD147蛋白)。应用抗酒石酸酸性磷酸酶染色(tar-trate-resistant acid phosphatase,TRAP)及骨吸收实验检测鉴定OCs的分化成熟,Real-time PCR技术检测CD147、MMP-2和MMP-9 mRNA在破骨前体细胞(osteoclast precursor cells,OPCs)中表达变化,酶联免疫吸附实验(ELISA)检测细胞上清液中MMP-2、MMP-9的表达。结果①TRAP染色和骨吸收实验检测到破骨样细胞形成并具有骨吸收功能,建立体外Ocs诱导分化模型成功。②在24、48 h蛋白诱导组CD147 mRNA[(1.344±0.207)、(1.816±0.222)]、MMP-2 mRNA[(1.187±0.077)、(1.437±0.231)]及MMP-9 mRNA[(1.128±0.107)、(2.353±0.679)]的相对表达量均高于相应的对照组(P<0.05),且MMP-2、MMP-9 mRNA的相对表达量与CD147 mRNA的表达量呈正相关(r分别为0.818、0.758,P<0.01)。③24、48 h蛋白诱导组MMP-2蛋白[(386.71±40.66)、(367.35±20.72)μg/L]、MMP-9蛋白[(1 177.78±66.42)、(1 514.37±68.56)μg/L]表达均高于相应对照组(P<0.05),蛋白诱导组MMP-2蛋白的表达在24 h与48 h间无统计学差异(t=1.190,P>0.05),而MMP-9蛋白的表达在48 h显著高于24 h(t=7.589,P<0.05)。结论外源性CD147蛋白可以促进OCs分化成熟过程中MMP-2及MMP-9的合成与分泌,但MMP-2分泌的增加不呈时间依赖性。Objective To establish a model of differentiation and maturation of osteoclasts（OCs） in vitro,and to study the effect of CD147 on the synthesis and secretion of matrix metalloproteinases-2 and 9（MMP-2 and MMP-9） during the differentiation and maturation of OCs in vitro.Method Human peripheral blood mononuclear cells（PBMCs） were obtained from healthy adults by attachment culture.Macrophage colony stimulating factor（M-CSF） and receptor activator of nuclear factor-kappa Β ligand（RANKL） were used to induce the differentiation of PBMCs into OCs.PBMCs were divided into a control group（RANKL＋M-CSF） and a protein induction group（RANKL＋M-CSF＋CD147）.The differentiation and maturation of OCs were identified by tartrate-resistant acid phosphatase（TRAP） staining and bone resorption test.The mRNA expression levels of CD147,MMP-2 and MMP-9 in osteoclast precursor cells（OPCs） were detected by real-time PCR.The protein expression levels of MMP-2 and MMP-9 in the cell supernatant were detected by ELISA.Results OC-like cells were detected and showed a bone resorption function in TRAP staining and bone resorption test.The mRNA expression levels of CD147（1.344±0.207 and 1.816±0.222）,MMP-2（1.187±0.077 and 1.437±0.231） and MMP-9（1.128±0.107 and 2.353±0.679） in the protein induction group at 24 h and 48 h were higher than those in the control group（P0.05）,and the mRNA expression levels of MMP-2 and MMP-9 were positively correlated with those of CD147（r=0.818,P0.01;r=0.758,P0.01） The protein expression levels of MMP-2（386.71±40.66 μg/L and 367.35±20.72 μg/L） and MMP-9（1 177.78±66.42 μg/L and 1 514.37±68.56 μg/L） in the protein induction group at 24 h and 48 h were higher than those in the control group（P0.05）.The protein expression levels of MMP-2 in the protein induction group showed no significant difference at 24 h and 48 h（t=1.19,P0.0.05）,but the protein expression levels of MMP-9 at 48 h were higher than that at 24 h（t=7.589,P0.05）.Concl

关 键 词：CD147 破骨细胞 MMP-2 MMP-9 

分 类 号：R336[医药卫生—人体生理学] R329.28[医药卫生—基础医学]