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

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...

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

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(P<0.05),and the mRNA expression levels of MMP-2 and MMP-9 were positively correlated with those of CD147(r=0.818,P<0.01;r=0.758,P<0.01) 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(P<0.05).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,P>0.0.05),but the protein expression levels of MMP-9 at 48 h were higher than that at 24 h(t=7.589,P<0.05).Conclusion Exogenous CD147 protein can promote the synthesis and secretion of MMP-9 and MMP-2 during the differentiation and maturation of OCs.However,the secretion increase of MMP-2 is not time-dependent.