[1]余世明,初同伟,廖通权,等.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[J].陆军军医大学学报(原第三军医大学学报),2011,33(14):1492-1496.
 Yu Shiming,Chu Tongwei,Liao Tongquan,et al.Effect of exogenous CD147 protein on synthesis and secretion of matrix metalloproteinases-2 and 9 during differentiation and maturation of osteoclasts in vitro[J].J Amry Med Univ (J Third Mil Med Univ),2011,33(14):1492-1496.
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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(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
33卷
期数:
2011年第14期
页码:
1492-1496
栏目:
论著
出版日期:
2011-07-30

文章信息/Info

Title:
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:
余世明初同伟廖通权胡旭刘玉刚周跃
第三军医大学新桥医院骨科
Author(s):
Yu Shiming Chu Tongwei Liao Tongquan Hu Xu Liu Yugang Zhou Yue
Department of Orthopedics, Xinqiao Hospital, Third Military Medical University,Chongqing, 400037, China
关键词:
CD147破骨细胞MMP-2MMP-9
Keywords:
CD147 osteoclasts MMP-2 MMP-9
分类号:
R336;R329.28;R392.3
文献标志码:
A
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 stimulating factor,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(tartrate-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。
Abstract:
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.

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更新日期/Last Update: 2011-07-12