肌糖蛋白TN-C促进人骨髓基质干细胞向成骨细胞的分化  

Tenascin-C induces the differentiation of human bone marrow stromal cells into osteoblasts

作　　者：曹玉净[1] 吕秋霞[1] 

机构地区：[1]河南省中医院创伤骨科,河南郑州450002

出　　处：《基础医学与临床》2014年第11期1543-1547,共5页Basic and Clinical Medicine

摘　　要：目的 研究肌糖蛋白（Tenascin-c,TN-C）对人骨髓基质干细胞向成骨细胞分化的诱导作用,为TN-C在骨折愈合中的应用提供理论依据.方法 用全骨髓培养法获取人骨髓基质干细胞（hBMSCs）,第6～10代的细胞用于本研究.体外培养的hBMSCs用自行制备的不同浓度重组TN-C （rTN-C）蛋白处理（0、1、10、50和100 nmol/L）,于处理后不同时间进行细胞茜素红染色,检测细胞的矿化;或于不同时间收集细胞总RNA进行RT-PCR检测碱性磷酸酶（ALP）和骨钙素（OCN） mRNA水平的变化;同时利用ALP检测试剂盒和骨钙素放射性免疫试剂盒分别检测ALP活性和细胞培养基中OCN含量.结果 1）rTN-C处理hBMSCs 10 d后,rTN-C呈剂量依赖性促进成骨细胞的矿化（P＜0.05）;2） rTN-C处理hBMSCs后,细胞ALP活性及培养基上清中OCN含量显著高于未处理对照组（P＜0.05）.结论 肌糖蛋白对人BMSC向成骨细胞的分化有一定的促进作用.Objective To investigate the induction effect of Tenascin-c （TN-C） on differentiation of human bone marrow stromal cells （hBMSCs） into osteoblasts.Methods hBMSCs were obtained by the whole bone marrow culture method,and cells of passages 6 ～ 10 were used for all experiments.Human BMSCs were treated with different concentration of recombinant TN-C （0,1,10,50,100 nmol/L） and osteogenic differentiation was detected by staining with Alizarin Red S.RT-PCR was performed to detect the mRNA levels of alkaline phosphatase （ALP） and osteocalcin （OCN）,ALP detection kits were used for the detection of ALP activity,and OCN radioactive immune kit was used for the determination of OCN content.Results 1） Alizarin red staining showed that direct application of exogenous rTN-C dose-dependently triggered the differentiation and mineralization of osteoblasts; 2） rTN-C treatment significantly increased the ALP avtivity and OCN content,compared with control group.Conclusions TN-C induces the differentiation of human bone marrow stromal cells （hBMSCs） into osteoblasts.

关 键 词：肌糖蛋白 人骨髓基质干细胞 成骨细胞 碱性磷酸酶 骨钙素 

分 类 号：R683[医药卫生—骨科学]