降钙素相关肽诱导SD大鼠骨髓间充质干细胞成骨分化的机制  

Effect of calcitonin gene-related peptide on bone marrow-derived mesenchymal stem cells and osteogenic differentiation in rats

作　　者：张振宇[1] 韩一生[1] 毕龙[1] 王树森[1] 宋扬[1] 侯巍[1] 黄兆松[1] 

机构地区：[1]第四军医大学西京医院骨科,陕西西安710032

出　　处：《现代生物医学进展》2010年第24期4606-4611,共6页Progress in Modern Biomedicine

基　　金：国家自然科学基金资助项目(30872634)

摘　　要：目的:研究外源性降钙素基因相关肽(calcitonin gene-relate peptide,CGRP)对SD大鼠骨髓来源间充质干细胞(BMSCs)增殖和成骨分化功能的影响。方法:采用贴壁法分离骨髓间充质干细胞,扩增传代至第三代,根据分组,培养体系中添加含不同浓度(10-11～10-6mol/L)CGRP的条件培养液,WST-1法检测细胞增殖能力;碱性磷酸酶染色及钙结节染色法观察CGRP诱导BMSCs向成骨细胞分化、矿化的效果。采用RT-PCR方法检测碱性磷酸酶(ALP)、I型胶原(COLL-I)、BMP-2、RunX2、骨粘连蛋白(Osteonectin,ON)等成骨相关细胞因子mRNA的表达。结果:增殖率测定CGRP组各浓度均较对照组增加,且呈剂量依赖关系,CGRP浓度大于1×10-10mol/L时差异有显著性(P<0.05);碱磷酶染色与钙结节染色结果显示,CGRP组均有阳性显色,对照组无显色或显色不明显。CGRP组的细胞因子表达较对照组显著升高(P<0.05)。结论:适当浓度的CGRP能够直接促进体外培养的BMSCs增殖,并可短期内诱导其在向成骨细胞分化。CGRP可能在骨修复及骨重建中发挥重要的作用。Objective： To investigate the effects of calcitonin gene-relate peptide on the proliferation and differentiation of rat bone marrow stromal cells（BMSCs）.Methods： BMSCs were isolated from rats by density gradient centrifugation and attachment method.Different dose of calcitonin gene-relate peptide（1×10-11～1×10-6mol/L） were added to the culture media of the third passage BMSCs for the observation of proliferation（OD value at wavelength 450nm）,differentiation（ALP staining） and mineralization（The cells were stained with alizarin red for the observation of calcium nodule） of the BMSCs.RT-PCR was used to detecte the expression of ALP,Collagen I,BMP-2,RunX2 and ON.Results： The OD values increased in a dose-dependent manner（p0.05）;The ALP staining was positive.Calcium nodule formation was demonstrated by alizarin red staining.Conclusion The expressions of cytokines all increased.Conclusions： The proliferation,differentiation and mineralization of the BMSCs are promoted by appropriate concentration of CGRP directly,which may play an important role in bone repair and reconstruction.

关 键 词：降钙素基因相关肽(CGRP) 干细胞 增殖 分化 

分 类 号：Q95-3[生物学—动物学] R683[医药卫生—骨科学]