Toll样受体亚型在干细胞成骨过程中的作用Effect of Toll-like receptor on the ossification capacity of bone marrow mesenchymal stem cells
王晓光;陈奇;徐晓峰;张志坚;胡浪;
摘要(Abstract):
[目的]探索大鼠骨髓间充质干细胞(Bone Marrow Mesenchymal Stem Cells,BMSCs)成骨过程各阶段,不同Toll样受体(Toll Like Receptor,TLR)亚型的作用。[方法]利用贴壁培养法分离纯化SD大鼠BMSCs,传代扩增,免疫荧光分析检测细胞表面分子CD44、CD105、Vimentin的表达并进行细胞鉴定。选取活力好的第3代细胞,随机分为3组:空白组,TLR-3激活组和TLR-4激活组,均以成骨诱导培养液在相同的环境下培养。Westernblot法检测Collagen-I和Osteocalcin的表达,观察干细胞向成骨细胞(Osteoblast,OB)分化。于培养10、14 d进行碱性磷酸酶染色及茜素红染色,观察矿化结果。[结果]细胞表面抗原CD44、CD105、Vimentin呈阳性。Collagen-I和Osteocalcin的蛋白表达依次为,TLR-4激活组>TLR-3激活组>空白组。3组碱性磷酸酶染色及茜素红染色均呈阳性,TLR-3激活组强度最高,TLR-4激活组和空白组差异不明显。[结论]在BMSC成骨过程中,不同TLR亚型在不同的阶段起作用,TLR-3和TLR-4均可在BMSCs向OB分化过程中起促进作用,TLR-4的效果更明显。在OB成熟矿化为骨细胞的过程中,TLR-3起促进作用,而TLR-4则起抑制作用。
关键词(KeyWords): Toll样受体;骨髓间充质干细胞;成骨分化;矿化
基金项目(Foundation): 镇江市科技发展基金项目(编号:SH2002019)
作者(Author): 王晓光;陈奇;徐晓峰;张志坚;胡浪;
Email:
DOI:
参考文献(References):
- [1]徐晓峰,刘小平,王明伟,等.血管内皮生长因子-纳米晶胶原基骨缓释系统与人骨髓间充质干细胞的体外生物相容性[J].中国组织工程研究与临床康复,2010,25:4636-4640.
- [2]何川,杨庆铭.骨髓源性间充质干细胞的基本概念和在骨科的相关应用研究[J].中国矫形外科杂志,2005,5:389-392.
- [3]Glass GE,Chan JK,Freidin A,et al.TNF-αpromotes fracture repair by augmenting the recruitment and differentiation of muscle-derived stromal cells[J].Proc Natl Acad Sci U S A,2011,108:1585-1590.
- [4]Mountziaris PM,Mikos AG.Modulation of the inflammatory response for enhanced bone tissue regeneration[J].Tissue Eng Part B Rev,2008,14:179-186.
- [5]汤亭亭,戴尅戎,张世民,等.冻干骨对间充质干细胞成骨能力的影响[J].中国矫形外科杂志,2005,11:850-852.
- [6]Francesco L,Roberta A,Lorenzo C,et al.Toll-like receptors 3 and4 are expressed by human bone marrow-derived mesenchymal stem cells and can inhibit their t-cell modulatory activity by impairing notch signaling[J].Stem Cells,2008,26:279-289.
- [7]Waterman RS,Tomchuck SL,Henkle SL,et al.A new mesenchymal stem cell(MSC)paradigm:polarization into a pro-inflammatory MSC1 or an immunosuppressive MSC2 phenotype[J].PLoS One,2010,4:e10088.
- [8]Nam D,Mau E,Wang YF,et al.T-Lymphocytes enable osteoblast maturation via IL-17F during the early phase of fracture repair[J].PLoS One,2012,6:e40044.
- [9]Lepperdinger G.Inflammation and mesenchymal stem cell aging[J].Curr Opin Immunol,2011,23:518-524.
- [10]Wennberg C,Hessle L,Lundberg P,et al.Functional characterization of osteoblasts and osteoclasts from alkaline phosphatase knock out mice[J].J Bone Miner Res,2000,10:1879-1888.
- [11]白庆霞,杨博,张艳,等.Toll样受体与经典Wnt信号在破骨样细胞中作用机制的初步研究[J].牙体牙髓牙周病学杂志,2013,3:156-161.
- [12]温家兵.Toll样受体信号通路与骨关节炎相关性的研究进展[J].中国矫形外科杂志,2012,20:357-359.