大鼠骨髓间充质干细胞成骨分化过程中降钙素基因相关肽受体的表达Detection of CGRP receptor in rat bone marrow mesenchymal stem cells and its induced osteoblasts
王钊;金丹;
摘要(Abstract):
[目的]探讨大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)向成骨细胞分化过程中降钙素基因相关肽(calcitonin gene-related peptide,CGRP)受体的表达变化。[方法]采用全骨髓法体外分离、培养大鼠骨髓间充质干细胞,分为诱导成骨组与未诱导组,在传代培养的不同时期(1、2、3周),采用免疫细胞化学、Western Blot对骨髓间充质干细胞的成骨诱导分化进行鉴定,采用RT-PCR、Western Blot检测各组细胞CGRP受体mRNA、蛋白的表达情况。[结果]RT-PCR结果显示同一时间点诱导组CGRP受体mRNA表达量高于未诱导组,诱导组CGRP受体mRNA表达以时间依赖性的方式不断增高;Western Blot结果显示同一时间点诱导组CGRP受体的蛋白水平高于未诱导组,诱导组CGRP受体蛋白水平以时间依赖性的方式增高。[结论]本实验从mRNA、蛋白水平证实大鼠骨髓间充质干细胞表达CGRP受体,随着成骨诱导的不断进行,CGRP的表达量随之上升。
关键词(KeyWords): CGRP;CGRP受体;骨髓间充质干细胞;成骨细胞
基金项目(Foundation): 国家自然科学基金项目(编号:30872638);; 教育部霍英东高等院校青年教师基金(编号:111043)
作者(Author): 王钊;金丹;
Email:
DOI:
参考文献(References):
- [1]张元平,崔继秀,裴国献,等.神经化组织工程骨构建的初步观察[J].中华创伤骨科杂志,2005,7:55-56.
- [2]韩庆林,苟三怀,王琪.降钙基因相关肽(CGRP)调控成骨细胞功能研究进展[J].中国矫形外科杂志,2005,13:544-545.
- [3]Poyner DR,Sexton PM,Marshall I,et al.The mammalian calcitoningene-related peptides,adrenomedullin,amylin,and calcitonin re-ceptors[J].Pharmacol Rev,2002,54:233-246.
- [4]Qi T,Christopoulos G,Bailey RJ,et al.Identification of N-terminalreceptor activity-modifying protein residues important for calcitoningene-related peptide,adrenomedullin,and amylin receptor function[J].Mol Pharmacol,2008,74:1059-1071.
- [5]Lerner UH.Deletions of the genes encoding catcitonin/α-CGRP,amylin and calcitonin receptor have given new and unexpected in-sights into the function of calcitonin receptors and calcitonin receptor-like receptors in bone[J].J Musculoskelet Neuronal Interact,2006,6:87-95.
- [6]Granholm S,Lundberg P,Lerner UH.Expression of the calcitoninreceptor,calcitonin receptor-like receptor and receptor activity modif-ying proteins during osteoclast differentiation[J].J Cell Biochem,2008,104:920-933.
- [7]马文辉,时述山,李亚非,等.神经肽对人成骨细胞生物学影响机理的研究[J].中国矫形外科杂志,2008,11:1091-1095.
- [8]徐格,吴雪晖,许建中,等.骨髓间充质干细胞表面CGRP受体存在的研究[J].华南国防医学杂志,2008,22:15-17.
- [9]Bjurholm A,Kreicbergs A,Brodin E,et al.Substance P-andCGRP-immunoreactive nerves in bone[J].Peptides,1988,9:165-171.
- [10]Hill EL,Elde R.Distribution of CGRP,VIP,D beta H,SP,andNPY-immunoreactive nerves in the periosteum of the rat[J].CellTissue Res,1991,264:469-480.
- [11]Danielson P,Alfredson H,Forsgren S.Distribution of general(PGP9.5)and sensory(substance P/CGRP)innervations in the humanpatellar tendon[J].Knee Surg Sports Traumatol Arthrosc,2006,14:125-132.
- [12]Li J,Kreicbergs A,Bergstr m J,et al.Site-specific CGRP innerva-tion coincides with bone formation during fracture lealing and model-ing:a study in rat angulated tibia[J].J Orthip Res,2007,25:1204-1212.
- [13]覃俊君,王簕,陈思圆,等.血管束、感觉神经束植入组织工程骨降钙素基因相关肽和受体的时空分布[J].中华创伤骨科杂志,2009,8:742-746.