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骨痂组织冰冻切片前胶原及转化生长因子－β1基因表达的原位定位 |
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史炜镔 杜宁 符诗聪 程枫 金晓龙 张凤华 杨庆铭 柴本甫
【摘要】目的观察骨痂组织中前胶原、转化生长因子－β1(transforming growth factor β1,TGF－β1)基因表达模式，分析TGF－β1在骨折愈合过程中的作用，探索骨组织冰冻切片原位杂交技术。方法采用不脱钙的大鼠骨痂组织冰冻切片进行原位杂交，观察骨痂组织中前胶原和TGF－β1基因的表达，并与先前研究作对照。结果杂交信号清晰，定位良好，特异性高。骨折第1周末，成纤维细胞的Ⅲ型前胶原基因表达占主导，Ⅰ型前胶原mRNA阳性成骨细胞也出现于膜内化骨区。TGF－β1在分化、增殖的成骨细胞以及接近成熟的软骨细胞有显著表达。骨折第2周末，Ⅱ型前胶原和TGF－β1mRNA在成熟的软骨细胞大量表达，而Ⅰ型前胶原mRNA表达也明显增加。骨折第4周末，软骨骨痂基本被骨组织替代，见散在Ⅰ型前胶原mRNA表达阳性的成骨细胞。同时证实共有表型表达的现象存在。结论实验结果与以往有关研究结果的吻合，提示TGF－β1在骨折愈合过程中，尤其在细胞分化、增殖中，起重要的调节作用。同时，也说明了此方法是一种快捷、灵敏、又不失特异性的骨组织原位杂交方法。
【关键词】骨痂冷冻切片原位杂交转化生长因子β基因表达
In Situ Localization of Procollagen and Transforming Growth Factor Gene
Expression on Cryosection of Fracture Callus
SHI Weibin,DU Ning,FU Shicong,et al.
Department of Traumatology,Ruijin Hospital,Shanghai Second Medical University,ShanghaiN 200025
【Abstract】Objective To investigate the expression mode of procollagen and transforming growth factor β1(TGF－β1)gene in fracture callus,analyze the role of TGF－β1 in fracture healing,and grope for the technique of in situ hybridization for skeletal tissue.Methods In situ localization of procollagen and TGF－β1 gen eexpression was performed on the cryosection of rat fracture callus,and the results were compared with those of earlier study.Results The hybridization signals were clear and easy to be localized with high specificity.On the seventh day after fracture,the expression of pro α 1(Ⅲ)in fibroblast and chondrocyte-like cell swa sdominant.TGF－β1 was strongly expressed in differentiating proliferating osteoblasts and premature chondrocytes.The end of second week was characterized by a marked increase in the mRNA levels of type－Ⅱprocollagen and TGF－β1 in mature chondrocytes.At the end of fourth week,the cartilaginous callus was almost replaced by the osseous tissue.Some type－I procollagen mRNA positive osteoblasts and hypertrophic ch[1] [2] 下一页
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