Ihh和PTHrP信号分子在猪跟腱止点的表达及意义

Expression of Ihh and PTHrP in Entheses of Achilles tendon of mini pigs

目的观察猪跟腱止点各层组织结构特点及Ⅰ型胶原(ColⅠ)、Ⅱ型胶原(ColⅡ)、Ⅹ型胶原(ColⅩ)、纤维软骨层细胞外基质以及印度豪猪蛋白(Ihh)和甲状旁腺激素相关蛋白(PTHrP)在跟腱止点中的表达分布和定位情况。方法根据跟腱止点的组织结构层次的不同,利用HE染色、番红O/固绿染色、天狼猩红染色以及免疫组织化学技术,分别观察跟腱止点各层组织结构特点,通过免疫组织化学染色观察ColⅠ、ColⅡ、ColⅩ、纤维软骨层细胞外基质以及Ihh和PTHr P在跟腱止点中的表达分布和定位情况。结果显微镜下可清晰观察到猪跟腱止点解剖层次,主要分为4个层次,波浪状的腱纤维、纤维软骨层(也称非钙化软骨层)、钙化软骨层和骨。跟腱止点组织Ⅰ、Ⅱ型胶原免疫组织化学染色证实纤维软骨层细胞外基质主要由ColⅠ、ColⅡ构成。跟腱止点组织中Ihh和PTHrP免疫组织化学染色呈阳性表现,Ihh和PTHrP阳性区域主要集中在跟腱止点的纤维软骨层和钙化层。同时可以观察到在肥大软骨细胞细胞浆中出现浓聚现象。结论跟腱止点纤维软骨层由纤维软骨细胞构成,是跟腱止点的重要结构,Ihh和PTHrP在跟腱止点纤维软骨层和钙化层的定位表达,提示两者在跟腱止点的形成过程发挥重要的调节作用。

Objective To observe each layer structure and the expression and localization of Col Ⅰ, Col Ⅱ,ColⅩ, Ihh and PTHrP in the Entheses of Achilles tendon of mini pigs. Methods According to the organizational structure of the Entheses,HE staining, Alcian blue staining and Sirius red staining were used to characterize the structure of the entheses. Col Ⅰ, ColⅡ,Col Ⅹ Immunohistochemical staining were used to observe the cartilage extracellular matrix composition. Ihh and PTHr P Immunohistochemical staining were used to observe expression and localization in the entheses. Results The entheses anatomical layer was clear, mainly divided into four layers, fibers tendon, fibrous cartilage layer（also called the Un-calcified cartilage layer）, calcified cartilage layer and bone. Extracellular matrix of entheses of Achilles tendon was made up of collagen type Ⅰ, Ⅱ. Ihh and PTHrP immunohistochemical staining was positive, Ihh and PTHrP positive area mainly concentrated in the fibrous cartilage layer and calcification of the Entheses. Hypertrophy of cartilage cell could be observed in cytoplasm.Conclusion The entheses fiber cartilage layer is made up of fibrochondrocytes, it is the important structure of the entheses,Ihh and PTHrP positioning expressed in the fibrous cartilage laye and Un-calcified cartilage layer prompt that they play an important role in regulating in the formation process of entheses.