老龄哈多利系博美犬睾丸组织结构分析

为比较哈多利系博美犬老龄与青年阶段睾丸的组织结构差异及相关蛋白分布特征，探索博美犬不同年龄阶段睾丸组织结构变化及其对生殖功能的影响，本试验应用特殊染色、免疫组织化学法结合免疫荧光观察比较青年与老龄博美犬睾丸组织化学特点，并用IPP图像分析软件进行定量分析。结果显示，与青年博美犬相比，老龄博美犬生精上皮层数与厚度降低，Leydig细胞数及Sertoli细胞数显著增加（P<0.05），生精小管基底膜及血管管壁层胶原纤维与网状纤维含量明显增加。免疫组织化学结果显示，老龄博美犬睾丸细胞外基质（exreacellular matrix，ECM）相关蛋白Ⅳ型胶原（collage，Col Ⅳ）、硫酸乙酰肝素蛋白多糖（heparan sulfate proteoglycan，HSPG）含量极显著低于青年博美犬（P<0.01），层黏连蛋白（laminin，LN）含量显著低于青年博美犬（P<0.05）。免疫荧光结果显示，Col Ⅳ、HSPG和LN主要在Leydig细胞及Sertoli细胞强表达。因此，老龄博美犬胶原纤维与网状纤维含量增加，Leydig细胞数及Sertoli细胞数增多可能与其生精功能的维持相关，其生精功能的下降与Col Ⅳ和HSPG的变化密切相关。

Histological Characteristics Analysis of Aging Hadley Pomeranian Testis

This study was conducted to characterize the histologic structure of testicular tissue and the distribution of related protein between aging and young adult Hadley Pomeranian,and analyze the variation at different ages and influence on reproductive function.The testicular histochemical characteristics of young adult and agin Hadley Pomeranian were compared by special staining,immunohistochemistry and immunofluorescence.The quantitative analysis was carried out by IPP image analysis software.The results showed that compared with the young adult Pomeranian testis,the number and thickness of spermatogenic epithelial of the aging Pomeranian were decreased,and the number of Leydig cells and Sertoli cells were significantly increased (P<0.05).The basal layer of the seminiferous tubules and vascular wall were increased significantly.The results of immunohistochemistry showed that compared with young adult Pomeranian,the content of Col Ⅳ and HSPG in the aging Pomeranian testicular were extremely significantly decreased (P<0.01),the LN significantly decreased (P<0.05).Immunofluorescence results showed that Col Ⅳ,HSPG and LN were strongly expressed in Leydig cells and Sertoli cells.Therefore,the increased collagen and reticular fibers and the number of Leydig cells and Sertoli cells might be closely related to the spermatogenic function maintenance of the aging Pomeranian,and especially related to the Col Ⅳ and HSPG expression.