二花脸猪隐睾和正常睾丸组织学观察及PCNA/CD31蛋白差异表达分析  

Histological observations and differential expression analysis of PCNA/CD31 protein in the cryptorchid and normal testis of Erhualian pigs

作　　者：刘鹏刚 刘彩虹 包文斌 邵一铭 LIU Peng-gang;LIU Cai-hong;BAO Wen-bin;SHAO Yi-ming(College of Veterinary Medicine,Yangzhou University,Yangzhou 225009,China;Joint Laboratory of International Cooperation on Agriculture And Agricultural product safety,Ministry of Education,Yangzhou University,Yangzhou 225009,China;Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses,Yangzhou 225009,China;Key Laboratory for Animal Genetics,Breeding,Reproduction and Molecular Design of Jiangsu Province,College of Animal Science and Technology,Yangzhou University,Yangzhou 225009,China)

机构地区：[1]扬州大学兽医学院,江苏扬州225009 [2]扬州大学,教育部农业与农产品安全国际合作联合实验室,江苏扬州225009 [3]江苏省动物重要疫病与人兽共患病防控协同创新中心,江苏东扬州225009 [4]扬州大学动物科技学院,江苏省动物遗传育种繁殖与分子设计重点实验室,江苏扬州225009

出　　处：《中国兽医科学》2023年第3期388-395,共8页Chinese Veterinary Science

基　　金：国家自然科学基金青年基金项目(32102621);江苏高校优势学科建设工程资助项目(PAPD);2022年度扬州市软科学研究课题(2022019);扬州大学博士后项目(137070547)。

摘　　要：为探究二花脸猪隐睾组织学特点和PCNA/CD31的差异性表达,本试验以二花脸猪隐睾和正常睾丸组织作为研究对象,通过组织特殊染色观察分析猪隐睾组织学特点,同时利用蛋白免疫印迹、免疫组织化学和免疫荧光等实验,检测PCNA/CD31蛋白在隐睾和正常睾丸组织中的差异性表达,为研究隐睾的发生机制和种公猪睾丸发育的检测提供实验数据和理论依据。组织学分析结果显示,HE染色观察隐睾组织睾丸小叶界限不清、间质细胞多而散乱分布、曲精小管直径明显减小,管腔内可见大量精原细胞及支持细胞;VG和Ag NOR染色观察睾丸结缔组织和曲精小管基膜中呈红色的胶原纤维较少,且胶原纤维分布紊乱,核分裂相细胞较少。蛋白免疫印迹检测PCNA/CD31在隐睾组织中的表达均显著高于正常睾丸组织(P<0.01)。免疫组织化学结果显示,PCNA/CD31在隐睾和正常睾丸组织中都存在阳性表达,PCNA在隐睾和正常睾丸结缔组织的血管壁和间质细胞的胞质中强阳性表达,在胞核中也可见少量阳性表达;CD31表达于隐睾和正常睾丸组织间质细胞的胞质中,而在细胞膜和胞核中未见表达。结果表明,PCNA/CD31参与了睾丸间质细胞的增殖和分布、血管的生成及曲精小管的发育,是诱导猪隐睾发生的关键因子。The purpose of this study is to explore the histological characteristics of cryptorchidism and differential expression of PCNA/CD31 in the testes of Erhualian pigs.In this experiment,the cryptorchid testicular tissue of Erhualian pigs was used as the research object and differential expression of PCNA/CD31 in the cryptorchid tissue was detected through tissue special staining observation,protein immunoblotting,immunohistochemistry,and immunofluorescence.The following results were obtained:from HE staining,testicular lobules in cryptorchid tissues were poorly defined,interstitial cells were numerous and scattered,the diameter of convoluted seminiferous tubules was significantly reduced,and a large number of spermatogonia and supporting cells were visible in the lumen.vG and AgNOR staining showed that there were fewer red collagen fibers in the testicular connective tissue and basal membrane of seminiferous tubules,with a disordered distribution of collagen fibers and fewer mitotic cells.Protein immunoblotting showed no difference in the expression of PCNA/CD31 in cryptorchid tissues of different individual pigs,while expression in cryptorchid tissues was significantly higher than that in normal testicular tissues(P<0.01).Immunohistochemistry showed that PCNA/CD31 was positively expressed in both cryptorchid and normal testis.Strong positive expression of PCNA was found in the vascular wall,cytoplasm of interstitial cells,and to a lesser extent in the nucleus of cryptorchid and normal testicular connective tissues,CD31 was expressed in the cytoplasm of interstitial cells of cryptorchid and normal testis,while none in the cell membrane,and the nucleus.The results show that PCNA/CD31 is involved in the proliferation and distribution of Leydig cells,angiogenesis,and the development of convoluted seminiferous tubules,and is a key factor in the induction of cryptorchidism in male animals.

关 键 词：睾丸 猪 隐睾 增殖细胞核抗原 CD31 

分 类 号：S852.16[农业科学—基础兽医学]