ET-1/eNOS表达差异在牦牛隐睾发生中的作用分析

doi:10.11843/j.issn.0366-6964.2024.01.020

摘要/Abstract

摘要： 旨在比较内皮素-1（endothelin-1， ET-1）及内皮型一氧化氮合酶（endothelial nitric oxide synthase， eNOS）在牦牛正常睾丸与隐睾组织中的分布，分析其表达差异在牦牛隐睾发生中的作用。本研究采集健康和病理性成年（4岁）雄性牦牛睾丸共20对，分为3组：正常组（10对）、单侧下降组（6对）及隐睾组（4对），应用H.E染色、Masson’s三色染色、Gomori’s染色结合形态计量学统计软件比较正常睾丸与隐睾的组织化学特点；通过免疫组织化学方法、免疫组织荧光技术及实时荧光定量PCR （quantitative real-time PCR， qPCR）检测ET-1和eNOS在正常睾丸与隐睾中的表达量并比较组间差异。结果表明：与正常组相比较，牦牛单侧下降组间质面积/管腔面积之比无显著性差异（P>0.05）；隐睾组管腔面积明显减小，胶原纤维和网状纤维含量增多。免疫组化和免疫荧光结果显示， ET-1表达于间质细胞、各级生精细胞，隐睾组上皮未见明显表达，主要表达于间质细胞，正常组ET-1的平均光密度与单侧下降组差异显著（P<0.05），与隐睾组差异极显著（P<0.01）；eNOS表达于生精小管、间质细胞，正常组eNOS的平均光密度与单侧下降组差异极显著（P<0.001），与隐睾组差异显著（P<0.05）。qPCR结果显示，ET-1在正常组中相对表达量显著高于单侧下降组（P<0.01）和隐睾组（P<0.01），eNOS在单侧下降组中的相对表达量相比于正常组升高（P<0.05）。高原低氧环境下，牦牛隐睾睾丸生精小管皱缩，间质细胞减少，局部分泌调节受到抑制，最终导致精子发生阻滞。ET-1和eNOS共同作用于间质细胞，在单侧下降组和隐睾组间质细胞中表达失衡，应是间质细胞及血管分布异常引起。

关键词: 牦牛, 内皮素-1, 内皮型一氧化氮合酶, 睾丸, 隐睾

Abstract: The aim of this study was to compare the distribution of endothelin-1 (ET-1) and endothelial nitric oxide synthase (eNOS) in the normal testes and cryptorchidism of yak, and analyze the role of ET-1/eNOS differences in cryptorchidism of yak. The 20 pairs of healthy and pathological adult (4-year-old) male yak testes were collected and divided into three groups: the normal group (10 pairs), the unilateral descent group (6 pairs), and the cryptorchidism group (4 pairs). The H.E, Masson’s and Gomori’s staining were used to compare the histochemical characteristics; Immunohistochemical methods, immunofluorescence techniques and quantitative real-time PCR (qPCR) were used to detect the expression levels of ET-1 and eNOS in different groups testes, and morphometric statistical software was used to compare the differences.The results showed that, compared with the normal group, there was no significant difference in the ratio of interstitial to luminal area in the unilateral descent group of yaks (P>0.05); In the cryptorchidism group, the luminal area was significantly reduced, and the content of collagen and reticular fibers were increased. Immunohistochemistry and immunofluorescence results showed: The ET-1 was expressed in leydig cells, spermatogenic cells at different stages, while mainly expressed in leydig cells and no significant expression was observed in the cryptorchidic epithelium, the difference in average optical density of ET-1 between the normal and unilateral descent groups was significant (P<0.05), also the difference between the normal and cryptorchidism groups was extremely significant (P<0.01). The eNOS was expressed in seminiferous tubules and leydig cells. Compared with the normal group, the difference in eNOS expression was extremely significant in the unilateral descent group (P<0.001) and significant in the cryptorchidism group (P<0.05). qPCR results showed that the relative expression of ET-1 in normal group was significantly higher than that in unilateral descent and cryptorchidism group (P<0.01), the relative expression of eNOS in unilateral descent group was higher than that in normal group (P<0.05). Yaks living in a hypoxic environment on the plateau, the seminiferous tubules are atrophied and interstitial cells decreased in the cryptorchidism testes of yak, ultimately leading to the local secretion regulation inhibited and sperm development obstructed. ET-1 and eNOS are co-located in leydig cells, the expression imbalance in interstitial cells in the unilateral descent group and cryptorchidism group should be caused by abnormal distribution in interstitial cells and blood vessels.

Key words: yak, endothelin-1, endothelial nitric oxide synthase, testis, cryptorchidism

中图分类号:

S823.85

张颖, 袁莉刚, 陈国娟, 张芳, 杨大鹏. ET-1/eNOS表达差异在牦牛隐睾发生中的作用分析[J]. 畜牧兽医学报, 2024, 55(1): 207-217.

ZHANG Ying, YUAN Ligang, CHEN Guojuan, ZHANG Fang, YANG Dapeng. Analysis of Differential Expression of ET-1/eNOS in the Development of Cryptorchidism in Yak[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 207-217.

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