性激素受体在子午岭黑山羊隐睾及正常睾丸中的分布比较

doi:10.11843/j.issn.0366-6964.2020.029

摘要/Abstract

摘要： 旨在研究性激素受体[雄激素受体（androgen receptor，AR）、雌激素受体（estrogen receptor，ER）]在子午岭黑山羊正常睾丸与隐睾组织中的分布与隐睾症的关系，应用免疫组织化学及免疫荧光技术方法结合形态计量学统计软件，比较了正常睾丸与隐睾的组织化学特点。特殊染色结果显示：与正常组相比，隐睾组间质组织疏松，管腔面积明显减小，糖原含量明显减少，胶原纤维及网状纤维含量增多。免疫组化及免疫荧光结果显示：1） AR在正常睾丸组Leydig细胞呈高密度强阳性表达，在各级生精细胞呈中等强度阳性表达，隐睾组中Leydig细胞及各级生精细胞表达明显减弱，且在精原细胞偶见表达；2） ER在正常睾丸Leydig细胞呈高密度强阳性表达，管周肌样细胞呈中等强度阳性表达，Sertoli细胞偶见表达，各级生精细胞无表达；3） ER在隐睾Leydig细胞、初级精母细胞、精原细胞和Sertoli细胞中均呈中等强度阳性表达；4）统计结果显示，隐睾组AR的平均光密度较正常组显著降低（P<0.05），而ER的平均光密度则显著高于正常组（P<0.01），且隐睾组AR与ER表达量比值基本接近1：1。子午岭黑山羊隐睾组织胶原纤维和网状纤维分布较正常睾丸多，生精小管基膜主要成分以中性糖蛋白为主，酸性糖蛋白含量明显降低影响精子的正常形成；隐睾组织精原细胞及Sertoli细胞ER与AR表达失常尤为明显，可为哺乳动物隐睾的相关研究提供一定参考。

关键词: 子午岭黑山羊, 雄激素受体, 雌激素受体, 睾丸, 隐睾

Abstract: To study the relationship between the distribution of sex hormone receptors (AR and ER) in normal and cryptorchid testis and cryptorchidism of the Ziwuling black goat, the histochemical characteristics of normal testis and cryptorchidism were compared by immunohistochemistry, immunofluorescence and morphometric statistical software. The results of special staining showed that compared with the normal group, the interstitial tissue of cryptorchidism group was loose, the lumen area was significantly reduced, the content of glycogen was significantly decreased, and the contents of collagen and reticular fibers were increased. The results of immunohistochemistry and immunofluorescence showed that:1) AR was strongly expressed in Leydig cells of the normal testis and moderately positive in all levels of spermatogenic cells, while the expression of Leydig cells and Spermatogenic cells in cryptorchidism group was significantly decreased and occasionally expressed in Spermatogonia. 2) ER was strongly expressed in the normal testicular Leydig cells, moderately positive in Peritubular myoid cells, occasionally expressed in Sertoli cells and not expressed in Spermatogenic cells. 3) ER was moderately positive in cryptorchidism Leydig cells, primary Spermatocytes, Spermatogonia and Sertoli cells. 4) The average optical density of AR in cryptorchidism group was significantly lower than that in the normal group, while the average optical density of ER in cryptorchidism group was significantly higher than that in the normal group. The ratio of AR to ER expression in cryptorchidism group was close to 1:1. The distribution of collagen and reticular fibers in cryptorchidism of Ziwuling black goat is more than that of the normal testis, and the main component of seminiferous tubule basement membrane is the neutral glycoprotein. The decrease of the content of acid glycoprotein may affect the normal formation of spermatozoa. The expression of ER and AR is abnormal during cryptorchidism, especially the expression of spermatogonia and Sertoli cells is opposite to that of AR, which can provide some reference for the related research of cryptorchidism in mammals.

Key words: Ziwuling black goat, androgen receptor, estrogen receptor, testis, cryptorchidism

中图分类号:

S858.277.29

王乾梅, 袁莉刚, 李承晔, 杨洪早, 陈少宇. 性激素受体在子午岭黑山羊隐睾及正常睾丸中的分布比较[J]. 畜牧兽医学报, 2020, 51(6): 1455-1465.

WANG Qianmei, YUAN Ligang, LI Chengye, YANG Hongzao, CHEN Shaoyu. Distribution of Sex Hormone Receptors in Cryptorchidism and Normal Testes of Ziwuling Black Goat[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(6): 1455-1465.

参考文献 46

[1]	HUTSON J. Testis embryology, anatomy and physiology[M]//LEDBETTER D J, JOHNSON P R V. Endocrine Surgery in Children. Berlin, Heidelberg:Springer, 2018:271-279.
[2]	KRUTSKIKH A, DE GENDT K, SHARP V, et al. Targeted inactivation of the androgen receptor gene in murine proximal epididymis causes epithelial hypotrophy and obstructive azoospermia[J]. Endocrinology, 2011, 152(2):689-696.
[3]	ZHU C F, LUONG R, ZHUO M, et al. Conditional expression of the androgen receptor induces oncogenic transformation of the mouse prostate[J]. J Biol Chem, 2011, 286(38):33478-33488.
[4]	WANG J J, LI J M, XU W, et al. Androgen promotes differentiation of PLZF+ spermatogonia pool via indirect regulatory pattern[J]. Cell Commun Signal, 2019, 17(1):57.
[5]	傅冷西, 李笃妙, 张建星, 等. 隐睾症中激素和雌雄激素受体的研究[J]. 中华小儿外科杂志, 2005, 26(4):186-188.FU L X, LI D M, ZHANG J X, et al. Androgen and estrogen receptor expressions in cryptorchidism[J]. Chinese Journal of Pediatric Surgery, 2005, 26(4):186-188. (in Chinese)
[6]	BULUN S E. Aromatase and estrogen receptor α deficiency[J]. Fertil Steril, 2014, 101(2):323-329.
[7]	COOKE P S, NANJAPPA M K, KO C, et al. Estrogens in male physiology[J]. Physiol Rev, 2017, 97(3):995-1043.
[8]	VALCARCE D G, VUELTA E, ROBLES V, et al. Paternal exposure to environmental 17-alpha-ethinylestradiol concentrations modifies testicular transcription, affecting the sperm transcript content and the offspring performance in zebrafish[J]. Aquat Toxicol, 2017, 193:18-29.
[9]	DUMASIA K, KUMAR A, DESHPANDE S, et al. Differential roles of estrogen receptors, ESR1 and ESR2, in adult rat spermatogenesis[J]. Mol Cell Endocrinol, 2016, 428:89-100.
[10]	李学勤, 詹江华. INSL-3基因突变与隐睾的研究进展[J]. 天津医药, 2009, 37(1):74-76.LI X Q, ZHAN J H. Research progress of INSL-3 gene mutation and cryptorchidism[J]. Tianjin Medical Journal, 2009, 37(1):74-76. (in Chinese)
[11]	蒋学武, 邓汪东, 胡显良, 等. 外源性雌激素对小鼠睾丸引带收缩和增殖活性的影响[J]. 中华小儿外科杂志, 2005, 26(9):488-491.JIANG X W, DENG W D, HU X L, et al. The effect of exogenous estrogens on the contraction and proliferation of gubernaculum testis in fetal mice[J]. Chinese Journal of Pediatric Surgery, 2005, 26(9):488-491. (in Chinese)
[12]	中华医学会小儿外科学分会泌尿外科学组. 隐睾诊疗专家共识[J]. 中华小儿外科杂志, 2018, 39(7):484-487.Urology Group, Pediatric surgery Branch, Chinese Medical Association. Guideline of diagnosis and treatment of cryptorchidism[J]. Chinese Journal of Pediatric Surgery, 2018, 39(7):484-487. (in Chinese)
[13]	胡强, 商希锋, 任子善, 等. 免疫组化法测定小儿隐睾睾丸周围组织AR, ER的意义[J]. 潍坊医学院学报, 2003, 25(2):99-101.HU Q, SHANG X F, REN Z S, et al. Androgen and estrogen receptors expressions detected by immunochemistry method(SP) in paratesticular tissues of children with cryptorchidism[J]. Acta Academiae Medicinae Weifang, 2003, 25(2):99-101. (in Chinese)
[14]	CARREAU S, HESS R A. Oestrogens and spermatogenesis[J]. Philos Trans Roy Soc B, 2010, 365(1546):1517-1535.
[15]	李芳芳, 戴建军, 吴彩凤, 等. 克隆猪隐睾症雌雄激素及其受体基因表达的研究[J]. 上海农业学报, 2015, 31(2):18-21.LI F F, DAI J J, WU C F, et al. Expression of testosterone, estrogen and their receptors in cryptorchidism of cloned pigs[J]. Acta Agriculturae Shanghai, 2015, 31(2):18-21. (in Chinese)
[16]	PAICK J S, PARK K, KIM S W, et al. Increased expression of hypoxia-inducible factor-1α and connective tissue growth factor accompanied by fibrosis in the rat testis of varicocele[J]. Actas Urol Esp, 2012, 36(5):282-288.
[17]	HOFLACK G, VAN DEN BROECK W, MAES D, et al. Testicular dysfunction is responsible for low sperm quality in Belgian Blue bulls[J]. Theriogenology, 2008, 69(3):323-332.
[18]	袁莉刚, 张勇, 李聪, 等. 高原地区藏绵羊与小尾寒羊睾丸细胞外基质组织分布特征比较[J]. 解剖学报, 2017, 48(2):179-186.YUAN L G, ZHANG Y, LI C, et al. Comparison of distribution characteristics of extracellular matrix components in the testis of the Tibetan sheep and the small-tail Han sheep from plateau[J]. Acta Anatomica Sinica, 2017, 48(2):179-186. (in Chinese)
[19]	陈国娟, 袁莉刚, 李聪, 等. 高原牦牛隐睾组织结构特征[J]. 畜牧兽医学报, 2015, 46(12):2282-2290.CHEN G J, YUAN L G, LI C, et al. The histologic characteristics of yak cryptorchidism[J]. Acta Veterinaria et Zootechnica Sinica, 2015, 46(12):2282-2290. (in Chinese)
[20]	SUSKIND A, HAYNER-BUCHAN A, FEUSTEL P J, et al. Fibrosis correlates with detailed histological analysis of human undescended testes[J]. BJU Int, 2008, 101(11):1441-1445.
[21]	李健, 李彧, 牛建昭. 胶原纤维与器官纤维化[J]. 解剖学报, 2003, 34(6):662-664.LI J, LI Y, NIU J Z. Review of collagen fiber and organic fibrosis[J]. Acta Anatomica Sinica, 2003, 34(6):662-664. (in Chinese)
[22]	杨勇正, 滕松山. 重复热处理对大鼠睾丸的支持细胞和间质组织的影响[J]. 解剖学报, 1984, 15(4):419-423.YANG Y Z, TENG S S. The effect of repeated heating on the sertoli cells and the interstitial tissue in the rat testis[J]. Acta Anatomica Sinica, 1984, 15(4):419-423. (in Chinese)
[23]	陈祥义, 尹晓娜, 姚秀娟, 等. 唯支持细胞综合征病理变化和发病机制[J]. 温州医学院学报, 2012, 42(5):419-423.CHEN X Y, YIN X N, YAO X J, et al. The pathology and pathogenesis of sertoli cell only syndrome[J]. Journal of Wenzhou Medical, 2012, 42(5):419-423. (in Chinese)
[24]	MOORE B C, HAMLIN H J, BOTTERI N L, et al. Posthatching development of Alligator mississippiensis ovary and testis[J]. J Morphol, 2010, 271(5):580-595.
[25]	贺晓舟, 张远强, 张金山. 大鼠睾丸生精小管上皮精子发生周期的PAS法判定[J]. 动物学杂志, 2004, 39(4):50-52.HE X Z, ZHANG Y Q, ZHANG J S. Evaluation of seminiferous epithelium stage in rat testis by PAS staining[J]. Chinese Journal of Zoology, 2004, 39(4):50-52. (in Chinese)
[26]	RATO L, ALVES M G, DIAS T R, et al. Testicular metabolic reprogramming in neonatal streptozotocin-induced type 2 diabetic rats impairs glycolytic flux and promotes glycogen synthesis[J]. J Diabetes Res, 2015, 2015:973142.
[27]	LAHERI S, MODI D, BHATT P. Extra-oviductal expression of oviductal glycoprotein 1 in mouse:detection in testis, epididymis and ovary[J]. J Biosci, 2017, 42(1):69-80.
[28]	丁允闽, 叶世隽, 陈咏梅, 等. 大鼠睾丸支持细胞硫酸糖蛋白-2mRNA水平的生后变化[J]. 解剖学报, 1995, 26(4):378-382.DING Y M, YE S J, CHEN Y M, et al. Postnatal changes of sulfated glycoprotein-2 mRNA level in rat sertoli cells[J]. Acta Anatomica Sinica, 1995, 26(4):378-382. (in Chinese)
[29]	ELADAK S, MOISON D, GUERQUIN M J, et al. Effects of environmental Bisphenol A exposures on germ cell development and Leydig cell function in the human fetal testis[J]. PLoS One, 2018, 13(1):e0191934.
[30]	SHAN L X, ZHU L J, BARDIN C W, et al. Quantitative analysis of androgen receptor messenger ribonucleic acid in developing Leydig cells and Sertoli cells by in situ hybridization[J]. Endocrinology, 1995, 136(9):3856-3862.
[31]	CHI M L, NI M, JIA Y Y, et al. Isolation of CYP17 I, 3β-HSD and AR genes from spotted sea bass (Lateolabrax maculatus) testis and their responses to hormones and salinity stimulations[J]. J Ocean Univ China, 2019, 18(2):420-430.
[32]	WELSH M, SHARPE R M, MOFFAT L, et al. Androgen action via testicular arteriole smooth muscle cells is important for leydig cell function, vasomotion and testicular fluid dynamics[J]. PLoS One, 2010, 5(10):e13632.
[33]	WANG Y, WEI Y, TANG X, et al. Association between androgen receptor polymorphic CAG and GGC repeat lengths and cryptorchidism:a meta-analysis of case-control studies[J]. J Pediatr Urol, 2018,14(5):432.e1-432.e9.
[34]	PERERA N, SZAREK M, VANNITAMBY A, et al. An immunohistochemical analysis of the effects of androgen receptor knock out on gubernacular differentiation in the mouse[J]. J Pediatr Surg, 2018, 53(9):1776-1780.
[35]	霍涌玮, 邱曙东, 程健, 等. 雄激素受体在大鼠睾丸发育过程中的表达及雄激素对其表达的调节作用[J]. 西安医科大学学报, 2001, 22(3):193-196, 220.HUO Y W, QIU S D, CHENG J, et al. Expression of androgen receptors in the developing rat testis and its regulation by androgens[J]. Journal of Xi'an Medical University, 2001, 22(3):193-196, 220. (in Chinese)
[36]	闫振龙. 牦牛睾丸促性腺激素受体和性激素受体增龄性变化与免疫调控研究[D]. 兰州:甘肃农业大学, 2014.YAN Z L. The immune regulation study of gonadotropin receptor and hornone receptor in yak testis (Bos grunniens) at different ages[D]. Lanzhou:Gansu Agricultural University, 2014. (in Chinese)
[37]	ZHAI J X, GENG X Y, DING T, et al. An increase of estrogen receptor α protein level regulates BDE-209-mediated blood-testis barrier disruption during spermatogenesis in F1 mice[J]. Environ Sci Pollut Res, 2019, 26(5):4801-4820.
[38]	WEN Q, TANG E I, LI N, et al. Regulation of blood-testis barrier (BTB) dynamics, role of actin-, and microtubule-based cytoskeletons[M]//ALVES M G, OLIVEIRA P F. Sertoli cells:Methods and protocols. New York:Humana Press, 2018:229-243.
[39]	MENG J, HOLDCRAFT R W, SHIMA J E, et al. Androgens regulate the permeability of the blood-testis barrier[J]. Proc Natl Acad Sci U S A, 2005, 102(46):16696-16700.
[40]	王瑞安, 郑平菊, 张远强, 等. 雌激素受体在小鼠睾丸表达的免疫组织化学研究[J]. 中国组织化学与细胞化学杂志, 1999, 8(4):385-388.WANG R A, ZHENG P J, ZHANG Y Q, et al. Immunohistochemical study on the expression of estrogen receptors in testes of mouse[J]. Chinese Journal of Histochemistry and Cytochemistry, 1999, 8(4):385-388. (in Chinese)
[41]	王刚, 谷守义, 陈康宁, 等. 精子发生阻滞不育患者睾丸雌激素受体α的表达[J]. 中华男科学杂志, 2011, 17(1):27-31.WANG G, GU S Y, CHEN K N, et al. Expression of estrogen receptor α in the testis of infertile men with spermatogenic arrest[J]. National Journal of Andrology, 2011, 17(1):27-31. (in Chinese)
[42]	SABERWAL G S, SHARMA M K, BALASINOR N, et al. Estrogen receptor, calcium mobilization and rat sperm motility[J]. Mol Cell Biochem, 2002, 237(1-2):11-20.
[43]	ZHOU X H, KUDO A, KAWAKAMI H, et al. Immunohistochemical localization of androgen receptor in mouse testicular germ cells during fetal and postnatal development[J]. Anat Rec, 1996, 245(3):509-518.
[44]	商希锋, 任子善. 隐睾与雄激素受体、雌激素受体关系的研究进展[J]. 潍坊医学院学报, 2003, 25(6):458-461.SHANG X F, REN Z S. Research progress on the relationship between cryptorchidism and androgen receptor and estrogen receptor[J]. Acta Academiae Medicinae Weifang, 2003, 25(6):458-461. (in Chinese)
[45]	EDDY E M, WASHBURN T F, BUNCH D O, et al. Targeted disruption of the estrogen receptor gene in male mice causes alteration of spermatogenesis and infertility[J]. Endocrinology, 1996, 137(11):4796-4805.
[46]	STRAUSS L, KALLIO J, DESAI N, et al. Increased exposure to estrogens disturbs maturation, steroidogenesis, and cholesterol homeostasis via estrogen receptor α in adult mouse Leydig cells[J]. Endocrinology, 2009, 150(6):2865-2872.