犬隐睾及睾丸肿瘤中PGP9.5和NPY的分布比较

doi:10.16431/j.cnki.1671-7236.2020.09.006

摘要/Abstract

摘要： 试验旨在分析肽能神经递质蛋白基因产物9.5（protein gene product 9.5，PGP9.5）和神经肽Y（neuropeptide Y，NPY）在犬隐睾及睾丸肿瘤中的分布和表达，并与同年龄正常睾丸组织进行比较，为认识犬睾丸肿瘤恶变临床诊断提供参考。应用HE染色、Masson三色染色、Gomori银浸染、甲苯胺蓝染色观察各组织中网状纤维、胶原纤维及肥大细胞等组织特征，采用免疫组织化学SP法及免疫荧光法结合IPP统计分析PGP9.5和NPY在组织中的表达及定位。结果显示，正常犬睾丸生精上皮由4~7层生精细胞及Sertoli细胞构成，间质组织胶原纤维和网状纤维分布稀疏。隐睾生精小管基底膜胶原纤维厚度增加，Sertoli细胞核浓缩位于生精小管基底，间质网状纤维增多。睾丸肿瘤组织结构不清晰，胶原纤维和网状纤维无规则分布，肥大细胞较正常组及隐睾组显著增多。免疫荧光定位表明，PGP9.5在正常睾丸Leydig细胞中呈中等阳性表达，生精细胞中无明显表达；隐睾Leydig细胞及生精细胞中呈强阳性表达；睾丸肿瘤中偶有表达。NPY在正常睾丸Leydig细胞中偶见阳性表达，生精细胞中无表达；隐睾Leydig细胞及生精上皮中无表达，间质小血管管壁呈高密度强阳性表达；睾丸肿瘤组织中无明显表达。免疫组化统计表明，睾丸肿瘤组织中PGP9.5和NPY较正常组极显著降低（P<0.01），隐睾组PGP9.5和NPY表达显著或极显著增加（P<0.05；P<0.01）。因此，犬隐睾时PGP9.5及NPY的表达增高，提示犬隐睾时已有发展为肿瘤的趋势，且与肿瘤恶变程度相关。

关键词: 犬; 隐睾; 睾丸肿瘤; 蛋白基因产物9.5; 神经肽Y; 免疫组织化学

Abstract: The purpose of this study was to analyze the distribution and expression of peptidergic neurotransmitters protein gene product 9.5 (PGP9.5) and neuropeptide Y (NPY) in cryptorchidism and testicular tumors of dogs,compare them with normal testicular tissues of the same age,and provide reference for clinical diagnosis of malignant transformation in testicular tumors of dogs.HE staing,Masson trichrome staining,Gomori silver staining and toluidine blue staining were used to observe the tissue characteristics of reticular fibers,collagen fibers and mast cells.Immunohistochemical SP method and immunofluorescence combined with IPP were used to analyze the expression and localization of PGP9.5 and NPY in tissues.The results showed that the seminiferous epithelium of normal dog testis was composed of 4-7 layers of spermatogenic cells and Sertoli cells,and the distribution of collagen fibers and reticular fibers in interstitial tissue was sparse.The thickness of collagen fibers in the basement membrane of cryptorchidism seminiferous tubules increased,the nucleus of Sertoli concentrated at the base of seminiferous tubules,and the interstitial reticular fibers increased.The tissue structure of testicular tumor was unclear,collagen fibers and reticular fibers were irregularly distributed,and mast cells increased significantly compared with normal and cryptorchid groups.The immunofluorescence results showed that PGP9.5 was moderately positive in Leydig cells of normal testis,no significant expression in spermatogenic cells,strongly positive in Leydig cells and spermatogenic cells of cryptorchidism,and occasional expression in testicular tumors.NPY was occasionally expressed in normal testicular Leydig cells,but not in spermatogenic cells,strong positive expression in Leydig cells and seminiferous epithelium of cryptorchidism,high density and strong positive expression in interstitial vessels,and no obvious expression in testicular tumors.Immunohistochemical statistics showed that the expression of PGP9.5 and NPY in testicular tumor tissue were extremely significantly lower than that in normal group (P<0.01),while the expression of PGP9.5 and NPY in cryptorchidism group were significantly or extremely significantly increased (P<0.05;P<0.01).Therefore,the expression of PGP9.5 and NPY in cryptorchidism of dogs was increased suggesting that the cryptorchidism of dogs had a tendency to develop into a tumor,and was related to the degree of malignant transformation of tumor.

Key words: dogs; cryptorchidism; testicular tumor; PGP9.5; NPY; immunohistochemistry

中图分类号:

S814.4

任建明, 魏彦明, 袁莉刚, 王乾梅, 李承晔. 犬隐睾及睾丸肿瘤中PGP9.5和NPY的分布比较[J]. 中国畜牧兽医, 2020, 47(9): 2741-2750.

REN Jianming, WEI Yanming, YUAN Ligang, WANG Qianmei, LI Chengye. Comparison of the Distribution of PGP9.5 and NPY in Cryptorchidism and Testicular Tumors in Dogs[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(9): 2741-2750.

参考文献

[1] 惠风和畅.濒临消失的物种:中华田园犬[J].环境,2011,10:28-31. HUI F H C.Endangered species:Chinese garden dog[J].Environment,2011,10:28-31.(in Chinese)
[2] URESHINO R P,COSTA A J,ERUSES A G,et al.Effects of aging in the striatum and substantia nigra of a parkinson's disease animal model[J].Toxicologic Pathology,2018,46(3):348-358.
[3] 常宏建,任晓丽,张佩,等.犬常见肿瘤流行病学调查及组织病理学诊断[J].中国畜牧兽医,2018,45(5):1417-1425. CHANG H J,REN X L,ZHANG P,et al.Epidemiological investigation and histopathological diagnosis of common tumors in dogs[J].China Animal Husbandry & Veterinary Medicine,2018,45(5):1417-1425.(in Chinese)
[4] 沈清霞,王亚辰,郭苓,等.南京地区宠物犬肿瘤临床病理学诊断与发病分析[J].畜牧与兽医,2017,49(1):103-109. SHEN Q X,WANG Y C,GUO L,et al.Clinic histopathological diagnosis and carcinogenic factors analyses of tumors in pet dogs in Nanjing area[J].Animal Husbandry & Veterinary Medicine,2017,49(1):103-109.(in Chinese)
[5] 孙欣,宋志琦,刘春法,等.犬睾丸原发性特有肿瘤的病理学观察与分析[J].实验动物科学,2016,33(3):1-3. SUN X,SONG Z Q,LIU C F,et al.Pathological observation and analysis of specific dog primary testicular tumor[J].Laboratory Animal Science,2016,33(3):1-3.(in Chinese)
[6] 黄柏,袁金刚.犬腹股沟隐睾支持细胞瘤的诊疗[J].畜牧兽医科技信息,2012,93(2):110-111. HUANG B,YUAN J G.Diagnosis and treatment of inguinal cryptorchidism Sertoli cell tumor in dogs[J].Chinese Journal of Animal Husbandry and Veterinary Medicine,2012,93(2):110-111.(in Chinese)
[7] 陈健文,罗国雄,张瑜,等.10例隐睾继发精原细胞瘤的诊疗分析[J].微创泌尿外科杂志,2016,5(3):183-186. CHEN J W,LUO G X,ZHANG Y,et al.Clinical analysis of 10 cases of seminoma arising from cryptorchidism[J].Journal of Minimally Invasive Urology,2016,5(3):183-186.(in Chinese)
[8] XING J S,BAI Z M.Is testicular dysgenesis syndrome a genetic,endocrine,or environmental disease,or an unexplained reproductive disorder?[J].Life Sciences,2017,194(1):120-129.
[9] TAINIO H.Peptideric innervation of the human testis and epididymis[J].Acta Histochemica,1994,96(4);415-420.
[10] 袁莉刚,曲亚玲,谷来凤,等.不同年龄牦牛睾丸蛋白基因产物9.5和神经肽Y的分布比较[J].解剖学报,2015,46(2):232-237. YUAN L G,QU Y L,GU L F,et al.Distribution of protein gene product 9.5 and neuropeptide Y in yak testis at different ages[J].Acta Anatomica Sinica,2015,46(2):232-237.(in Chinese)
[11] 宗益平,顾民,龚永光,等.PGP9.5和NPY肽能神经在成人睾丸组织中的分布[J].南京医科大学学报(自然科学版),2008,28(3):327-329. ZONG Y P,GU M,GONG Y G,et al.Distribution of PGP9.5 and NPY immunoreactive peptidergic nerves in adult human testes[J].Acta Universitatis Medicalis Nanjing (Natural Science),2008,28(3):327-329.(in Chinese)
[12] 袁莉刚,闫振龙,陶金忠,等.PGP9.5和神经肽Y在双峰驼正常睾丸和隐睾的分布比较[J].畜牧兽医学报,2015,46(2):303-308. YUAN L G,YAN Z L,TAO J Z,et al.The distribution of PGP9.5 and NPY in the normal testis and cryptorchidism of bactrian camel[J].Acta Veteri-naria et Zootechnica Sinica,2015,46(2):303-308.(in Chinese)
[13] 袁莉刚,曲亚玲,鲁玉荣,等.不同月龄高原型藏绵羊睾丸组织中蛋白基因产物9.5和神经肽Y的分布比较[J].畜牧兽医学报,2016,47(1):71-78. YUAN L G,QU Y L,LU Y R,et al.Distribution of protein gene product 9.5 and neuropeptide Y in Tibetan plateau sheep testis at different ages[J].Acta Veterinaria et Zootechnica Sinica,2016,47(1):71-78.(in Chinese)
[14] SAINSBURY A,SCHWARZER C,COUZENS M,et al.Y4 receptor knockout rescues fertility in ob/ob mice[J].Genes & Development,2002,16(9):1077-1088.
[15] ANDREA E,DHANOOP M A,PRAVEENN K,et al.Altered expression and localization of tumor suppressive E3 ubiquitin ligase SMURF2 in human prostate and breast cancer[J].Cancers,2019,11(4):556.
[16] 张弦,陶凯雄,王国斌.MMP-2与PGP9.5在胃癌组织中的表达及其与肿瘤侵袭转移的相关性研究[J].实用癌症杂志,2006,21(4):363-365. ZHANG X,TAO K X,WANG G B.Expression of MMP-2 and PGP9.5 in gastric cancer and their relationship in tumor invasion and metastasis[J].The Practical Journal of Cancer,2006,21(4):363-365.(in Chinese)
[17] TILAN J,KITLINSKA J.Neuropeptide Y (NPY) in tumor growth and progression:Lessons learned from pediatric oncology[J].Neuropeptides,2016,55:55-66.
[18] 宋翠萍.动物隐睾模型与不育研究进展[J].实用儿科临床杂志,2009,24(23):1790-1792. SONG C P.Study progress of cryptorchidism in experimental animal models and infertility[J].Journal of Applied Clinical Pediatrics,2009,24(23):1790-1792.(in Chinese)
[19] 谢文辉,徐耀基.一例犬腹腔隐睾肿瘤及扭转的探讨[J].广东畜牧兽医科技,2017,42(4):29-31. XIE W H,XU Y J.Study on a case of abdominal cryptorchidism tumor and torsion in dogs[J].Guangdong Journal of Animal and Veterinary Science,2017,42(4):29-31.(in Chinese)
[20] 闵志雪,王朝夫,陶菁,等.睾丸支持细胞瘤8例临床病理观察[J].诊断病理学杂志,2019,26(4):331-334. MIN Z X,WANG Z F,TAO J,et al.Testis sertoli cell tumor:A clinicopathologic analysis of eight cases[J].Chinese Journal of Diagnostic Pathology,2019,26(4):331-334.(in Chinese)
[21] EMERSON R E,ULBRIGHT T M.The use of immunohistochemistry in the differential diagnosis of tumors of the testis and paratestis[J].Seminars in Diagnostic Pathology,2005,22(1):33-50.
[22] 牛云超,李雪成,刘伟,等.一例犬睾丸支持细胞瘤的病理学观察[J].黑龙江畜牧兽医,2018,20:213-214. NIU Y C,LI X C,LIU W,et al.Pathological observation of a case of testicular sertolioma in dogs[J].Heilongjiang Animal Science and Veterinary Medicine,2018,20:213-214.(in Chinese)
[23] HAMADA S,MASAMUNE A.Elucidating the link between collagen and pancreatic cancer:What's next?[J].Expert Review of Gastroenterology & Hepatology,2018,12(4):315-317.
[24] XIE S A,ZHANG T,WANG J,et al.Matrix stiffness determines the phenotype of vascular smooth muscle cell in vitro and in vivo:Role of DNA methyltrans-ferase 1[J].Biomaterials,2017,155(1):203-216.
[25] 李承晔,郭延生,袁莉刚,等.老龄哈多利系博美犬睾丸组织结构分析[J].中国畜牧兽医,2019,46(11):3198-3207. LI C Y,GUO Y S,YUAN L G,et al.Histological characteristics analysis of aging hadley pomeranian testis[J].China Animal Husbandry & Veterinary Medicine,2019,46(11):3198-3207.(in Chinese)
[26] 王晓燕,高俊岩,王艳霞.网状纤维染色在临床病理诊断中的应用及技术[J].中国医药导报,2016,13(9):72-75. WANG X Y,GAO J Y,WANG Y X,et al.Introduction and role of reticular fiber staining in pathological diagnosis of tumors[J].China Medical Herald,2016,13(9):72-75.(in Chinese)
[27] RUOSS S,HARTMANN T,CAUGHEY G H.Mast cell tryptase is a mitogen for cultured fibroblasts[J].The Journal of Clinical Investigation,1991,88(2):493-499.
[28] RAY A,LEE O,WIN Z,et al.Anisotropic forces from spatially constrained focal adhesions mediate contact guidance directed cell migration[J].Nature Communications,2017,8:14923.
[29] 曾国华,梅骅,吴开俊.先天性双侧输精管缺如患者睾丸超微结构的改变[J].中华泌尿外科杂志,2001,22(12):759-761. ZENG G H,MEI H,WU K J,et al.Study of testicular ultrastructure in congenital bilateral absence of vas deferens[J].Chinese Journal of Urology,2001,22(12):759-761.(in Chinese)
[30] MALTBY S,KHAZZAIE K,MCNAGNY K M.Mast cells in tumor growth:Angiogenesis,tissue remodelling and immune-modulation[J].Biochimica et Biophysica Acta,2009,1796(1):19-26.
[31] WROBEL K H,BRANDL B.The autonomous innervation of the porcine testis in the period from birth to adulthood[J].Anatomischer Anzeiger:Official Organ of the Anatomische Gesellschaft,1998,180(2):145-156.
[32] WEN W,LIU G,JIN K,et al.TGF-β1 induces PGP9.5 expression in CAFs to promote the growth of colorectal cancer cells[J].Oncology Reports,2017,37(1):115-122.