生殖股神经在单侧隐睾鼠对侧睾丸损害中的作用机制

目的：探讨生殖股神经在单侧隐睾鼠模型对侧睾丸损害中的作用机制。方法：建立单侧隐睾鼠模型（21d龄），切断该侧生殖股神经，120d后观察对侧睾丸的生精细胞凋亡及细胞乳酸含量变化。结果：切断生殖股神经后对侧睾丸生精细胞凋亡减少，乳酸含量降低，乳酸含量与细胞凋亡呈正相关。结论：单侧隐睾症对侧睾丸损伤可能与其神经传导反射性血流减少引起生精细胞凋亡有关。     

小鼠单侧睾丸损伤后环孢素A对Fas系统的影响

目的 :研究昆明小鼠 (KM小鼠 )单侧注射冰乙酸致睾丸损伤后环孢素A(CsA)对对侧睾丸生精功能和Fas系统表达的影响。　方法 :6 0只KM小鼠随机分为 4组 :A组为对照组 ,B组为单侧睾丸损伤组 ,C组为单侧睾丸损伤后 6h切除损伤睾丸组 ,D组为单侧睾丸损伤后 6h内开始腹腔注射CsA组。 4周后取对侧附睾尾 ,计数精子及其活率 ,对侧睾丸作石蜡切片苏木精 伊红染色和免疫组化链霉素抗生物素蛋白过氧化物酶连结 (SP)法检测Fas和FasL的表达。　结果 :D组附睾尾精子和活率计数显著高于B组 (P <0 .0 5 ) ,D组的FasL和Fas较B组显著降低 (2 4 .3± 7.0vs37.8± 5 .8和 17.8± 4 .3vs32 .4± 3.6 ,P <0 .0 5 )。　结论 :KM小鼠单侧睾丸损伤后CsA可以通过抑制Fas和FasL的表达 ,降低生精细胞凋亡 ,维持生精功能的稳定     

单侧隐睾患儿术后青春期性发育的研究

目的　观察隐睾患儿术后青春期性发育状况。　方法　随访调查166例隐睾患儿术后青春期阴毛发育程度、阴茎长度和周径、睾丸体积和血清黄体生成素(LH)、卵泡刺激素(FSH)、睾酮(T)水平。根据术式分为单侧隐睾切除术及固定术组,根据手术年龄将固定术组分为手术年龄<5岁及≥5岁组。比较不同手术年龄、手术方式和隐睾位置对疗效的影响。　结果　单侧隐睾固定术患儿的术侧睾丸体积明显小于下降侧(15. 5±2. 4mlvs8. 1±2. 1ml)、但明显大于未治疗组的隐睾侧(4. 0±2. 0ml)。单侧隐睾固定术、切除术和正常组患儿FSH分别为(4. 5±1. 7、6. 2±1. 8、2. 4±1. 1mIU/ml。手术年龄<5岁患儿的阴毛发育程度、阴茎长度和周径、术侧睾丸体积和T明显大于≥5岁组,后者的FSH水平明显高于前者。高位型患儿术侧睾丸体积明显降低、FSH明显升高。睾丸体积与FSH呈负相关关系(r=-0.291,P<0. 05)。　结论　单侧隐睾患儿术后青春期性征发育可表现为正常,早期手术可防止隐睾和对侧正常下降睾丸生精功能的损害。     

单侧隐睾大鼠对侧睾丸的损害与Bcl-2和Bax基因表达

目的:探讨单侧隐睾大鼠对侧睾丸生精细胞凋亡与Bcl-2/Bax基因表达的关系。方法:20只健康SD雄性大鼠(22日龄)随机分成隐睾组和对照组,每组10只。通过手术建立单侧隐睾动物模型。术后90 d取对侧睾丸,采用原位缺口末端标记(TUNEL)法检测生精细胞凋亡,免疫组化SP法检测Bcl-2/Bax基因表达。结果:与对照组相比,隐睾组对侧睾丸生精细胞凋亡显著增多(P<0.01),重量显著减轻(P<0.01),Bax表达显著升高(P<0.01),Bcl-2表达显著降低(P<0.01)。凋亡细胞主要是初级精母细胞和圆形精子细胞。结论:单侧隐睾大鼠对侧睾丸的生精细胞凋亡增多与Bcl-2基因表达降低、Bax基因表达升高密切相关。细胞内Bc l-2/Bax比值是影响生精细胞凋亡的重要因素之一。     

抗氧化酶对隐睾生精细胞凋亡的影响

目的 :探讨超氧化物歧化酶 (SOD)、过氧化氢酶 (CAT)、谷胱甘肽过氧化物酶 (GSHPx)对隐睾生精细胞凋亡的影响。　方法 :4 8只未成熟SD雄性大鼠随机分为隐睾组和对照组 ,于术后第 1、3、7d采集睾丸。TUNEL法检测其生精细胞凋亡 ;化学比色法测定其SOD、CAT、GSHPx的活性和丙二醛 (MDA)含量。　结果 :术后第 7d ,与对照组相比 ,隐睾重量、SOD活性、CAT活性和SOD/ (CAT +GSHPx)比值均显著降低 (P均 <0 .0 1) ,GSHPx的活性无显著变化 (P >0 .0 5 ) ,生精细胞凋亡指数和MDA含量均显著增加 (P均 <0 .0 1)。　结论 :隐睾生精细胞的凋亡与抗氧化酶活性的降低密切相关     

单侧睾丸扭转引发对侧睾丸生精损伤免疫机制的实验研究

目的探讨单侧睾丸扭转对侧睾丸生精功能损伤的机理。方法雄性SD大鼠24只,随机分为3组,建立睾丸扭转动物模型。第1组为假手术组,第2、3组扭转左侧睾丸720°并固定维持,分别于12 h和24 h后复位固定。术后1个月留取对侧睾丸称重。应用流式细胞术(FCM)检测各组生精细胞凋亡和各级生精细胞计数,免疫组化SABC法检测抗精子抗体免疫复合物(IgG)在睾丸组织中的沉积。结果第1、2、3组对侧睾丸重量分别为(1555.73±72.34)、(1184.20±101.02)、(783.60±117.93)mg,凋亡细胞数依次为53.25±8.61、1622.00±129.31、3401.25±179.75,抗精子抗体阳性染色率为0、0.55±0.02、0.69±0.03,组间两两比较差异均有统计学意义(P<0.01)。与第1组相比,第2、3组对侧睾丸单倍体、二倍体和四倍体细胞群计数明显减少,同时可见抗精子抗体免疫复合物在睾丸组织中明显沉积,其差异均有统计学意义(P<0.01)。第2、3组间(除四倍体细胞群计数外)差异亦有统计学意义(P<0.01)。结论单侧睾丸扭转导致对侧睾丸生精细胞广泛凋亡,自体免疫反应可能在对侧睾丸生精功能损伤中起重要作用。     

隐睾及睾丸固定术后对大鼠生精能力的影响

目的 观察隐睾及睾丸固定术后对睾丸生精能力的影响.方法 通过手术方法对80只SD大鼠制作单侧隐睾模型,随机分为10组,其中4组(每组10只)于隐睾术后7、14 d切取患侧睾丸组织,6组于隐睾术后7、14d行睾丸固定术,分别于术后2、4、6周取材.将所取得的睾丸组织称重后行流式细胞仪检测其细胞凋亡率和各生精细胞百分比以及组织中B淋巴细胞/白血病-2( bcl-2)和bax基因表达量.结果 隐睾睾丸重量明显下降,隐睾组1C细胞(7d组:10.61 ±1.10,14d组:11.79 ±0.91)较对照组降低(16.48±1.60,P＜0.05)、4C细胞也明显降低,而2C细胞(7d组:40.41±2.93,14 d组:51.41±6.45)较对照组增加(30.17±3.24,P＜0.05).隐睾各组生精细胞凋亡率(7d组:14.9±1.26,14 d组:6.90±0.96)高于正常对照组(2.50±0.44,P＜0.01),而睾丸复位固定术后各组生精细胞凋亡率均下降(P＜0.05).隐睾7、14 d睾丸中bc1-2蛋白表达量(7d组:4.68±0.47;14 d组:5.66 ±0.71)较对照组(7.47±1.01)降低而bax蛋白表达量(7d组:8.27±1.08;14 d组:6.26±0.21)较对照组(5.82 ±0.47,P＜0.05)升高,睾丸固定术后各组bcl-2蛋白表达量升高而bax蛋白表达量降低(P＜0.01).结论 隐睾可以使睾丸生精细胞凋亡增加,睾丸复位固定术后,睾丸生精功能可部分或全部恢复,其恢复程度和隐睾时间长短有关;bcl-2和bax表达在生精细胞凋亡的调控中起重要作用.     

实验性隐睾大鼠的抗精子自身免疫研究

目的　探讨大鼠隐睾模型中抗精子自身免疫的发生情况及其对双侧睾丸的影响。方法　将 2 2日龄SD雄性大鼠 2 4只随机分为隐睾组和假手术组各 12只 ,在日龄 110天时采血并处死。用ELISA法检测血清抗精子抗体 (AsAb) ,免疫组化 (SABC法 )检测睾丸组织的IgG复合物 ,并对睾丸生精功能进行评价。结果　隐睾组和假手术组的血清AsAb阳性率分别为 41.7%和 0 ,差异有显著性 (P <0 .0 5 ) ;隐睾组大鼠的隐睾及对侧睾丸的生精功能评分均显著低于假手术组的手术侧及对侧睾丸 (隐睾侧P <0 .0 0 1,对侧P <0 .0 5 ) ,隐睾组大鼠的睾丸组织IgG复合物的沉积程度与生精功能呈负相关 (隐睾侧r =- 0 .860 ,P <0 .0 0 1,对侧r =- 0 .693 ,0 .0 1

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血管内皮细胞生长因子和内皮型一氧化氮合酶在大鼠单侧隐睾对侧睾丸损害中的表达

目的 探讨大鼠隐睾中内皮型一氧化氮合酶(eNOS)和血管内皮细胞生长因子(VEGF)的表达与单侧隐睾对侧睾丸损害的关系.方法 将45只雄性SD大鼠(22 日龄)随机分成单侧隐睾并生殖股神经离断组(A组)、单侧隐睾组(B组)和假手术组(C组),每组15只.动物模型建立后(65日龄),苏木素-伊红(HE)染色观察睾丸生精上皮形态,原位缺口末端标记法(TUNEL)检测对侧睾丸生精细胞的凋亡,免疫组织化学和Western blot方法检测对侧睾丸组织中eNOS和VEGF基因表达的变化.结果 B组相对于A组和C组对侧睾丸组织中生精细胞的凋亡率最高(t1=3.04,t2=3.94,t1,t2＞t28,P＜0.01),Western blot和免疫组织化学方法检测结果也显示B组eNOS和VEGF的表达含量较A组和C组都显著升高,差异有统计学意义(P＜0.01),而A组和C组的各指标差异均无统计学意义(P＞0.05).结论 eNOS和VEGF的高表达和生殖股神经在单侧隐睾对侧睾丸的损害中起重要作用,而切断生殖股神经可以阻断这一损害过程.

Abstract：

Objective To study the relationship between the expression of endothelial nitric oxide synthase (eNOS) and vascular endothelial growth factor (VEGF) with the damage of contralateral testis of unilateral cryptorchidism in the experimental rats. Methods Forty-five immature male Sprague-Dawley rats (aged 22 days) were randomly divided into the group A (unilateral cryptorchidism and the ipsilateral genitofemoral nerve division), group B (unilateral cryptorchidism), group C (sham operation), n = 15 in each group. When the rats were aged 65 days, all the rats were sacrificed and the testes were obtained. The morphological changes of the spermatogenic epithelium in the testes were observed, and the germ cell apoptosis was detected by the TdT-mediated dUTP-biotin nick end labeling (TUNEL) technique. The expression of eNOS and VEGF was detected by using Western blotting and immunohistochemistry in the testicular tissues.Results The germ cell apoptosis was increased significantly ( t1 = 3. 04, t2 = 3. 94, t1 ,t2 ＞ t28 , P ＜0. 01) , and the levels of eNOS and VEGF in the contralateral testes were also increased obviously in group B as compared with groups A and C ( P ＜ 0. 01) , but the entire indexes in groups A and C had no significant difference. Conclusion eNOS, VEGF and genitofemoral nerve play a important role in the damage of the contralateral testes of unilateral cryptorchidism, and the damage can be prevented by genitofemoral nerve division.     

大鼠单侧隐睾对侧睾丸的损害与抗氧化酶mRNA的表达

目的　从基因表达水平探讨单侧隐睾对侧睾丸损害机制。方法　 3 0只SD雄性大鼠分为对照组与隐睾组 ,每组各 15只。采用逆转录 聚合酶链反应 (RT PCR)方法检测对侧睾丸中谷胱甘肽过氧化物酶 (GSH PX)、铜 /锌超氧化物岐化酶 (Cu/Zn SOD)、过氧化氢酶 (CAT)mRNA的表达 ;化学比色法测定丙二醛 (MDA)的含量 ;生物素 dUTP/酶标亲和素法检测睾丸生殖细胞的凋亡。结果　术后 2周 ,与对照组比较 ,隐睾组GSH PX和SODmRNA表达明显下降 ,MDA和生殖细胞凋亡明显升高 (P <0 .0 1) ,而CATmRNA表达无明显改变 (P >0 .0 5 )。结论　单侧隐睾对侧睾丸存在GSH PX和SODmRNA表达降低 ,氧自由基升高和生殖细胞过度凋亡。     

Undescended testis is accompanied by calcitonin gene related peptide accumulation within the sensory nucleus of the genitofemoral nerve in trans-scrotal rats

PURPOSE: Recent data suggest that calcitonin gene related peptide (CGRP) released from the sensory branch of the genitofemoral nerve may regulate testicular descent. We studied the number of CGRP immunoreactive cells in the sensory nucleus of the genitofemoral nerve (L1 to L2 dorsal root ganglia) in cryptorchid trans-scrotal rats. Four-week-old trans-scrotal rats with unilateral undescended testis underwent bilateral genitofemoral nerve dissection and retrograde nerve labeling with the fluorescent dye 4,6-diamidino-2-phenylindole (DAPI). Animals were sacrificed 48 hours later and the L1 to L2 dorsal root ganglia were removed. Serial sections were obtained and double fluorescent labeled with antibody to CGRP. Retrograde labeled and CGRP immunoreactive cells were counted using an epi-fluorescent microscope.In the 6 male trans-scrotal rats evaluated we noted a mean plus or minus standard deviation of 1,272 +/- 98 retrograde labeled dorsal root ganglion cells ipsilateral to a fully descended testicle, including 98 +/- 34 that were also CGRP immunoreactive. On the side of the undescended testis there was a mean of 1,600 +/- 304 DAPI positive cells and 160 +/- 51 CGRP immunoreactive, DAPI labeled cells. The difference was significant (p <0.02).This study shows that in trans-scrotal rats the sensory nucleus of the genitofemoral nerve contains more CGRP immunoreactive cells ipsilateral to an undescended testis than on the contralateral side, highlighting the significance of CGRP supply through the sensory branch of the genitofemoral nerve for testicular descent.     

单侧隐睾对侧睾丸生精上皮保护作用的实验研究

目的探讨单侧隐睾鼠对侧睾丸生殖细胞和Sertoli细胞变化和还原型谷胱甘肽(GSH)对对侧睾丸的保护作用。方法30只SD雄性大鼠分为对照组(A组)、隐睾组(B组)、隐睾加GSH组(C组),每组各10只。采用化学比色法测定对侧睾丸GSH、丙二醛(MDA)含量;生物素-dUTP/酶标亲和素法检测睾丸生殖细胞的凋亡;透射电镜观察Sertoli细胞的超微结构。结果术后2周,与A组比较,B组对侧睾丸GSH明显降低,MDA和细胞凋亡明显增加(P<0.01),Ser-toli细胞线粒体和滑面内质网扩张。C组这种变化则明显减轻(P<0.01),与A组比较差异无统计学意义(P>0.05)。结论外源性GSH对单侧隐睾鼠对侧睾丸生殖细胞和Sertoli细胞有保护作用。     

The histopathology of iatrogenic cryptorchid testis: an insight into etiology

PURPOSE: Iatrogenic undescended testis may develop after inguinal hernia repair, presumably as a result of mechanical tethering of the testis or cord in scar tissue. Because some true cryptorchid testes appear to be completely descended at birth and later ascend during childhood, some iatrogenic undescended testes may be low lying undescended testes. To determine whether iatrogenic undescended testes may be unrecognized cryptorchid testes at herniorrhaphy we examined biopsies of iatrogenic undescended testes and the corresponding contralateral descended testis. MATERIALS AND METHODS: Between 1985 and 1999 bilateral testis biopsies were obtained at orchiopexy in 37 boys 1.5 to 11.8 years old who previously underwent inguinal hernia correction. Histomorphometric analysis of germ cell counts was performed on the undescended and contralateral descended testes, and compared to the count in bilateral biopsies of 37 age and position matched patients with true unilateral cryptorchidism. RESULTS: There were no significant differences in volume or total and differential germ cell counts in the undescended and contralateral descended testes in the study groups and age matched controls with primary unilateral cryptorchidism. The mean number of germ cells per tubule in the undescended testis in patients with a greater than 5-year interval from herniorrhaphy to orchiopexy was significantly decreased compared to those with an operative interval of less than 5 years (0.27 +/- 0.33 versus 0.93 +/- 1.4, p = 0.026). CONCLUSIONS: Some patients with iatrogenic undescended testis may have an unrecognized low cryptorchid testis. Careful physical examination before and after inguinal surgery is recommended. The early repair of iatrogenic undescended testis is warranted to prevent further damage.     

Ν-硝基-L-精氨酸甲酯对大鼠隐睾生殖细胞凋亡的保护作用

目的 :探讨一氧化氮合酶 (NOS)抑制剂N 硝基 L 精氨酸甲酯 (L NAME)对大鼠隐睾生殖细胞凋亡的保护作用。　方法 :用 2 2dSD雄性大鼠复制单侧隐睾模型。实验分假手术组、隐睾组、隐睾 +L NAME组 [术后腹腔注射L NAME ,10mg/(kg·d) ],每组大鼠各 10只。术后 7d ,用生物素 dUTP/酶标亲和素测定法检测睾丸生殖细胞凋亡 ,用硝酸还原酶法测定睾丸组织中N0含量 ,用化学比色法测定睾丸组织中NOS活性。　结果 :术后第 7d ,与假手术组睾丸相比 ,隐睾组睾丸发生凋亡的生殖细胞数显著增加 ,隐睾 +L NAME组睾丸发生凋亡的生殖细胞数比隐睾组显著减少 (P <0 .0 1) ,隐睾 +L NAME组睾丸组织中NO含量及NOS活性与隐睾组相比显著降低 (P<0 .0 1)。　结论 :隐睾组织中NO和NOS升高是隐睾生殖细胞凋亡增加的病理机制之一 ,L NAME通过抑制NOS活性、减少NO的产生来降低睾丸组织生殖细胞的凋亡发挥其保护作用。     

Assessment of germ cell apoptosis in cryptorchid rats

Aim: To investigate the relationship between germ cell degeneration and apoptosis in cryptorchid rats. Methods: Thirteen 21-day-old Wistar rats were made unilaterally cryptorchid by closing the left inguinal canal. At day 30 (Group 1, n=6) and day 60 (Group 2, n=7) after operation, the testes were removed for histopathological examination. The controls (n=8) were sham operated and were sacrificed at day 60. Germ cell apoptosis was assessed by means of the TUNEL method. Results: Spermatogenesis was arrested and the testicular and seminiferous tubular diameters were significantly reduced In the unilateral undescended testes (UUTs) compared with the contralateral descended testes (CDTs) and the control rats. However, atrophic changes, pathological calcification, necrosis of seminiferous tubule, and absence or sloughing of germ cells were not found in all the animals. The spermatocytes were the main type of germ cells undergoing apoptosis in all the groups. In the UUTs, there was a significant and time-depe     

N-硝基-L-精氨酸甲酯对大鼠隐睾生殖细胞凋亡的保护作用

目的探讨一氧化氮合酶(NOS)抑制剂N-硝基-L-精氨酸甲酯(L-NAME)对大鼠隐睾生殖细胞凋亡的保护作用.方法用22 d SD雄性大鼠复制单侧隐睾模型.实验分假手术组、隐睾组、隐睾+L-NAME组[术后腹腔注射L-NAME,10 mg/(kg·d)],每组大鼠各10只.术后7 d,用生物素-dUTP/酶标亲和素测定法检测睾丸生殖细胞凋亡,用硝酸还原酶法测定睾丸组织中NO含量,用化学比色法测定睾丸组织中NOS活性.结果术后第7 d,与假手术组睾丸相比,隐睾组睾丸发生凋亡的生殖细胞数显著增加,隐睾+L-NAME组睾丸发生凋亡的生殖细胞数比隐睾组显著减少(P＜0.01),隐睾+L-NAME组睾丸组织中NO含量及NOS活性与隐睾组相比显著降低(P＜0.01).结论隐睾组织中NO和NOS升高是隐睾生殖细胞凋亡增加的病理机制之一,L-NAME通过抑制NOS活性、减少NO的产生来降低睾丸组织生殖细胞的凋亡发挥其保护作用.