单侧隐睾大鼠生殖股神经在对侧睾丸损害中的作用机制研究

目的 :探讨单侧隐睾大鼠模型生殖股神经在对侧睾丸损害中的作用机制。　方法 :建立单侧隐睾大鼠模型 (2 1d龄 ) ,切断该侧生殖股神经 ,12 0d后观察对侧睾丸的生精细胞凋亡变化及组织乳酸含量变化。　结果 :切断生殖股神经后 ,对侧睾丸生精细胞凋亡为 (5 .76± 0 .76 ) % ,与对照组 (17.2 8± 1.36 ) %相比明显减少 (P <0 .0 5 ) ;乳酸含量也由 (2 .19± 0 .2 4 )mmol/L下降为 (1.70± 0 .31)mmol/L(P <0 .0 5 ) ,且乳酸含量与细胞凋亡呈正相关(P <0 .0 5 )。　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     

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。     

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.     

高压氧对睾丸扭转/复位后生精细胞凋亡的作用

目的 探讨单侧睾丸扭转/复位后睾丸细胞凋亡状况和高压氧(HBO)的干预作用。方法 32只青春期 Wistar大鼠随机等分为 4组,1组为对照组,2～4组制成单侧睾丸扭转/复位模型(扭转 720°、2 h后复位),3组、4组分别于睾丸复位前和复位后立即予以 60 min HBO治疗。术后48 h获取双侧睾丸,原位缺口末端标记法(TUNEL)观察细胞凋亡,常规染色观察病理学改变。结果 与1组比较,2组扭转侧睾丸生精细胞凋亡增加(P<0.01)、病理损害明显;对侧睾丸生精细胞凋亡增加(P<0.01)。3～4组尤其是4组扭转侧睾丸生精细胞凋亡较2组明显减少(P<0.01)、病理损害减轻;对侧睾丸细胞凋亡较1组无明显变化(P>0.05)。结论 单侧睾丸扭转复位后,双侧睾丸生精细胞凋亡增加,复位前和复位后早期HBO治疗能明显减少细胞凋亡的发生。     

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。     

隐睾动物模型的建立及LHRH对隐睾组织学的影响

为了评价促黄体生成素释放激素（ＬＨＲＨ）对隐睾睾丸的组织学影响。我们用新生家犬制成隐睾动物模型，共分为手术对照组、单侧隐睾组、单侧隐睾加ＬＨＲＨ治疗组及自然隐睾组进行实验研究。研究结果表明，利用外科手术方法在新生犬体内制作的隐睾模型的睾丸组织结构和超微结构与自然隐睾犬睾丸的变化一致，并与人类隐睾的变化相似；单侧隐睾的对侧睾丸未发生交感性改变，小剂量的ＬＨＲＨ可使隐睾睾丸生精细胞数量增加，从而提高其     

总抗氧化能力、一氧化氮、丙二醛对大鼠隐睾生殖细胞凋亡的影响

目的 探讨一氧化氮(NO)、总抗氧化能力(T—AOC)对大鼠隐睾生殖细胞凋亡的影响。方法 采用SD雄性健康大鼠20只，鼠龄22d时复制单侧隐睾模型。采用生物素—dUTP／酶标亲和素测定法检测睾丸生殖细胞凋亡。采用化学比色法测定大鼠组织中丙二醛(MDA)、T—AOC。采用硝酸还原酶法测定睾丸组织中NO含量。结果 术后第7天，与对侧正常睾丸相比，隐睾侧睾丸发生凋亡的生殖细胞数显著增加(P＜0．01)；T—AOC显著下降；No和MDA含量显著上升(P＜0．01)。结论 大鼠隐睾可导致睾丸生殖细胞凋亡增加，且与睾丸组织中No和MDA升高及T—AOC下降密切相关。     

胶质细胞源性神经营养因子对大鼠单侧隐睾组织中干细胞因子及抗氧化酶活性的影响

目的:探讨胶质细胞源性神经营养因子(GDNF)对大鼠隐睾睾丸组织中干细胞因子(SCF)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性及丙二醛(MDA)含量的影响。方法:选取雄性SD大鼠左侧隐睾成功模型48只,体重(200±20)g,完全随机法分为4组,每组12只。A组为对照组,B组为GDNF 7 d组,C组为GDNF14 d组,D组为GDNF 21 d组。每组根据设定处理方式的不同分别进行处理,采集左侧睾丸并行生化指标(SOD、CAT活性及MDA含量)检测;TUNEL检测隐睾组织细胞调亡指数(AI);HE染色观察睾丸组织学形态;实时定量PCR法检测隐睾组织中SCF基因的表达情况,Western印迹检测各组隐睾组织中SCF蛋白表达情况。结果:与GDNF组相比,其中GDNF 14 d组中各项指标有明显变化;生精细胞凋亡明显减少,其中对照组、GDNF 7 d组、GDNF 14 d组、GDNF 21 d组AI分别为13.5±0.64、8.42±0.16、4.45±0.34、7.32±0.09。对照组与GDNF 14 d组相比差异有明显统计学意义(P<0.01),与对照组相比,各GDNF组生精细胞凋亡减少,SOD活性明显上升,MDA含量下降明显,SCF表达较对照组有明显增高,差异有明显统计学意义(P<0.01)。结论:GDNF对大鼠单侧隐睾组织生精功能具有保护作用,增强抗氧化酶系统的抗氧化能力,提高SCF表达含量,从而改善睾丸生育能力,其以GDNF 14 d时作用最明显。     

单侧隐睾睾丸组织中胰岛素样因子3基因表达

目的 研究单侧隐睾对睾丸组织胰岛素样因子3（Insl3）表达的影响。方法 通过建立隐睾大鼠动物模型获得A组单侧隐睾大鼠12只和B组双侧隐睾大鼠10只，C组空白对照正常大鼠10只。在大鼠8周龄时取睾丸组织用Tarazol法提取总RNA，Northern Blot方法检测各组睾丸组织中Insl3基因表达水平；同时提取各组睾丸组织蛋白，Westernblot检测Insl3蛋白多肽表达。结果 Insl 3 mRNA在单侧隐睾侧睾丸组织中表达量低于C组（P〈0．05），单侧隐睾大鼠对侧睾丸表达量低于C组（P〈0.05）；InSl3基因表达的蛋白多肽在隐睾侧睾丸组织中表达量较低（P〈0．05），对侧睾丸组织蛋白表达较C组低（P〈0．05），但较隐睾侧睾丸和B组明显增多（P〈0．05）。结论 单侧隐睾生精功能的损害可能是影响了隐睾侧睾丸Insl3基因表达，同时还抑制对侧睾丸的表达。     

环孢素对大鼠睾丸缺血-再灌注损伤后生精细胞凋亡的影响

目的探讨环孢素对大鼠单侧睾丸缺血-再灌注损伤后生精细胞凋亡的影响,研究其保护性作用。方法雄性SD大鼠24只,随机分成假手术组、扭转组和环孢素组,每组8只,建立720°2h单侧睾丸扭转动物模型。扭转组和环孢素组于复位前15min分别腹腔注射生理盐水和环孢素,术后24h留取手术侧睾丸。应用流式细胞术检测各组患侧睾丸组织生精细胞凋亡;Quantitative Real-time PCR技术对Fas、FasL和Bax mRNA进行定量分析;Western-blot技术检测细胞色素C含量。结果与假手术组相比,扭转组患侧睾丸组织早期凋亡生精细胞百分比明显增多,Fas、FasL和Bax mRNA表达上调,同时胞质中细胞色素C含量显著升高,其差异均有显著性(P均<0.05)。环孢素干预能显著减轻上述变化,患侧睾丸组织正常细胞群百分比升高,Fas、FasL和Bax mRNA表达下调,同时胞质中细胞色素C含量显著降低,其差异均有显著性(P均<0.05)。结论环孢素可能参与调控生精细胞凋亡的分子途径,对睾丸扭转术后的生精功能具有明显的保护性效果。     

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.     

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     

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.     

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。     

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。     

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.     

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.