Pre-ischemic administration of nitric oxide synthase inhibitors reduced germ cell apoptosis after spermatic vessel ligation in the rat testis

INTRODUCTION: The Fowler-Stephens maneuver, a mode of spermatic vessel ligation, is a method of choice in the management of high testes. The aim of the present study is to investigate the effects of pre-ischemic administration of N(G)-nitro-L-arginine methyl ester (L-NAME), a nitric oxide (NO) synthase (NOS) inhibitor, in preventing testicular ischemic damage and germ cell-specific apoptosis in rats. MATERIALS AND METHODS: 30 min before ligation of the spermatic vessels, L-NAME was administered intraperitoneally to a group of rats and saline was given to another group of rats (controls). Biochemical assessments of NO and germ cell-specific apoptosis were performed. RESULTS: Testicular NO levels in the L-NAME group showed significant decreases in the ipsilateral (p = 0.004) and contralateral (p = 0.015) testes in relation to those of the control group. The apoptotic indices were found in 2.3% of the L-NAME group and 3.1% of the control group. CONCLUSION: Pre-ischemic administrations of the NOS inhibitor, L-NAME, effectively decreased NO production and to some degree caused a reduction in germ cell apoptosis in the rat testes after spermatic vessel ligation. Further studies are mandatory to confirm our preliminary results and to address the potential introduction of NOS inhibitors into clinical practice.     

Collateral circulation after spermatic vessel ligation for abdominal testis and its impact on staged laparoscopically assisted orchiopexy

PURPOSE: Staged laparoscopically assisted orchiopexy for abdominal testis entails initial spermatic vessels ligation followed by mobilization of the testis, preserving the vas and its vessels as a sole source of testicular blood supply. This mobilization includes all peritoneal attachments of the testis, including the gubernaculum, which may carry collateral circulation to the testis. This study considers the anatomy of the gubernaculum and the collateral circulation after spermatic vessels ligation and its possible effects on the viability of the testis. MATERIALS AND METHODS: The anatomy of the gubernaculum and the effect of spermatic vessels ligation on the collateral circulation were studied in 90 boys with 100 abdominal testes with a short pedicle. Patients with vanishing testis or those not needing ligation of the spermatic vessels were excluded from the study. The anatomy and the vascularity of the testis, gubernaculum, and vas were studied at initial laparoscopy and 6 weeks later after spermatic vessel ligation. Based on these findings, the technique for laparoscopically assisted orchiopexy were modified, preserving the gubernaculum whenever possible in cases having prominent collaterals. RESULTS: Based on the attachment and blood vessel configuration of the gubernaculum, the patients were divided into two groups: in group 1 (open internal ring), there were 46 testes in which the gubernaculum passed through an open internal ring having an inguinal attachment. In group 2 (closed internal ring), there were 54 testes with a soft gubernaculum attached to a closed internal ring without inguinal attachment. In group 1 the gubernaculum was short and tough in 32 of 46 testes, with no visible blood vessels in all cases. In group 2 the gubernaculum was long and soft in 43 of 54 testes and showing evident blood supply before clipping of the spermatic vessels in 30 testes. Subsequent laparoscopy done after 6 weeks showed prominent collateral circulation around the gubernaculum in 26 testes and around the vas in 20 testes in the group 1 patients, and around the vas in 20 and the gubernaculum in 34 testes in group 2. Preservation of the gubernaculum was possible in 43/54 (80%) of group 2 patients and in 14/46 (30%) of group 1 patients. CONCLUSION: Routine cutting of the gubernaculum is not necessary for proper mobilization of the abdominal testis: collateral circulation varies from patient to patient, and once the gubernaculum shows prominent blood supply, its preservation is mandatory. The decision to cut the gubernaculum should be taken while the performing initial spermatic vessel ligation and not during the second stage to avoid compromise of the settled collateral circulation.     

腹腔镜保留睾丸动脉精索静脉高位结扎术对睾丸生精功能的影响

目的 探讨腹腔镜下精索内静脉高位结扎同时保留睾丸动脉治疗精索静脉曲张的应用价值.方法 选取156例精索静脉曲张患者,随机分成A、B两组:A组采用腹腔镜下保留睾丸动脉精索静脉高位结扎术,B组采用传统的Palomo术式.比较两组手术前后精液质量参数变化、术后并发症的发生率.结果 A组的手术时间为(32.24士8.35)min,明显短于B组(36.45士9.14)min (P＜0.05),术后阴囊水肿(2.67％)、附睾炎(4％)、睾丸萎缩率(2.67％)明显低于B组(P＜0.05),住院时间、术后复发率比较无统计学意义(P＞0.05);术后精子计数[(43.00±4.3)×106]、精子活率[(65.00±4.55)％]明显高于B组[(37.20士5.45)×106]、[(53.34±6.73)％](P＜0.05),精子密度、精子活力分级比较差异无显著性(P＞0.05).结论 保留睾丸动脉腹腔镜精索内静脉高位结扎术能够明显提高术后精液参数质量,降低阴囊水肿、附睾炎、睾丸萎缩的发生率.     

Effects of testicular fixation on oxidative stress-related parameters in prepubertal rat testis

The goal of orchiopexy is to provide adequate scrotal fixation to prevent recurrent torsion of the testis and the spermatic cord or ascent of the testis, to be achieved with minimal trauma to the testis. Forty-two prepubertal male Sprague-Dawley rats were equally distributed into 6 groups (1 SHAM + 5 Operation). In operational groups, two lateral sutures were placed near the upper and lower pole of the testis passing through the tunica albuginea with 4-zero polyglactin suture in the transverse axis of the testis. At the 1st-3rd-7th-15th and 30th postoperative days, bilateral orchiectomy was performed, then malondialdehyde (MDA), nitrite, superoxide dismutase (SOD) and glutathion peroxidase (GPx) values were measured in testicular tissue homogenates. MDA and nitrite levels increased, while GPx and SOD levels diminished, statistically significant in both ipsilateral and contralateral testicular tissue homogenates. Prepubertal testicular fixation causes overproduction of the lipid peroxidation end product and free radicals while heavy consuming anti-oxidant reserve systems in both ipsilateral and contralateral testicular tissue. These results clearly reveal the extent of the testicular deterioration due to suture fixation technique.     

Hemorrhagic infarctions of testis due to intimal fibroplasia of spermatic artery

Two cases of hemorrhagic infarctions of the testis are presented. The infarctions were due to intimal fibroplasia of the branches of the spermatic artery; to our knowledge, a condition described only in the kidney. The differences between intimal fibroplasia and the lesions of polyarteritis nodosa and thromboangiitis obliterans are discussed.     

Long-term effect of pentoxifylline and NG-nitro-L-arginine methyl ester on testicular function in spermatic artery ligation

OBJECTIVE: To investigate long-term testicular function in pentoxifylline- and NG-nitro-L-arginine methyl ester (L-NAME)-treated rats prior to spermatic artery ligation by assessing serum inhibin B concentration, a reliable endocrine marker of spermatogenesis. MATERIAL AND METHODS: Forty prepubertal rats were randomly divided into four equal groups. Right orchiectomy was performed in all rats. Intraperitoneal pentoxifylline and L-NAME were administered to Groups 1 and 2, respectively. Rats in Group 3 did not receive any medication. The spermatic arteries of the left testes were ligated in all groups with the exception of the sham controls (Group 4). One month postoperatively, a histopathologic evaluation was performed and serum inhibin B concentrations were assessed in all groups. RESULTS: There were no statistically significant differences in testis size or serum inhibin B concentration between Groups 1, 2 and 3. However, the testes were significantly smaller (p(1)=0.01, p(2)=0.01, p(3)=0.01) and serum inhibin B levels were significantly decreased (p(1)=0.01, p(2)=0.01, p(3)=0.01) when Groups 1-3 were compared with Group 4. Histopathological evaluations revealed necrosis and calcification in all specimens with the exception of the sham-operated group. CONCLUSIONS: The long-term outcomes of spermatic artery ligation were poor and testicular atrophy developed in rat testes in all study groups. Testicular atrophy could not be reversed by preoperative administration of pentoxifylline and L-NAME.     

Effect of experimental varicocele in rats on testicular oxidative stress status

The present experiments were undertaken to determine the levels of MDA, SOD and catalase in the testis of adolescent rats with experimental left varicoceles. Male Wistar rats, 7 weeks old and weighing 160-170 g, were randomly allocated into three groups. The first group of rats underwent partial ligation of the left renal vein (n = 15). The second group of rats underwent a sham operation (n = 7) and the third group acted as controls (n = 7). Animals were sacrificed 6 weeks after surgery and dilatation of the internal spermatic veins was observed. Levels of MDA, SOD and catalase activity were measured in testis. The experimental left varicocele group showed severe testicular changes compared to other groups. The mean MDA (SEM) levels in right and left testicular tissues of varicocele bearing rats, sham-operated rats, and control rats were 0.48 +/- 0.24 and 0.31 +/- 0.11, 0.22 +/- 0.02 and 0.35 +/- 0.12, 0.62 +/- 0.29 and 0.13 +/- 0.05, respectively (P > 0.05). The mean SOD (SEM) levels in right and left testicular tissues of varicocele bearing rats, sham-operated rats, and control rats were 7,790 +/- 606 and 6,974 +/- 574, 7,475 +/- 1,517 and 7020 +/- 1,106, 8,727 +/- 1,188 and 9,019 +/- 1,129, respectively (P > 0.05). The mean catalase (SEM) levels in right and left testicular tissues of varicocele bearing rats,sham-operated rats, and control rats were 75.77 +/- 11.5 and 53.82 +/- 10.1, 91.94 +/- 14 and 94.90 +/- 32, 65.40 +/- 5.7 and 90.93 +/- 16.4, respectively (P > 0.05). Our results suggest that oxidative status, which reflects a relative balance between reactive oxygen species (ROS) generated and ROS scavenged, may not be responsible for the testicular dysfunction associated with experimentally induced varicocele during adolescence in rats.     

Postoperative hydrocele testis as a complication of high ligation of internal spermatic vein

We reviewed 785 patients with varicocele testis who underwent high ligation of internal spermatic vein at Kasukabe City Hospital between 1969 and 1985, and found only 3 patients suffering from postoperative hydrocele testis. Analysis of the hydrocele content in one case revealed a high protein level, which suggested that the etiology of the hydrocele after high ligation is of lymphatic origin, so that it is important to preserve the lymphatics of spermatic cord in order to prevent postoperative hydrocele.     

Effects of spermatic vascular division for correction of the high undescended testis on testicular function

Orchiopexy with division of the spermatic artery and veins is a commonly used technique for correcting the high undescended testis, although the longterm results have not been clearly defined. The left spermatic artery and veins of 22 adult Wistar albino rats were divided while preserving the vessels associated with the vas and cremaster muscle (DT). A sham operation was performed on the left testicle of six additional rats (ST). At 3 weeks postoperatively, both testes from all rats were removed. All testes were viable and bled when incised, although bleeding was considerably reduced in testes with DT. Mean testicular weights after DT were 1,061 +/- 423 mg compared with 1,634 +/- 125 mg for ST rats (p less than 0.02) and 1,508 +/- 119 mg for contralateral testes. The mean tubular diameter after DT was 220 +/- 37 mu compared with 303.1 +/- 10.7 mu for ST testes (p less than 0.02). The testicular biopsy score based upon the morphology of the spermatic tubules was 4.46 +/- 3.32 for DT testes and 8.65 +/- 0.23 for ST testes (p less than 0.02) compared with 8.38 +/- 0.18 for contralateral testes and an absolute normal value of 10. No morphologic abnormalities were observed in the contralateral unoperated testes from any of the rats. The contralateral testes in 12 additional rats were removed before DT. The mean testosterone values in these rats with one testicle was 1.43 +/- 0.75 ng/mL. Three weeks after DT, testosterone values were 0.19 +/- 0.31 ng/mL (p less than 0.01). It is concluded that division of the main spermatic artery and vein in rats produces testicular atrophy with spermatogenic arrest and interstitial cell dysfunction. Although collateral blood flow to the testis may be demonstrated, tissue perfusion is inadequate for normal spermatogenesis and endocrine function.     

Laparoscopic spermatic vessel ligation in the treatment of varicocele: report of 129 procedures

With the improvement of laparoscopic techniques in the hands of urologists, a minimally invasive therapy can be offered to patients. We report retrospectively our experience in the laparoscopic treatment of varicoceles. A total of 93 patients with 129 varicoceles were treated by a laparoscopic approach, the mean age was 29 years (12-47), with 57 left varicoceles and 36 bilateral varicoceles. Indications for treatment were infertility in 51 patients and pain in 42 cases. The mean duration of the operation was 40 min (25-85) for bilateral varicoceles, and 24 min (15-65) for left varicoceles. All our patients were operated the day of their admission and 52 patients left the hospital 8 h after the intervention. We report 2 incidents with conversion to open surgery in one case. The pregnancy rate after surgery in patients with infertility was 21%, the recurrence rate was 4%. Laparoscopic spermatic vein ligation in the treatment of varicocele is a minimally invasive procedure, simple, fast and is not a contraindication for other procedures in case of recurrence.     

Effect of testicular torsion on the contralateral testis

It has been previously shown that unilateral testicular torsion can cause disruptive anatomic changes in the contralateral testis in rats [1]. In this experimental study plasma and urine prostaglandin E2 levels were studied correlatively with testicular histopathology in acute testicular torsion cases. As a result of this study, necrobiotic morphologic alterations causing testis necrosis and significant increase in plasma prostaglandin E2 levels were observed. Contralateral testicular histology was analyzed in all dogs. None of them showed abnormal tubular architecture.     

Paeonol protects against testicular ischaemia-reperfusion injury in rats through inhibition of oxidative stress and inflammation

Ischaemia-reperfusion (IR) is the most common form of testicular injury that results in oxidative damage and inflammation ending by subinfertility. Paeonol, a natural phenolic compound, exhibits antioxidant and anti-inflammatory effects. Thus, the present study investigated the role of paeonol in rat testicular IR injury. Thirty adult Wistar rats were randomly divided into five groups; sham, sham treated with paeonol, IR injury, and IR pre-treated with paeonol at low and high doses. Serum testosterone and testicular levels of malondialdehyde and reduced glutathione (GSH) besides superoxide dismutase (SOD) activity were determined. Gene quantifications for tumour necrosis factor-α (TNF-α), hypoxia-inducible factor-1α (HIF-1α) and heat shock protein 70 (HSP70) were also assessed. Histopathological pictures and the immunohistochemical expression of testicular nuclear factor erythroid 2-related factor 2 (Nrf2), interleukin-1β (IL-1β) and interleukin-6 (IL-6) were shown. Pre-treatment with paeonol prevented the drop in serum testosterone, alongside with improvement of testicular malondialdehyde and GSH levels plus SOD activity. Paeonol regained the normal spermatogenesis with prevention of IR-induced increase in TNF-α, HIF-1α and HSP70 gene expression besides IL-1β and IL-6 immunostaining and reduction in Nrf2 protein expression. Paeonol exerted a dose-dependent beneficial effect on testicular IR injury. This effect was achieved by its antioxidant and anti-inflammatory effects.     

Iron-induced injury of rat testis

Summary. Male Wistar rats were intraperitoneally injected twice with 0.4 mmol kg⁻¹ FeSO₄. One, 2 and 4 days after the second Fe injection, Fe and malondialdehyde (MDA) content in testis was measured, the morphology studied by light and electron microscopy and the number of spermatids counted. After Fe injection, Fe and MDA content had increased in parallel. Light microscopic inspection on days 1 and 2 after Fe injection revealed numerous necroses in the different cell types of the germ epithelium. Four days after Fe injection, fewer alterations were found. Electron microscopic investigations revealed that some spermatids contained up to three nuclei and at least three axonemes. In some sperm tails up to 11 axonemes were found. In some midpieces two or three complexes of axonemes, outer dense fibres and mitochondria were observed. In other midpieces axonemes were absent and replaced by granular and filamentous material. The number of spermatids was reduced 4 days after Fe treatment. The increase in the number of axonemes was similar to that seen in Mg and Zn deficiency, indicating that the increase in Fe content and oxygen free radicals is the major reason for the biochemical and morphological alterations in Mg and Zn deficiency.     

单侧睾丸扭转术后健侧睾丸功能的研究

目的研究睾丸扭转患者术前和术后不同时间性激素、抑制素B及精液参数的动态变化，探讨健侧睾丸的损伤与修复代偿功能。方法10例睾丸扭转患者（16～45岁），睾丸扭转病程3～6d，平均4．7d；左侧9例，右侧1例，术中探查示患侧睾丸坏死，全部行患侧睾丸切除。全自动化学发光法检测血清中FSH、LH、T的值；ELISA法定量检测患者血清中抑制素INHB的浓度；WLJY-9000分析精液参数。结果INHB水平在术后3周水平最低，12周时恢复正常，术前与术后3周、6周之间有显著差异；术后26周血LH和INHB水平相对术后6周时显著升高；精液常规检测示，术后3周时精液中精子数、精子活率、精子活力较正常标准明显降低，12周时有好转，26周时基本恢复正常：FSH与INHB呈显著负相关P〈0．05，R=-0．900，INHB与精子数量及精子存活率呈显著正相关P〈0．01，R=1．000。结论睾丸扭转者患侧睾丸切除后，健侧睾丸有一个损伤和修复的过程；血清INHB和T变化是睾丸功能的敏感指标，能较准确地反映睾丸损伤和修复程度。     

参精固本丸对大鼠睾丸氧化应激损伤后生精功能的修复

目的:探讨补肾活血中药参精固本丸对氯化镉造模的氧化应激损伤大鼠睾丸、附睾、精子的抗氧化及生精功能修复作用。方法:将SPF级Wistar大鼠随机分为6组,每组12只,分别为正常对照组、模型对照组、五子衍宗丸组、参精固本丸高剂量组、参精固本丸中剂量组、参精固本丸低剂量组。除正常对照组外,各组腹腔注射氯化镉1 mg/kg,24 h后各组分别灌胃给药,每天1次,共给药56 d。模型对照组和正常对照组予等体积生理盐水灌胃,参精固本丸高、中、低剂量组分别予以8倍等效剂量(11.2 g/kg)、4倍等效剂量(5.6 g/kg)、2倍等效剂量(2.8 g/kg)参精固本丸溶液灌胃,五子衍宗丸组予以4倍等效剂量(4.5 g/kg)五子衍宗丸溶液灌胃。56 d后,处死实验动物,分别称取大鼠精囊、附睾、睾丸的质量并计算相应脏器系数,大鼠睾丸、附睾、精囊行病理组织学检测。留取附睾内精液,测定大鼠附睾精子浓度、活力、正常形态精子百分率。同时测定大鼠睾丸组织谷胱甘肽过氧化物酶(GSH-PX)、超氧化物歧化酶(SOD)、丙二醛(MDA)水平,血清睾酮(T)水平。结果:①参精固本丸高、中、低剂量组睾丸系数分别为(0.403±0.090)、(0.357±0.150)、(0.348±0.140)g/100 g,精囊系数分别为(0.347±0.115)、(0.336±0.090)、(0.320±0.065)g/100 g,高剂量组附睾系数为(0.156±0.030)g/100 g,均显著高于模型对照组[(0.237±0.098)、(0.241±0.118)、(0.099±0.088)g/100 g](P0.05或P0.01);②高、中、低剂量组精子浓度分别为(58.1±32.2)、(36.0±36.2)、(31.9±32.7)×10~6/ml,显著高于模型对照组[(10.5±17.7)×10~6/ml](P0.05或P0.01);高、中剂量组精子总活力分别为(26.5±15.5)%、(18.9±8.2)%,高、中剂量组正常形态精子百分率分别为(85.3±23.3)%、(65.8±28.1)%,均显著高于模型对照组[(9.5±13.0)%、(36.2±40.2)%](P0.05或P0.01);③参精固本丸高、中剂量组睾丸的GSH-PX水平分别为(5.70±1.73)、(5.42±2.35) U/mg prot,SOD水平分别为(52.7±14.6)、(51.3±14.7) U/mg prot,均显著高于模型对照组[(3.62±2.22)、(41.3±8.8)U/mgprot](P0.05或P0.01);高、中、低剂量组睾丸MDA水平分别为(0.41±0.29)、(0.44±0.19)、(0.47±0.20) nmol/mgprot,显著低于模型对照组[(0.69±0.28) nmol/mgprot](P0.05或P0.01);高、中、低剂量组大鼠血清T水平分别为(3.62±0.96)、(3.48±1.33)、(3.24±0.83)nmol/L,显著高于模型对照组[(2.56±0.75) nmol/L](P0.05或P0.01);④模型对照组全部生精小管均发生凝固性坏死并钙化,生精细胞及支持细胞消失,精囊管腔内分泌物减少,上皮乳头萎缩;附睾萎缩,管腔缩小,管腔内未见精子。与模型对照组相比,随着剂量增加,参精固本丸组具有生精功能的生精小管面积逐渐增加,精囊上皮恢复为高柱状,附睾管管腔逐渐增大,管腔内精子逐渐增多,疗效与剂量间有依从关系,参精固本丸高、中剂量组疗效明显好于五子衍宗丸组。结论:参精固本丸可以拮抗氧化应激损伤,提高实验大鼠睾丸抗氧化物酶、血清T水平、精液质量,修复睾丸、附睾、精囊组织病理损伤,改善生精功能。     

Effect of orchiectomy on sperm functional aspects and semen oxidative stress in men with testicular tumours

To verify if quality of spermatozoa from men with testicular germ cell tumours is better before or after orchiectomy. This prospective study was carried out including 24 patients with testicular germ cell tumours, who provide one semen sample before they were submitted to unilateral orchiectomy and one other semen sample 30 days after the surgery. After collection by masturbation and liquefaction, an aliquot of the semen sample was used for semen analysis and another was used to evaluate sperm mitochondrial activity, DNA fragmentation and acrosome integrity. Seminal plasma was used to evaluate lipid peroxidation levels. Pre‐orchiectomy sample and post‐orchiectomy sample were compared using a paired Student's t test (normal distribution) or a paired Wilcoxon test, when appropriate (p ? 0.05). No significant difference was observed in semen analysis. A significant decrease in DNA fragmentation and lipid peroxidation and an increase in mitochondrial activity were observed after orchiectomy. Based on our findings, the semen quality from men with testicular germ cell tumours is better after orchiectomy.