Proxeed plus salvage rat testis from ischemia- reperfused injury by enhancing antioxidant's activities and inhibition of iNOS expression.

Testicular torsion is an acute urological emergency condition that occurs due to obstruction of blood flow to the testicles which may result in ischemia and loss of testicular functions. This study examined the protective effects of Proxeed Plus (PP), a dietary supplement on testicular ischemia/reperfusion (I/R) injured rats using oxidative stress markers, hormonal levels, apoptotic parameters, histological and immunohistochemistry analysis at 4 h and after 7 days of reperfusion. The protective treatment of the I/R injured rats with PP at 1000 and 5000 mg/kg body weight (bw) resulted in significant increases in the serum and tissue antioxidative defense capacities (superoxide dismutase, reduced glutathione, catalase, glutathione-s-transferase, and glutathione peroxidase), sex hormones (luteinizing hormone, follicle-stimulating hormone, and testosterone), also reduce pro-oxidative markers (malondialdehyde and hydrogen peroxide), serum iNOS and apoptotic parameters (Caspase -3 and Caspase -9) in comparison to the results detected in the I/R untreated rats. It was also observed that PP ameliorated histological changes of I/R injured rats; increased spermatogenetic activity, seminiferous tubular diameter, Leydig cell mass, and reduced expressions of testicular inducible nitric oxide synthase (iNOS). Therefore, the therapeutic use of Proxeed Plus could be considered a promising approach in averting testicular damage against I/R injury.

The efficiency of Poly(ADP-Ribose) Polymerase (PARP) cleavage on detection of apoptosis in an experimental model of testicular torsion.

The aim of this study was to evaluate the histopathological and apoptotic changes occurring in the rat ipsilateral and contralateral testes, after experimental spermatic cord torsion, and to explore and the role of poly(ADP-ribose) polymerase (PARP) cleavage in testicular torsion-detorsion injury. A total of 37 Wistar albino rats were subjected to 720° unilateral spermatic cord torsion for 1, 2 and 4 h, followed by 4-h reperfusion, or else to a sham operation (control group). Histology of the testicle was evaluated using haematoxylin-eosin (H&E) staining and Johnsen's scoring system. Germ cell apoptosis was evaluated via active caspase-3 immunostaining, and PARP expression levels were evaluated via Western blotting. The mean Johnsen's tubular biopsy scores (JTBS) of the ipsilateral testicles were lower for all torsion groups than for the controls (P < 0.05), but the JTBS of the contralateral testicles were only lower in the 4-h torsion group (P < 0.05). The mean apoptosis score (AS) of the ipsilateral and contralateral testicles was significantly higher in the torsion groups than in the sham group. AS increased correlatively with torsion time, in both testicles. The effect of testicular torsion on PARP cleavage was time dependent, with the highest effect observed after 4 h of testicular torsion (P < 0.05). Testicular torsion caused time-dependent histological changes, apoptosis and increases in PARP cleavage. Our results suggest that testicular torsion-detorsion injury caused cell damage and germ cell apoptosis that apparently involved cleavage of PARP. Increased PARP cleavage could, in turn, lead to enhanced apoptosis.

Coenzyme Q10 treatment reduces lipid peroxidation, inducible and endothelial nitric oxide synthases, and germ cell-specific apoptosis in a rat model of testicular ischemia/reperfusion injury.

In this experimental study, we assessed the preventive effects of coenzyme Q(10) (CoQ(10)) in a rat model of ischemia/reperfusion injury. The results of this study show that CoQ(10) administration before the reperfusion period of testicular torsion provides a significant decrease in testicular lipid peroxidation products and expressions of inducible nitric oxide synthase, endothelial nitric oxide synthase, and germ cell-specific apoptosis.

Ginkgo biloba (EGB 761) affects apoptosis and nitric-oxide synthases in testicular torsion: an experimental study.

INTRODUCTION: We investigated the effect of ginkgo biloba on germ cell apoptosis and also on expressions of endothelial (eNOS) and inducible (iNOS) nitric oxide synthases after testicular torsion. MATERIALS AND METHODS: Thirty-two Wistar albino rats were randomly assigned into four groups. Torsion/detorsion (T/D) was performed on the rats in group 1, group 2 received ginkgo biloba for a month before T/D, group 3 received only gingko biloba for a month, and group 4 was the sham group. Left testicular torsion was created in group 1 and group 2, and the testes were untwisted and replaced in the scrotum for reperfusion. No procedure was applied to group 3, and after 1 month, testes were removed in all groups. RESULTS: Mean apoptotic cell, eNOS, and iNOS were increased in group 1. Group 2 showed significantly decreased apoptotic cells, eNOS, and iNOS in testes compared to group 1 (P < 0.05). The rats in group 3 had significantly decreased apoptotic cell, eNOS, and iNOS values, like the sham group (P < 0.05), and this group provided basal values. CONCLUSIONS: Ginkgo biloba, as a free radical scavenger, seems to have a protective role against apoptosis in testicular ischemia reperfusion injury.

Hemin induced heme oxygenase-1 over expression involves nuclear factor-E2 related factor-2, nuclear factor-kappaB and extracellular regulated kinase: an experimental study in a testicular torsion-detorsion rodent model.

PURPOSE: Nrf2 (nuclear factor-E2 related factor-2), nuclear factor-kappaB and mitogen-activated protein kinase, including extracellular regulated kinase, c-jun N-terminal kinase and p38 mitogen-activated protein kinase, are crucial for signal transduction. We previously reported that heme oxygenase-1 over expression induced by the heme oxygenase-1 inducer hemin protected testes from torsion-detorsion injury. In this study we elucidated which of these enzymes is involved in hemin induced heme oxygenase-1 over expression in testicular tissues after torsion-detorsion injury. MATERIALS AND METHODS: Adult male Sprague-Dawley rats (National Science Council, Taipei, Taiwan, Republic of China) were allocated to undergo testicular torsion-detorsion immediately followed by injection of normal saline, hemin or hemin plus tin protoporphyrin, a heme oxygenase-1 inhibitor. Another set of rats that underwent sham operation were used as controls. Testes were harvested 0 and 30 minutes after detorsion and enzyme expression was analyzed. RESULTS: Hemin alone caused no significant effects on the expression of Nrf2, nuclear factor-kappaB, extracellular regulated kinase and c-jun N-terminal kinase. Similarly testicular torsion-detorsion injury caused no significant effects on Nrf2 expression. In the presence of torsion-detorsion injury hemin significantly up-regulated Nrf2 expression. Moreover, this effect was not affected by tin protoporphyrin. Unlike Nrf2, the expression of nuclear factor-kappaB, extracellular regulated kinase and c-jun N-terminal kinase was significantly up-regulated by testicular torsion-detorsion. Hemin significantly attenuated the testicular torsion-detorsion induced up-regulation of nuclear factor-kappaB and extracellular regulated kinase but not c-jun N-terminal kinase. The effects of hemin on nuclear factor-kappaB and extracellular regulated kinase were significantly reversed by tin protoporphyrin. However, the expression of p38 mitogen-activated protein kinase in rodent testes was not affected by these interventions. CONCLUSIONS: Hemin induced heme oxygenase-1 over expression in rodent testes after torsion-detorsion injury involves Nrf2, nuclear factor-kappaB and extracellular regulated kinase.

The protective role of erdosteine on testicular tissue after testicular torsion and detorsion.

Testicular torsion and detorsion are important clinical problems for infertile man and oxidative stress may have a role in this clinical situation. The aim of this study was to investigate the protective role of erdosteine, an antioxidant, on unilateral testicular reperfusion injury in rats. The rats were divided into four groups including seven rats in each group: control, torsion, torsion/detorsion and torsion/detorsion+erdosteine. Rats, except the sham operation group, were subjected to left unilateral torsion (720( composite function) rotation in the clockwise direction) without including the epididymis. The experiments were finished after sham operation time for control, 120 min torsion for torsion group and 120 min torsion and 240 min detorsion for torsion/detorsion groups. Bilateral orchiectomy was performed for all groups of rats. The ipsilateral and controlateral testis were divided into two pieces to analyse biochemical parameters and to investigate the light microscopic view. Malondialdehyde level of ipsilateral testis was increased in torsion and torsion/detorsion groups in comparison with the other groups (p < 0.05). Erdosteine treatment ameliorated lipid peroxidation after torsion/detorsion in ipsilateral testis (p < 0.05). Also, xanthine oxidase activity of ipsilateral testis was increased in torsion/detorsion group in comparison with the others (p < 0.05). Nitric oxide (NO) level of ipsilateral testis was higher in all experimental groups than sham operated control group (p < 0.05). Also, NO level of torsion group was increased in comparison with detorsion groups (p < 0.05). Erdosteine treatment caused increased glutathione peroxidase activity in comparison with torsion and torsion/detorsion groups and catalase activity in comparison with the other groups in ipsilateral testis (p < 0.05). Superoxide dismutase activity of ipsilateral testis was higher in torsion/detorsion and torsion/detorsion+erdosteine groups than control and torsion groups (p < 0.05). The biochemical parameters were not affected in controlateral testis in all groups. Torsion, torsion/detorsion and torsion/detorsion+erdosteine groups showed ipsilateral testicular damage in the histological examination, but the specimens from torsion/detorsion had a significantly greater histological injury than those from the other groups (p < 0.05). Control rats showed normal seminiferous tubule morphology. Rats in torsion group had slight-to-moderate disruption of the seminiferous epithelium. Rats in torsion/detorsion group displayed moderate-to-severe disruption of the seminiferous epithelium. In all animals from torsion/detorsion+erdosteine group, the testicular tissues were affected with slight-to-moderate degenerative changes of the seminiferous epithelium. Administration of erdosteine resulted in a significantly reduced histological damage associated with torsion of the spermatic cord compared with torsion/detorsion. In all groups, the contralateral testes were histologically normal. In conclusion, the results clearly displayed that erdosteine treatment may have a protective role on testicular torsion/detorsion injury.

The effect of zinc aspartate pretreatment on ischemia-reperfusion injury and early changes of blood and tissue antioxidant enzyme activities after unilateral testicular torsion-detorsion.

BACKGROUND/PURPOSE: The aim of this study is to investigate the effect of zinc aspartate (ZA) pretreatment on ischemia-reperfusion (I/R) injury and blood and tissue antioxidant enzyme activity early after unilateral testicular torsion-detorsion (T/D). METHODS: Forty prepubertal male Sprague-Dawley rats (weight 160 to 220 g) were divided into 4 groups each containing 10 rats. Surgery was conducted under intraperitoneal 1-shot ketamine (50 mg/kg) anesthesia. The scrotum was entered through a midline incision. Rotating the left testis 720 degrees in a clockwise direction was the model of the testicular torsion. Group 1 was for the basal values. Group 2 had 4 hours T/D. Group 3 also had 4 hours T/D and pretreated with 50 mg/kg intraperitoneal ZA injection half an hour before detorsion. Group 4 was designed as a sham group. In the Group 2 and Group 3, the tunica vaginalis was opened, and left testicles were rotated clockwise 720 degrees and maintained in this torsion position by fixing with a silk suture to the scrotal wall. The scrotum was closed and 4 hours later reentered for testicular detorsion. After spermatic cord detorsion, the scrotum was closed. At the end of 4 hours detorsion period, bilateral orchiectomies were performed, and 5-mL intracardiac blood samples were taken. Blood and tissue superoxide dismutase (SOD), catalase (CAT) activities, and malondialdehyde (MDA) levels were measured, and histopathologic examination was performed. RESULTS: Group 2 and group 3 had decreased blood SOD and CAT activities and elevated MDA levels indicating I/R injury. The 2 groups were also different from each other for these parameters reflecting the beneficial effect of ZA pretreatment (P <.05). The decreased ipsilateral tissue SOD and CAT activities in group 2 were different from the other groups including group 3 (P <.05). Ipsilateral tissue MDA levels of both group 2 and group 3 were elevated. Group 2 had higher values than group 3 (P <.05). In addition, specimens from group 2 had a significantly greater histologic injury than group 3 (P <.05). These findings were also supporting the beneficial effect of ZA pretreatment. All measurements of contralateral tests were similar to the basal values for all groups (P >.05). CONCLUSIONS: ZA pretreatment reduces I/R injury by its antioxidant effects after unilateral testicular T/D and affects the antioxidant enzyme systems.

Activation of endothelial nitric oxide synthase in contralateral testis during unilateral testicular torsion in rats.

There are controversies about the injury of the contralateral testis during unilateral testicular torsion (UTT). An autonomic reflex arc between bilateral testes has been proposed. The authors focused on the involvement of nitric oxide (NO) in the contralateral testis during UTT. Eight-week-old male Wistar rats underwent unilateral torsion (1 h)-detorsion (up to 24 h). NO synthase (NOS) activity was detected as NADPH-diaphorase activity after fixation by paraformaldehyde. N-nitro-L-Arginine methyl ester (L-NAME, 20 mg/kg) was injected intravenously to the other group of rats. To evaluate the testicular injury, proteolysis of alpha-fodrin production was detected by Western blotting. Apoptosis of the germ cells was evaluated by TUNEL. Long-term effect on spermatogenesis was evaluated by flow cytometry at 60 days after UTT. Transient activation of NOS was detected following the proteolysis of alpha-fodrin in the contralateral testis. L-NAME inhibited these alterations. NADPH-diaphorase activity and eNOS immunoreactivity were co-localized in the endothelial cells. These reactions were not observed in other organs. There was neither enhanced apoptosis nor deteriorated spermatogenesis in the contralateral testis during and 60 days after UTT. In the contralateral testis, eNOS-derived NO regulates the vasomotor function against unilateral testicular torsion, whereas it acts slightly cytotoxic. These results suggest the possible involvement of a testis-specific neurovasomotor reflex between the bilateral testes.

The effect of poly (adenosine diphosphate-ribose) polymerase inhibitors on biochemical changes in testicular ischemia-reperfusion injury.

PURPOSE: Poly (adenosine diphosphate [ADP]-ribose) polymerase inhibitors have been used successfully to decrease ischemia-reperfusion injury in several organ systems. We evaluated the efficacy of poly (ADP-ribose) polymerase inhibitors on biochemical changes in testicular ischemia-reperfusion injury. MATERIALS AND METHODS: Adult male Wistar rats were divided into 9 groups of 6 each. One group served to determine baseline values of biochemical parameters, 1 that underwent sham operation served as a control, 1 underwent 2 hours of testicular torsion and 4 hours of detorsion, 2 received pretreatment with vehicle (saline or dimethyl sulfoxide) before detorsion and 4 received pretreatment with the poly (ADP-ribose) polymerase inhibitor nicotinamide, 3-aminobenzamide, 1,5-dihydroxyisoquinoline or 4-amino-1,8-naphthalimide before detorsion. Lipid peroxidation products, nitric oxide content and myeloperoxidase activity, an indicator of neutrophil accumulation, were assessed in testicular and renal tissues. RESULTS: Testicular torsion-detorsion caused a significant increase in lipid peroxidation products, nitric oxide content and myeloperoxidase activity in ipsilateral testes (p <0.01) but not in the contralateral testes or kidneys. Animals treated with poly (ADP-ribose) polymerase inhibitors had a significant decrease in these biochemical parameters compared with vehicle treated animals (p <0.01). CONCLUSIONS: These data emphasize that poly (ADP-ribose) polymerase may have a role in testicular damage caused by ischemia-reperfusion and the inhibition of poly (ADP-ribose) polymerase may be a novel approach to therapy for ischemia-reperfusion injury of the testis.

The protective role of caffeic acid phenethyl ester (CAPE) on testicular tissue after testicular torsion and detorsion.

Testicular artery occlusion causes an enhanced formation of reactive oxygen species, which contributes to the pathophysiology of tissue damage. Here, we have investigated the effects of caffeic acid phenethyl ester (CAPE), a new antioxidant and antiinflammatory agent, in rats subjected to testicular torsion/detorsion (T/D). Thirty-five male rats were divided into four groups: sham operation group ( n=8), torsion group ( n=9), T/D+saline group ( n=9) and T/D+CAPE group ( n=9). Rats, except the sham operation group, were subjected to left unilateral torsion (720 degrees rotation in the clockwise direction) without including the epididymis. After torsion (2 h) and detorsion (4 h) periods, rats were sacrificed and bilateral orchidectomy was performed. Testis tissues were washed with cold saline solution, cut into small pieces with scissors, placed into glass bottles and homogenised in four volumes of ice-cold Tris-HCl buffer. Clear supernatant fluid was used for biochemical analyses. Treating rats with CAPE (applied at 10 micro mol/kg, 30 min prior to T/D) attenuated the testicular injury, as well as the increase in the tissue levels of myeloperoxidase and thiobarbituric acid-reactant substances (TBARS) caused by T/D in the testis. Testis tissues showed a significant increase in glutathione peroxidase (GSH-Px) activity compared to the torsion group when CAPE was applied. Taken together, our results clearly demonstrate that CAPE treatment exerts a protective effect on testicular T/D, and part of this effect may be due to inhibiting the neutrophil-mediated cellular injury.

Serum creatine kinase enzyme levels in the early diagnosis of spermatic cord torsion.

Delay in the diagnosis of spermatic cord torsion (SCT) is still a significant cause of testicular loss in children. The aim of this experimental study was to assess the diagnostic value of serum creatine kinase (CK) in the early period following SCT. Forty male rats were assigned randomly into five similar groups: group A, control; group B, sham, right testis exposed, manipulated, and blood sampling at 6th h; group C, right SCT, blood sampling at 2nd h; group D, right SCT, blood sampling at 4th h; and group E, right SCT, blood sampling at 6th h. Ck and its isoenzymes were measured in the sera of all animals. All testes were removed and examined histopathologically. Significant increases in serum CK levels compared to control and sham groups were observed at 4 and 6 h following SCT. The major increase in CK was observed in the CK-MM isoenzyme fraction. Histologic pictures showed varying degrees of edema, vascular congestion, and hemorrhage in the testicular tissue, but no necrosis in any of the study groups. These results showed that serum CK levels in rats in the early period following SCT increase significantly before necrosis of testicular tissue. This may be of value as a diagnostic test, to corroborate findings from clinical studies.

Alteraçöes enzimáticas na torçäo do cordäo espermático: estudo experimental no çäo / Enzymatic changes in the spermatic cord torsion: experimental study in dogs

Seis cäes foram submetidos a torçäo do cordäo espermático e determinou-se a dosagem em diferentes intervalos das enzimas creatinina fosfoquinase e desidrogenase lática. Um aumento significante da creatinina fosfoquinase foi observado em 67% dos animais. Estudos clínicos deveräo ser efetuados para avaliar a possibilidade da aplicaçäo do método

[Experimental torsion of the spermatic cord. Histological and histochemical studies (author's transl)]. / Experimentelle Hodentorsion. Histologische und enzymhistochemische Untersuchungen.

In 40 male adult rats on unilateral torsion of the spermatic cord was initiated by operation. In the first group (4 x 5 animals) the testes were removed after 3, 6, 12, 24 hours. The treatment of the animals in the second group (4 x 5 animals) was untwisting of the torsion after 3, 6 12, 24 hours. Three months later both testes were removed. Histological and enzymehistochemical results are as follows: 1. The extend of acute damage to the affected testis depends on the length the torsion lasted. 2. The delay of time of untwisting the testis probably does not influence the degree of later regeneration. 3. Even short duration of torsion of the affected testis results in damage to the contralateral testies.