Testicular Ischemia-reperfusion Injury Research Articles

Quercetin has a protective role on histopathological findings on testicular ischaemia-reperfusion injury in rats

We investigated the effects of quercetin on pathological findings on testicular ischaemia-reperfusion (I/R) injury in rats. Twenty-four male Wistar albino rats were randomly assigned into four groups: Group 1, control (n = 5); Group 2, sham (n = 4); Group 3, I/R (n = 8); and Group 4, I/R + quercetin (n = 7). Bilateral testicular artery and vein were occluded for 1 h, followed by reperfusion in I/R and I/R + quercetin animals. Quercetin (20 mg kg(-1) per day) was administrated once daily by gavage to Group 1 and Group 4, respectively, after reperfusion. At the end of the study, bilateral orchiectomies were performed for histopathologic examination. The tissue damage was evaluated with light microscopy. Normal inter-stitium and seminiferous tubules were observed in control group. In the sham group, rats were seen minimal oedema around the seminiferous tubules and congested vascular structures. In Group 3, oedema, vascular congestion and haemorrhage between seminiferous tubules were observed. In Group 4, histopathologic features were markedly less than Group 3 (P = 0.03). Our study demonstrated that quercetin seems to have a protective effect on testis histopathology in rats with testicular I/R.

Protective Effect of Ischemic Postconditioning Against Testicular Ischemia-Reperfusion Injury in Rabbits

PurposeTo investigate the protective effect of ischemic postconditioning (IP) against testicular ischemia/reperfusion (I/R) injury of different degrees in rabbits.Material & MethodsA total of 42 rabbits were randomly divided into seven groups: control group, different degree I/R group (R1, R2, R3),and different degree IP group (P1, P2, P3), six in each group. nder ultrasound monitoring, made of different degree of testicular ischemia model. Group R1 and group P1 showed homogeneous echo and flow signals slightly decreased, group R2 and P2 showed heterogeneous echo and flow obviously decreased, group R3 and P3 showed radial or fragmental low echo areas and flow disappeared. Before the long time reperfusion, group I/R had direct perfusion, group IP was subjected to perfusion for 30 s and then ischemia for 30 s, repeated three times. After 3 days of breeding, malonaldehyde (MDA) and superoxide dismutase (SOD) levels, and histology were measured.ResultsThe levels of MDA and SOD had significant differences between group R1 and P1 (p < 0.05), and between group R2 and P2 (p < 0.05), but there were no significant differences between group R3 and P3 (p > 0.05). Johnsen's score and apoptosis index had significant differences in group R1 and P1 (p < 0.05), but there were no significant differences in group R2 and P2 (p > 0.05), and in group R3 and P3 (p > 0.05).ConclusionIP can attenuate testicular damage induced by I/R, but it is restricted by the degree of ischemia. Scrotal ultrasound can be used to predict the effect of IP against testicular IR injury. PurposeTo investigate the protective effect of ischemic postconditioning (IP) against testicular ischemia/reperfusion (I/R) injury of different degrees in rabbits. To investigate the protective effect of ischemic postconditioning (IP) against testicular ischemia/reperfusion (I/R) injury of different degrees in rabbits. Material & MethodsA total of 42 rabbits were randomly divided into seven groups: control group, different degree I/R group (R1, R2, R3),and different degree IP group (P1, P2, P3), six in each group. nder ultrasound monitoring, made of different degree of testicular ischemia model. Group R1 and group P1 showed homogeneous echo and flow signals slightly decreased, group R2 and P2 showed heterogeneous echo and flow obviously decreased, group R3 and P3 showed radial or fragmental low echo areas and flow disappeared. Before the long time reperfusion, group I/R had direct perfusion, group IP was subjected to perfusion for 30 s and then ischemia for 30 s, repeated three times. After 3 days of breeding, malonaldehyde (MDA) and superoxide dismutase (SOD) levels, and histology were measured. A total of 42 rabbits were randomly divided into seven groups: control group, different degree I/R group (R1, R2, R3),and different degree IP group (P1, P2, P3), six in each group. nder ultrasound monitoring, made of different degree of testicular ischemia model. Group R1 and group P1 showed homogeneous echo and flow signals slightly decreased, group R2 and P2 showed heterogeneous echo and flow obviously decreased, group R3 and P3 showed radial or fragmental low echo areas and flow disappeared. Before the long time reperfusion, group I/R had direct perfusion, group IP was subjected to perfusion for 30 s and then ischemia for 30 s, repeated three times. After 3 days of breeding, malonaldehyde (MDA) and superoxide dismutase (SOD) levels, and histology were measured. ResultsThe levels of MDA and SOD had significant differences between group R1 and P1 (p < 0.05), and between group R2 and P2 (p < 0.05), but there were no significant differences between group R3 and P3 (p > 0.05). Johnsen's score and apoptosis index had significant differences in group R1 and P1 (p < 0.05), but there were no significant differences in group R2 and P2 (p > 0.05), and in group R3 and P3 (p > 0.05). The levels of MDA and SOD had significant differences between group R1 and P1 (p < 0.05), and between group R2 and P2 (p < 0.05), but there were no significant differences between group R3 and P3 (p > 0.05). Johnsen's score and apoptosis index had significant differences in group R1 and P1 (p < 0.05), but there were no significant differences in group R2 and P2 (p > 0.05), and in group R3 and P3 (p > 0.05). ConclusionIP can attenuate testicular damage induced by I/R, but it is restricted by the degree of ischemia. Scrotal ultrasound can be used to predict the effect of IP against testicular IR injury. IP can attenuate testicular damage induced by I/R, but it is restricted by the degree of ischemia. Scrotal ultrasound can be used to predict the effect of IP against testicular IR injury.

Protective effect of rutin on testicular ischemia-reperfusion injury

Purpose The pathophysiology of testicular torsion-detorsion is an ischemia-reperfusion injury caused by overgeneration of reactive oxygen species (ROS). This study aimed to investigate the effect of rutin, a well-known antioxidant, on testicular ischemia-reperfusion injury. Methods Sixty male Sprague-Dawley rats were randomly divided into 3 groups, each containing 20 rats. Rats in the control group underwent a sham operation of the left testis. In the torsion-detorsion group, the left testis was rotated 720° for 2 hours. Rats in the treatment group received the same surgical procedure as the torsion-detorsion group, but rutin was administered intravenously at the time of detorsion. Bilateral orchiectomy was performed on half of the rats in each experimental group at 4 hours after detorsion for measurement of malondialdehyde, an indicator of intratesticular ROS content, and for evaluation of superoxide dismutase and catalase, which are endogenous antioxidant enzymes. Orchiectomy was performed on the remaining rats at 3 months after detorsion for analysis of testicular spermatogenesis. Results Unilateral testicular torsion-detorsion caused a significant increase in malondialdehyde level and caused significant decreases in superoxide dismutase, catalase activities, and spermatogenesis in ipsilateral testes. The rats treated with rutin had a significant decrease in malondialdehyde level and had significant increases in superoxide dismutase, catalase activities, and spermatogenesis in ipsilateral testes, compared with torsion-detorsion group. Conclusions Rutin protects testes from ischemia-reperfusion injury. The protective effect of rutin may be caused by scavenging ROS by increasing superoxide dismutase and catalase activities.

Does propofol prevent testicular ischemia–reperfusion injury due to torsion in the long term?

Our aim was to investigate the long-term preventive effect of propofol on testicular ischemia-reperfusion injury in a rat model. Twenty-four adult male Sprague-Dawley rats were randomly divided into four groups (n = 6 for each group), control, sham-operated, torsion/detorsion (T/D) and T/D + propofol. Testicular ischemia was achieved by twisting the left testis 720° clockwise for 2 h. Half an hour before detorsion, 50 mg/kg propofol was given intraperitoneally to the T/D + propofol group. Ipsilateral orchiectomies to determine mean testicular weights and histopathological examination according to Johnsen's mean testicular biopsy score criteria were performed 30 days after surgical procedure in all groups. Mean testicular weights were 1.57 ± 0.12 g in group I, 1.59 ± 0.36 g in group II, 0.84 ± 0.20 g in group III and 0.87 ± 0.29 g in group IV. Mean testicular weights decreased significantly in the T/D groups, but no improvement in testicular weight was observed with propofol administration (p 0.9372). Similarly, the Johnsen's mean testicular biopsy scores of the T/D groups were lower than those of the control and sham-operated groups, but no positive effect was determined with the administration of propofol in the T/D groups (p 0.1797). Our results showed that there is no apparent long-term therapeutic potential attendant on using propofol in the treatment of testicular ischemia-reperfusion injury caused by testis torsion.

The effects of short-interval postconditioning in preventing testicular ischemia-reperfusion injury in rats

AimTesticular torsion can lead to testicular damage. During reperfusion, tissue damage is more severe. The aim of this study was to investigate the protective effect of short-interval postconditioning and determine the optimal time of reperfusion for postconditioning. Materials and MethodsThirty-five adult male rats were divided into 5 subgroups: Sh (sham operated), TD (torsion + detorsion), PC5 (torsion + postconditioning 5 seconds), PC10 (torsion + postconditioning-10 seconds), PC20 (torsion + postconditioning 20 seconds). Torsion was created by rotating the left testis counterclockwise 1080° and the testis fixed to the scrotum with 3 sutures. Torsion was maintained for 4 hours. The testicular artery was visualized, and before detorsion of the testis, an atraumatic vessel clamp was applied to prevent reperfusion in all study groups. Then, detorsion of the testis was performed. In the TD group, the clamp was released just after detorsion; in the PC5 group, the clamp was released for 5 seconds and closed for 10 seconds (10 times); in the PC10 group, the clamp was released for 10 seconds and closed for 10 seconds (10 times); and in the PC20 group, the clamp was released for 20 seconds and closed for 10 seconds (10 times). Then, all testes were reperfused for a 1-hour period in all study groups. After this period, the rats were sacrificed, and the left testes were removed and evaluated histopathologically and biochemically. The Mann-Whitney U test was used for statistical analyses. ResultsTissue malondialdehyde levels were 79.3 ± 10.6, 231.7 ± 102.3, 71.3 ± 12.6, 73.8 ± 13.7, and 124.3 ± 48.0 nmol/g tissue in the Sh, TD, PC5, PC10, and PC20 groups, respectively. Tissue malondialdehyde levels were significantly lower in the PC5 and PC10 groups (P < .05) compared to the other groups. However, mean histopathologic grade was lower in all postconditioning groups compared to the control group, but the difference was significant only in the PC5 group (P < .05). ConclusionWe conclude that short-interval postconditioning can reduce reperfusion injury in ischemic tissue and the optimal mode of short-interval postconditioning is 5 seconds × 10 times. This technique seems easily applicable, and a similar technique may be used during testicular surgery.

Protective effect of sivelestat, a neutrophil elastase inhibitor, on ipsilateral and contralateral testes after unilateral testicular ischaemia–reperfusion injury in rats

To investigate the effect of a neutrophil elastase inhibitor, sivelestat sodium hydrate, on testicular ischaemia-reperfusion (IR)-injury. Eight-week-old male Sprague-Dawley rats were divided into four groups: sham-operated control rats; IR rats (group IR); and IR rats that received intra-abdominal administration of 15 mg/kg or 60 mg/kg sivelestat (group IR15 and group IR60, respectively). Right testicular vessels were clamped for 90 min in groups IR, IR15 and IR60. Sivelestat had been administered 45 min after the induction of the ischaemia in groups IR15 and IR60. In subpopulations of IR, IR15 and IR60 rats, reperfusion was performed after ischaemia for 2 h (groups IR-A, IR15-A and IR60-A, respectively) or 48 h (groups IR-B, IR15-B and IR60-B, respectively). At the end of the reperfusion period, blood samples were aspirated from both spermatic veins of each rat and testosterone was evaluated. Then both testes from all rats were collected and tissue levels of malondialdehyde (MDA), myeloperoxidase (MPO), and heat-shock protein-70(HSP-70) were evaluated. Testicular tissue samples were also processed for histological evaluation and TUNEL staining. MDA, MPO and HSP-70 levels in the ischemic testis were significantly higher in the IR group compared with the control group. MDA and HSP-70 in the contralateral testis were significantly higher in the IR group compared with the control group. Bilateral testosterone levels were lower in all rat groups in comparison with the control group. Bilateral testicular samples in group IR showed extensive histopathologic degenerative alterations and increased percentage of apoptotic cells. Sivelestat treatment lowered the MDA concentration and the percentage of apoptotic cells bilaterally and ameliorated the testicular histological pattern bilaterally. Unilateral testicular ischaemia causes significant contralateral testicular damage. Sivelestat may be a novel adjunct tool for reducing oxidative stress and partially preventing bilateral testicular damage.

Effects of 3-aminobenzamide on unilateral testicular ischemia–reperfusion injury: What is the role of PARP inhibition?

The therapeutic effects of poly(adenosine diphosphate-ribose) polymerase inhibition by 3-aminobenzamide (3-AB) were investigated in testicular ischemia-reperfusion (I/R) injury, using sperm analysis and histopathological and biochemical examinations, including superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) activities and reduced glutathione (GSH) levels. Male rats were divided into 3 groups: sham (n = 12), I/R (n = 12), and I/R with 3-AB (I/R-3-AB) (n = 12). The left testicular artery was occluded for 1 h, followed by 24 h (for biochemical and histopathological examinations) and 30 days (for sperm analysis) of reperfusion. 3-AB treatment intraperitoneally 10 min prior to and 1 h after reperfusion increased the I/R-induced decrease in sperm motility in both testes and reduced the increased abnormal sperm rates in the ipsilateral testis. However, 3-AB treatment failed to prevent the I/R-induced decrease in sperm concentration in both testes. SOD and CAT activities did not change in any group. GSH-Px activity and GSH levels were increased by I/R. 3-AB treatment reversed the I/R-induced increase in GSH-Px activity, similar to the level in sham rats, but did not alter GSH levels. 3-AB treatment significantly increased the I/R-induced decrease in histopathologic score. In conclusion, 3-AB treatment has potential biochemical and histopathological benefits beyond improving sperm quality and may have the potential to decrease damage from testicular torsion.

Protective Effect of Thymoquinone in Experimental Testicular Torsion

Objectives: To investigate the protective role of thymoquinone (TQ) on unilateral testicular ischemia-reperfusion (I/R) injury in mice. Materials and Methods: Experiments were performed on male C57BL/6 mice (8 weeks old, 20–25 g). The animals were divided into 3 groups including 6 mice in each group: control (sham), torsion/detorsion (TD) and TD+TQ. Mice, except the sham-operated group, were subjected to left unilateral torsion (720° rotation in the clockwise direction). The experiments were finished after sham operation time for controls, 120 min torsion and 240 min detorsion for the other groups. In the TD+TQ group 10 mg TQ was injected intraperitoneally 30 min before detorsion. Results: In the TD group total oxidative stress (TOS), oxidative stress index (OSI) and malondialdehyde (MDA) levels were higher than in the controls. TQ treatment decreased MDA, TOS and OSI values, but did not affect the total antioxidant capacity and myeloperoxidase activity in the TD+TQ group. Upon histological examination, mice in the TD group displayed moderate-to-severe disruption of the seminiferous epithelium. Treatment with TQ resulted in significantly reduced histological damage associated with I/R injury. Conclusion: Our results suggested that TQ treatment may have a protective effect on testicular I/R injury.

Cold Ischemia Time Influences Spermatogenesis in a Testicular Ischemia/Reperfusion Injury Model

Objective To evaluate the influence of cold ischemia time on spermatogenesis in a rabbit model of testicular ischemia-reperfusion (I/R) injury. Material and Methods The testicular I/R model was established in 24 male white rabbits. The left testes were preserved using HC-A solution at 0°C to 4°C. Cold ischemia time was 1, 2, 4, and 6 hours. The right testes without vascular occlusion were used as autologous controls. Twenty-four hours after reperfusion, the animals were sacrificed, and samples were obtained at bilateral orchiectomy. Another 8 normal testes were used as normal controls. Testicular tissue Johnsen score, malondialdehyde concentration, and apoptosis index were used to evaluate spermatogenesis. Results The Johnsen score decreased and the apoptosis index increased with the duration of cold ischemia time in the I/R groups. The malondialdehyde concentration in the I/R groups was significantly higher than the sham and normal groups, and was highest at 4 hours of cold ischemia time. Conclusion Testicular I/R injury is highly related to cold ischemia time. In rabbit models, testis transplantation is best performed within 4 hours of cold ischemia with traditional hypothermic protection.

Protective effect of ischaemic post-conditioning on ipsilateral and contralateral testes after unilateral testicular ischaemia-reperfusion injury

The aim of this study was to investigate the effect of ischaemic post-conditioning (IPostC) against ischaemia-reperfusion (IR) injury on bilateral testes after unilateral testicular ischaemia in the rat. Eight-week-old male Sprague-Dawley rats were divided into control group; IR group (60 min ischaemia-24 h reperfusion); IPostC1 × 10 group (60 min ischaemia followed by one cycle of 10 sec reperfusion-10 sec ischaemia; then 24 h reperfusion); IPostC3 × 10 group (three cycles of 10 sec reperfusion-10 sec ischaemia; then 24 h reperfusion); IPostC5 × 10 group (five cycles of 10 sec reperfusion-10 sec ischaemia; then 24 h reperfusion) and IPostC3 × 30 group (three cycles of 30 sec reperfusion-30 sec ischaemia; then 24 h reperfusion). In the IR and IPostC groups, the right testicular vessels were clamped using a special vascular clip. Malondialdehyde (MDA) and myeloperoxidase (MPO) levels were measured in testicular tissue samples bilaterally. Additionally, bilateral testicular tissue samples were processed for histological evaluation including haematoxylin-eosin, 4-hydroxy-2-nonenal (4-HNE) and TdT-mediated dUTP Nick End Labelling (TUNEL) staining. The levels of MDA and MPO as well as the positive cells per seminiferous tubule in TUNEL and 4-HNE stain in bilateral testes from the IR group were significantly higher compared with the control group. IPostC3 × 30 protocol significantly ameliorated the aforesaid parameters in both testes compared with the IR group. For the first time, we have demonstrated that IPostC protects both testes after unilateral testicular ischaemia-reperfusion. IPostC3 × 30 protocol offered the most effective protection.

The effects of N-acetylcysteine on testicular damage in experimental testicular ischemia/reperfusion injury

The aim of this study to evaluate the effects of N-acetylcysteine (NAC) on testicular damage in a rat testicular ischemia-reperfusion (I/R) injury model. Thirty male Wistar albino rats were divided into five groups. Group 1: sham control, Group 2: torsion (T), Group 3: torsion/detorsion (T/D), Group 4: the early NAC treatment plus T/D, 20 mg/kg of NAC was given intravenously 60 min before detorsion; Group 5: the late NAC treatment plus T/D, 20 mg/kg of NAC was given intravenously 5 min before detorsion. After torsion (2 h) and detorsion (2 h), bilateral orchiectomies were performed to determine the tissue levels of malondialdehyde (MDA) or more exactly thiobarbituric acid reactive substance (TBARS), myeloperoxidase activity and histopathological changes. The most significant increase in the mean TBARS level and decrease in the mean seminiferous tubular diameter, germinal epithelial cell thickness values in bilateral testes were observed in T/D group rather than other groups. TBARS levels of early NAC treatment group were significantly lowered and histological parameters of spermatogenesis were significantly improved in bilateral testes when compared with T and T/D groups. Our results suggest that the early administration of NAC may have a protective effect in the rat experimental testicular T/D models.

The role of vascular endothelial growth factor in ischemia-reperfusion injury of testis

OBJECTIVE: Testicular torsion causes ischemia-reperfusion injury of testis and leads to male factor infertility. Its injury initiates a pathophysiologic cascade. In recent studies, protective effect of VEGF has been shown in various forms of ischemia- reperfusion injury. However, no study has investigated the role of VEGF in testicular ischemia-reperfusion injury. The aim of our study was to investigate the time course expression of VEGF, VEGF-receptor (R)1, VEGF-R2, inducible and endothelial nitric oxide synthase (iNOS and eNOS) and to evaluate the effect of VEGF in ischemia-reperfusion injury of testis. DESIGN: Animal study. MATERIALS AND METHODS: Using 8-week-old SD rat, surgery for making unilateral experimental torsion was performed. After 1 hour of torsion, the testis was counter-rotated back to the natural position and reinserted into the scrotum. Bilateral castrations were performed in various time courses after reperfusion. Western bolt analysis and immunohistochemical evaluation were used to determine the activation of VEGF, iNOS, and eNOS. VEGF, VEGF-R1 and -R2 mRNAs were measured by reverse transcription PCR (RT-PCR) and quantitative real-time PCR (QRT-PCR) analysis. RESULTS: In torsion side testis, the expression of VEGF protein and mRNA significantly increased in a time dependent manner since 3 to 120 hour after reperfusion. Although the expression of VEGF-R1 mRNA was increased in a similar way, which of VEGF-R2 mRNA could not be detected. The increase of VEGF protein expression was observed in testicular vascular endothelial cells and seminiferous tubular endothelial cells. Significant activation of iNOS and eNOS were investigated at 12 and 24 hours after reperfusion. CONCLUSIONS: Our study is the first report in the literature to show the time course expression of VEGF in torsion caused ischemia-reperfusion injury testis. Further investigations, such as VEGF infusion study, may lead to a novel approach for the therapy of ischemia-reperfusion injury of the testis.

Protective effects of L-carnitine upon testicular ischemia-reperfusion damage in rats

To investigate the protective effects of L-carnitine upon testicular ischemia-reperfusion injury in rats. Sprague-Dawley rats were divided into 3 groups (n = 10). In those animals undergoing unilateral testicular torsion, right testes were rotated 720 degrees for 2 h. Sham operated group served as a control group. Torsion group underwent 2 h torsion and saline was injected intraperitoneally at 30 min pre-detorsion. Treatment group underwent similar torsion but L-carnitine (500 mg/kg) was infused intraoperatively. The right testes of 5 animals in each group were excised after 4 h reperfusion for measuring the levels of malondialdehyde (MDA) and heat shock protein 70 (HSP70), evaluation of activities of antioxidant enzyme including superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx). Histopathological changes and germ cell apoptosis indices (AI) were determined at 24 h post-detorsion in right testes of the remaining 5 animals in each group. The mean number of apoptotic nuclei per tubule cross section and the malondialdehyde level were significantly lower in treatment group as compared with torsion group [AI( 6.87 +/- 2.47) vs (17.13 +/- 3.56), MDA (160 +/- 15) vs (199 +/- 15) nmol/g]. Activities of antioxidant enzyme and the level of HSP70 were significantly higher in treatment group than those in torsion group [SOD (1638 +/- 153) vs (1078 158) U/g, CAT (317 +/- 28) vs (188 +/- 33) U/g, GPx (667 +/- 94) vs (311 +/- 65) U/g, HSP70 (0.87 +/- 0.13) vs (0.25 +/- 0.04)]. The pathological damage of testes in the treatment group was lighter than that in the torsion group (all P < 0.05). The administration of L-carnitine exerts a beneficial effect upon testicular ischemia-reperfusion injury. This effect may be achieved through an induced expression of HSP70.

Involvement of the bax and bcl-2 system in the induction of germ cell apoptosis is correlated with the time of reperfusion after testicular ischemia in a rat model

The objective of this study was to examine the relationship between time of reperfusion and bax/bcl-2-dependent germ cell apoptosis after testicular ischemia-reperfusion injury in the rat. In ischemic testis, bax/bcl-2 ratio did not change significantly, and the elevation of germ cell apoptosis was not marked; in the contralateral testis, germ cell apoptosis increased after 6 hours of reperfusion, achieved statistical significance after 24 hours, and decreased after 72 hours of reperfusion and was initiated by decreased bcl-2 messenger RNA levels and elevated bax/bcl-2 ratio within the first 6 hours of reperfusion.

Effect of melatonin on epididymal sperm quality after testicular ischemia/reperfusion in rats

To determine the effect of melatonin, a pineal secretory product that prevents testicular ischemia/reperfusion (IR) injury through its antioxidative properties, on epididymal sperm quality in a rat testicular IR injury model. Experimental study. University pharmacology laboratory. Fifty-six 8-week-old male Wistar albino rats. Left testicular artery and vein occluded for 1 hour; before the bilateral orchiectomy, the organ was allowed to reperfuse 30 days. Melatonin (10 mg/kg IP) or vehicle (1% ethanol in saline) was administrated for 10 minutes before reperfusion and for 1 hour after reperfusion. After 24 hours of reperfusion, the rats were decapitated, and the testicular tissue samples were obtained for histologic examination. In addition, after 30 days of reperfusion, the epididymal sperm concentration, motility, and abnormal sperm rates were determined in the sperm collected from the epididymis. A statistically significant decrease in sperm concentration resulted from IR as well as an increase in sperm abnormalities, but the sperm motility did not change. Melatonin treatment did not prevent the IR-induced reduction in sperm concentration. However, melatonin treatment statistically significantly decreased the sperm abnormalities when compared with the IR injured samples. Melatonin may improve sperm morphology for a protective effect in IR-induced testicular injury.

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

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. 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. 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. Ginkgo biloba, as a free radical scavenger, seems to have a protective role against apoptosis in testicular ischemia reperfusion injury.

Mitogen-activated protein kinase 3/mitogen-activated protein kinase 1 activates apoptosis during testicular ischemia–reperfusion injury in a nuclear factor-κB-independent manner

Nuclear factor kappa-B (NF-κB), mitogen-activated protein kinase3/MAPK1 and MAPK8 are involved in testicular ischemia reperfusion injury (testicular-I/R). NF-κB knock-out mice (KO) subjected to testicular-I/R have a reduced testicular damage, blunted MAPK8 activation and enhanced MAPK3/MAPK1 activity. To better understand the role of MAPK3/MAPK1 up-regulation during testicular-I/R, we investigated the effects of PD98059, an inhibitor of MAPK3/MAPK1, in KO mice during testicular-I/R. KO and wild-type (WT) animals underwent 1 h testicular ischemia followed by 24 h reperfusion or a sham testicular-I/R. Animals received either PD98059 (5 mg/kg/ip) or its vehicle. MAPK3/MAPK1, BAX, caspase-3 and -9 and TNF-α expression were assessed along with histological examination and an immunostaining for protein of apoptosis. Testicular-I/R caused a greater increase in MAPK3/MAPK1 in KO than in WT animals in both testes. KO mice had a lower expression of the apoptotic proteins and TNF-α as well as reduced histological damage compared to WT. Immunostaining confirmed the lower expression of BAX in the Leydig cells of KO mice. Administration of PD98059, abrogated MAPK3/MAPK1 expression and slightly reduced TNF-α but did not improve or reverse the histological damage in KO. PD98059 significantly reduced the histological damage in WT mice and markedly reduced the apoptotic proteins in KO and WT mice. These results suggest that testicular-I/R triggers also a pathway of organ damage involving MAPK3/MAPK1, TNF-α, BAX, caspase-3 and -9 that activates an apoptotic machinery in an NF-κB independent manner. These findings should contribute to better understand testicular torsion-induced damage.

Gradual detorsion of torsioned rat testis attenuates ischemia reperfusion injury

Aim This study was designed to investigate effect of gradual detorsion on testicular ischemia reperfusion injury. Materials and Methods A total of 21 male rats were divided into 3 groups, each containing 7 rats. Torsion was created by rotating the left testis 720° in a clockwise direction. Group 1 underwent sham operation. Group 2 (sudden detorsion) served as a torsion/detorsion group, receiving 2 hours torsion and 2 hours detorsion. In group 3, 360° detorsion was done for 20 minutes after 720° torsion for 2 hours. Then, testis was done full detorsion for 100 minutes. At the end of the experiments (fourth hour), left orchiectomy was performed to measure the tissue levels of malondialdehyde (MDA), superoxide dismutase, and glutathione peroxidase and to perform histologic examination in testes. Results The MDA levels of testis tissues were significantly increased in the sudden detorsion group as compared with the sham group. We found decrease of the MDA level in gradual detorsion group, but it was not a statistically significant amount. Significant decrease was found in the superoxide dismutase and glutathione peroxidase activities in the sudden detorsion group as compared with the sham and gradual detorsion groups. Histologic examinations were in accordance with the testicular tissue MDA levels. Conclusion In the light of our biochemical and histopathologic findings, we can say that gradual detorsion has a trend to decrease the degree of testicular reperfusion injury in the rat torsion/detorsion model.

Effects of melatonin on spermatogenesis and testicular ischemia-reperfusion injury after unilateral testicular torsion-detorsion

Purpose This study aimed to determine the effect of melatonin in preventing ischemia-reperfusion (I-R) injury-induced tissue damage and on spermatogenesis after experimental testicular torsion (TT). Methods Forty peripubertal rats were divided into 4 groups each containing 10 rats: control (C), sham (S), torsion plus detorsion (TD), and torsion plus melatonin (M). The left testes were rotated 720° for 6 hours and detorsed for 6 hours thereafter. Serum inhibin B (IB) levels were measured in blood samples taken from all groups. Left orchiectomies were performed to determine the tissue levels of Johnsen's scores (JS) and malondialdehyde (MDA). Results Serum IB levels in the S and TD groups were significantly lower compared with that in the C group, whereas they were higher in the M group compared with the TD group. The MDA levels were significantly lower in the C, S, and, M groups compared with the TD group. Johnsen's scores were significantly higher in the C, S, and M groups compared with the TD group. Conclusions Our results suggested that melatonin is a potent antioxidant agent in preventing testicular I-R injury, as shown by increased IB levels and JS.

Protective effect of trapidil on long-term histologic damage in a rat model of testicular ischemia-reperfusion injury

Trapidil is an antianginal compound with a broad spectrum of pharmacological activities. In recent years, it has been used successfully to decrease ischemia-reperfusion injury in several organ systems. We evaluated the effect of trapidil on the long-term histologic damage in testicular ischemia-reperfusion injury. Adult male Wistar rats were divided into three groups of six rats each. One group underwent 2 h of testicular torsion; one received pretreatment with trapidil before detorsion; and one group underwent sham operation. All rats underwent bilateral orchiectomy 60 days after the experiment. The mean seminiferous tubular diameter, germinal epithelial cell thickness, and mean testicular biopsy score were determined by histological examination of each testis. Testicular torsion-detorsion caused a significant decrease in the mean seminiferous tubular diameter, germinal epithelial cell thickness, and mean testicular biopsy score in the ipsilateral testes, but not in the contralateral testes. The animals treated with trapidil had a significant increase in these histological parameters as compared to the torsion-detorsion group. Trapidil administration before reperfusion may have the potential to decrease the long-term histologic damage that occurs after experimental testicular torsion. Trapidil is used as an antianginal drug and additional clinical studies are required to elucidate the protective role of trapidil in patients with testicular torsion.