Effects of co-administration of dopamine and vitamin C on ischaemia-reperfusion injury after experimental testicular torsion-detorsion in rats

The objective of this study was to investigate the effects of dopamine as vasodilator, vitamin C as an antioxidant and combined administration of them on ischaemia-reperfusion (I-R) injury following testicular torsion (TT). Thirty adult male rats were divided into six groups each containing five rats. Testicular ischaemia was achieved by twisting the left testis for 4 h. Group 1 was for determination of the basal values. Group 2 had 4 h TT. Group 3 had 4 h TT and was then treated with dopamine. Group 4 had 4 h TT and was then treated with vitamin C. Group 5 had 4 h TT and was then treated with dopamine and vitamin C. Group 6 was designed as a sham operated group. Testicular torsion caused a significant decrease in the percentage of spermatogenesis and seminiferous tubules diameters compared with the control and sham groups. Administration of dopamine, vitamin C and their combination increased above mentioned parameters and decreased serum malondialehyde levels significantly. However, vitamin C had better results than the other treatments (P < 0.05). In conclusion, a potent antioxidant like vitamin C was found to be more effective than increasing blood flow by a vasodilator like dopamine on improving I-R injury following TT.     

Metformin decreases testicular damages following ischaemia/reperfusion injury in rats

The effects of metformin on a testicular torsion injury in adolescent rat testis after I/R were evaluated in the present study. Forty adolescent rats were divided into five groups with eight rats per group: a control group; a sham-operated group; an ischaemia group, where torsion was applied for 4 hr and testis was examined immediately after detorsion; an I/R group, where torsion was applied for 4 hr and the testis was examined 4 hr after detorsion; and an I/R + M group, where the metformin (300 mg/kg) administration was added to the identical procedures used for the I/R group. Spermatogenesis, basal membrane integrity and cleaved caspase-3 expression were assessed. The I/R + M group had a significantly higher Johnsen score than the I/R group (7.9 ± 0.1 vs. 7.5 ± 0.2; p < .001; F-value = 14.2). Failure of basal membrane integrity was highest in the ischaemia group (45 ± 5) compared to the other groups (control group, 20 ± 5; sham-operated group, 16.6 ± 2.8), but not different between the I/R + M (31.6 ± 12.5) and the I/R groups (25 ± 3.5). Cleaved caspase-3 expression was highest in the ischaemia group (73.5 ± 0.7), and significantly lower in the I/R + M group (33.4 ± 0.9) than the I/R group (58.5 ± 0.2; p < .05; F-value = 7.6). Metformin decreases testicular damage by exerting protection against the harmful effects of I/R on spermatogenesis and alleviating apoptosis in adolescent rat testis.     

Protective effect of liraglutide on experimental testicular ischaemia reperfusion in rats

This study was performed to evaluate the effect of liraglutide on experimental testicular ischaemia reperfusion in rats in terms of biochemistry, histopathology and immunohistochemistry. A total of 28 male Wistar-Albino rats were divided randomly into 4 groups: control (7), sham (7), ischaemia-reperfusion (7) and ischaemia-reperfusion + liraglutide (7). Biochemically, Nitric Oxide, Malondialdehyde, Superoxide dismutase, Glutathione peroxidase and Catalase levels were measured in the testis. Apoptosis protease activating factor-1 and inducible nitric oxide synthase activity were evaluated immunohistochemically as well. Statistical analyses were made via the Kruskal–Wallis and Mann–Whitney U tests. In the reperfusion group, CAT and SOD values were increased (p > .05), NO and MDA values were decreased (p < .05) after administration of liraglutide. In addition, GPx values were significantly increased in ischaemia reperfusion + liraglutide administered group compared to reperfusion group (p < .05). Apaf-1 and iNOS activity were significantly decreased with the addition of liraglutide treatment to the ischaemia-reperfusion group (p < .05). First of all, we would like to say that liraglutide treatment is moderately preventive against I/R injury in testicular torsion. The anti-inflammatory, antioxidant and antiapoptotic properties of liraglutide are create a moderately protective effect as we show in this study.     

Protective influence of rosiglitazone against testicular ischaemia–reperfusion injury in rats

Testicular torsion is a urology urgent disease which causes testicular injury and potential sterility. In this study, we explored the protective influence of rosiglitazone on testicular ischaemia–reperfusion damage. There were 28 male Sprague Dawley rats in total, which were assigned randomly to four groups. Group A was blank control one; group B was testicular injury one; group C was rosiglitazone one; group D was rosiglitazone antagonist one. The testicles were counter‐rotated after 2 hr and then underwent orchiectomy 24 hr later. We found that testicular tissue structure of rats was seriously damaged in groups B and D. However, group C had better testicular architecture. Similar findings were also shown for lipid peroxidation by evaluating the MDA activity (p < .05). Unlike group B or group D, the levels of inflammation by evaluating the MPO activity, the levels of TNF‐a, IL‐1 and IL‐6 and the expressions of ICAM‐1 were prominently lower in group C (p < .05) as well. So our researches demonstrated that rosiglitazone significantly decreased the amount of responsive oxygen radical and regulated inflammatory responses. Rosiglitazone had a protective influence against testicular ischaemia–reperfusion injury in rats and possibly depended on its anti‐inflammatory and antioxidant traits.     

Effects of montelukast and zileuton on testicular torsion/detorsion injury in rats

The aim of this study was to evaluate and compare the effects of 5‐lipoxygenase enzyme (5‐LO) inhibitor zileuton and cysteinyl leukotriene receptor (CysLT1R) antagonist montelukast in testicular torsion/detorsion (T/D) injury model in rats. Rats were anaesthetised with 75 mg kg^?1 ketamine hydrochloride and 8 mg kg^?1 xylazine intraperitoneal before the operation. Torsion was created by rotating the right testis 720° clockwise and maintained by fixing the testis. The rats were treated with CysLT1R antagonist montelukast (10 mg kg^?1; i.p.), 5‐LO inhibitor zileuton (3 mg kg^?1; i.p.), and vehicle, at 30 min prior detorsion. After 1 h of torsion, the testis was counter‐rotated to the natural position and replaced into the scrotum. Malondialdehyde (MDA) level was measured in testicular tissue after 3 h of reperfusion. Histological examination was performed after 24 h of reperfusion. T/D caused a significant increase in MDA level and histopathological injury in testes. Montelukast and zileuton treatments prevented the T/D‐induced augmentation in MDA levels. Only zileuton treatment significantly reduced the T/D‐induced histopathological injury. In this study, we demonstrated for the first time that zileuton had protective effects on testicular T/D injury. We have also found that zileuton is more effective than montelukast on histopathological injury.     

Predictive value of ischaemia-modified albumin in spermatogenesis in an experimental testicular torsion model

Our aim was to measure the ability of ischaemia-modified albumin (IMA) to predict testicular histopathological damage in the testes of rats with short- and long-term ischaemia using experimental testicular torsion and subsequent reperfusion via detorsion.21 Wistar Albino rats were randomized into three groups. The sham group was subjected to a mid-scrotal incision only. The 4- and 8-hr T/D (Torsion/Detorsion) groups were subjected to left testicular torsion by twisting the testes by 720 degrees counterclockwise. 2 cc venous blood samples were taken from the sham group after the mid-scrotal incision, and from the 4- and 8-hr T/D groups after 4 and 8 hr respectively. After that, the 4- and 8-hr T/D groups were subjected to detorsion. Two days later, orchiectomy was performed. Ischaemia-modified albumin levels were significantly different among the groups at 48 hr prior to orchiectomy (reperfusion; p = .003). Based on the results of the paired comparisons, it was found that IMA levels of the sham group were significantly higher than those of the 4- and 8-hr T/D groups (p = .002 and .009 respectively). Our study has showed that IMA may be used to predict ischaemia/reperfusion injury, which is another complication that may occur following detorsion in testicular torsion.     

Effects of selenium on ischaemia–reperfusion injury in a rat testis model

Selenium is shown to have beneficial effects on ischaemia–reperfusion (IR) injury. Our aim was to assess the effects of selenium on IR‐induced testicular damage in terms of biochemical and histopathological evaluation. A total of 32 rats were randomised into four groups: control, IR, IR + selenium (IR + S) and S. Detorsion was applied after 3 h of torsion. Testicular tissue superoxide dismutase (SOD), glutathione (GSH), malondialdehyde (MDA), total antioxidant capacity (TAC) and DNA fragmentation levels were determined. Testicular tissue samples were examined by histopathological examination and terminal deoxynucleotidyl transferase dUTP nick end‐labelling staining. The control, IR and IR + S groups had higher SOD values compared with the S group; SOD levels of the control and IR + S groups were higher than those of the IR group (P < 0.05). Further, MDA levels of the IR group were higher than those in the other three groups (P < 0.05). The IR group revealed lower TAC levels than the three groups (P < 0.05 for all). GSH levels of the IR group were significantly lower than those in the other three groups (P < 0.05 for all). In contrast, GSH levels of the IR + S group increased compared with those of the S group. The IR group had more DNA fragmentation than the control and S groups (P < 0.05). It is concluded that selenium possibly reduces oxidative stress and apoptosis caused by testicular IR injury in rats. The testicular protective effect of selenium appears to be mediated through its anti‐apoptotic and antioxidative effects. However, selenium does not affect DNA fragmentation.     

Diffusion tensor imaging in evaluating testicular injury after unilateral testicular torsion and detorsion in rat model: A preliminary study

Diffusion tensor imaging (DTI) is a functional magnetic resonance sequence based on the movement of water molecules. This study attempted to investigate the feasibility of DTI in evaluating testicular injury after testicular torsion and detorsion. Seventy-two rats were randomly divided into the sham group, torsion group and detorsion group. The left testis in the sham group was brought out through a scrotal incision for 1 hr, and that of the torsion group was twisted 720^o clockwise for 1 hr and fixed to the scrotum, while the detorsion group was restored after being twisted 720° for 1 hr. Rats were further divided into four subgroups according to the set time, then performed DTI and histology analysis. The mean diffusion of the torsion and detorsion groups increased within 24 hr (p <.01), while it in the detorsion-1-week-group was lower than that in the detorsion-24-hr-group (p <.05). The fraction anisotropy of both experimental groups decreased in the acute phase (p <.01), while that of the detorsion-1-week-group increased (p <.01). Cosentino score in both experimental groups showed an increasing trend (p <.05). Besides, the spermatogenic ability of the detorsion-1-week-group decreased (p <.05). In conclusion, DTI was able to evaluate the injury after testicular torsion and detorsion.     

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.     

Dose-dependent protective effect of sildenafil citrate on testicular injury after torsion/detorsion in rats

This experiment was designed to investigate the effect of sildenafil citrate on testicular injury after unilateral testicular torsion/detorsion (T/D). Thirty-seven adult male Wistar albino rats were divided into four groups: sham operated group (group 1), T/D+ saline (group 2), T/D+ 0.7 mg sildenafil citrate (group 3) and T/D+ 1.4 mg sildenafil citrate (group 4). Testicular torsion was created by rotating the right testis 720° in a clockwise direction for 2 h in other groups, except for group 1, which was served as sham group. The level of GSH (P < 0.05) in the testis in the group 2 were significantly lower (P < 0.05) and the levels of MDA and NO (P < 0.01 for both) in the testis were significantly higher when compared with those of the group 1. Administration of low dose sildenafil citrate prevented the increases in MDA and NO levels and decreases in GSH values induced by testicular torsion. However, administration of high dose sildenafil citrate did not have any effect on these testicular tissue parameters (P > 0.05). Also, mean values of seminiferous tubules diameters, germinal cell layer thicknesses and mean testicular biopsy score were significantly better in group 3 than groups 2 and 4. These results suggest that T/D injury occurred in testis after unilateral testicular T/D and that administration of low dose sildenafil citrate before detorsion prevents ischemia/reperfusion cellular damage in testicular torsion. Sildenafil citrate probably acts through reduction of reactive oxygen species and support antioxidant enzyme systems.     

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° 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° 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.     

Protective effects of sumatriptan on ischaemia/reperfusion injury following torsion/detorsion in ipsilateral and contralateral testes of rat

This study was planned to evaluate the effects of sumatriptan, 5‐HT1B/1D receptors agonist, on ischaemia/reperfusion injury in bilateral testes after unilateral testicular torsion/detorsion in rats. Male Wistar rats (n = 42) were allocated into a sham‐operated group, a control group and treatment groups which were injected sumatriptan (0.1, 0.3 and 1 mg/kg), GR‐127935 (0.01 mg/kg)—5‐HT1B/1D receptors antagonist—and sumatriptan (0.1 mg/kg) + GR‐127935 (0.01 mg/kg). Torsion was induced for 1 hr by rotating right testis 720⁰ in the clockwise direction, and after 7 days of detorsion, bilateral orchiectomy was conducted. While the level of TNF‐α rose in testicular tissue after inducing torsion/detorsion, sumatriptan injection notably lowered TNF‐α level in ipsilateral (torted) and contralateral (nontorted) testes (p < 0.001). Moreover, after inducing testicular torsion/detorsion, SOD activity was decreased, whereas administration of sumatriptan significantly increased SOD activity in bilateral testes (p < 0.001). After induction of torsion/detorsion, macroscopic and histological analyses also showed severe damages which were improved by sumatriptan injection. Interestingly, co‐administration of sumatriptan with GR‐127935 reversed the beneficial impacts of sumatriptan on macroscopic appearance, microscopic pattern and biochemical markers. It is concluded that sumatriptan presumably via stimulation of 5‐HT_1B/1D receptors decreased inflammation, oxidative stress and deteriorations induced by ischaemia/reperfusion injury following testicular torsion/detorsion.     

Investigation of the protective role of chrysin within the framework of oxidative and inflammatory markers in experimental testicular ischaemia/reperfusion injury in rats

This study was performed to evaluate the effect of chrysin on testicular torsion and detorsion damage in rats in terms of biochemistry, histopathology and immunohistochemistry. The study was performed on Wistar albino rats between 250 g and 300 g. A total of 40 rats were used. Five groups were created with eight rats in each group. Group 1 was the control group, and no torsion procedure was performed. In Group 2, 2 hr of torsion and 2 hr of detorsion were applied. In Group 3, 2 hr of torsion and 24 hr of detorsion were applied. In Group 4, 2 hr of torsion, 2 hr of detorsion and 50 mg/kg intraperitoneal chrysin were applied. In Group 5, 2 hr of torsion, 24 hr of detorsion and 50 mg/kg of chrysin were applied. In the torsion/detorsion groups, the study determined decreases in glutathione and testosterone levels, increases in tumour necrosis factor-α, interleukin-4, interleukin-6 and interleukin-10 levels, and increases in expression levels of caspase-3 and caspase-8. Chrysin application reduced malondialdehyde, tumour necrosis factor-α, caspase-3 and caspase-8 expression levels. We can say that chrysin can be used to reduce damage in cases of testicular ischaemia/reperfusion. For more reliable results, further clinical trials are recommended.     

Protective effect of edaravone,a free‐radical scavenger,on ischaemia‐reperfusion injury in the rat testis

OBJECTIVE

To investigate the effect of edaravone, a radical scavenger, on ischaemia‐reperfusion (I‐R) injury in the testes.

MATERIALS AND METHODS

Eight‐week‐old male Sprague‐Dawley rats were allocated to one of four groups: a no‐drug group subjected to induction of 30‐min of ischaemia and 60‐min reperfusion; two drug groups administered edaravone at 1 or 10 mg/kg intraperitoneal and then subjected to 30‐min ischaemia and 60‐min reperfusion; and a sham‐operated control group administered edaravone at 10 mg/kg intraperitoneal. To induce testicular I‐R, the right testis was exposed outside of the body and the testicular artery was clamped with a small clip for 30 min. Blood flow and nitric oxide (NO) release were monitored in real time simultaneously with a laser Doppler flowmeter and an NO‐selective electrode, respectively. After death the tissue levels of NO₂‐NO₃ (a marker of NO production), malondialdehyde (a marker of lipid peroxidation), 8‐hydroxydeoxyguanosine (a marker of oxidative DNA damage), myeloperoxidase (a marker of neutrophil infiltration), and heat‐shock protein 70 (HSP 70) and its mRNA were measured. The testicular tissue was also analysed histologically.

RESULTS

Clamping the testicular artery resulted in a decrease of blood flow to 0–5% of the basal level measured before clamping. NO release was increased during clamping and gradually recovered to the basal level on removing the clip. Interestingly, the peak of NO release in rats of the no‐drug group occurred at the start of reperfusion, while that in the high‐dose drug group occurred several minutes later. The levels of NO₂‐NO₃, malondialdehyde, 8‐hydroxydeoxyguanosine, myeloperoxidase and HSP 70 and its mRNA, and histological variables, were significantly greater in the no‐drug I‐R group than in the control, and these variables were ameliorated by treatment with edaravone.

CONCLUSION

These results indicate that edaravone reduces the oxidative stress and prevents the testicular damage induced by I‐R.     

The effects of pirfenidone on ischaemia–reperfusion injury in testicular torsion-induced rat model

The aim of this study was to analyse the effect of pirfenidone against ischaemia–reperfusion injury occurring after detorsion in rats with induced testicular torsion model. Group 1 was assigned as the control group. Group 2 first had testis torsion performed, and then, testicular detorsion was performed. Group 3 had similar procedures to the rats in Group 2. Rats in Group 3 additionally had 325 mg/kg pirfenidone administered immediately after ischaemia. The blood samples were analysed spectrophotometrically. To determine the intensity of tissue injury, haemorrhage, oedema and congestion levels were evaluated with direct microscopic investigation of testis. Seminiferous tubule architecture, spermatogenesis processes and germ cell maturation were graded by Johnsen and Cosentino scoring systems. In Group 3, superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities increased compared with Group 2 (p:.03 and p:.049 respectively). Additionally, the mean malondialdehyde (MDA) value was higher in Group 2 compared with the other groups (p:.001). Histopathological investigation of rats in Group 3 identified positive changes in haemorrhage, oedema and congestion levels compared with Group 2 (p:.031, p:.048, p:.044 respectively). Similarly, Johnsen and Cosentino scores were positively affected in Group 3 (p:.033, p:.032 respectively). Pirfenidone is protective against testicular oxidative damage.     

一氧化氮在一侧睾丸扭转对侧睾丸损伤中的作用

目的 研究总抗氧化能力(T-AOC)和一氧化氮(NO)在一侧睾丸扭转对侧睾丸损伤中的作用。方法 SD雄性大白鼠建立左侧睾丸扭转模型，于扭转后6h再分为扭转睾丸复位及切除组，分别于术后1h、1d、1周、2周和4周处死4—5只，取出睾丸用于一氧化氮合酶(NOS)活性、NO、T-AOC及细胞凋亡的检查。结果 UTT复位后对侧睾丸组织NOS活性、NO含量明显升高，T—AOC显著降低。结论 NO过量产生及T-AOC的下降是UTT对侧睾丸损伤的重要原因。     

The protective effect of darbepoetin alfa on experimental testicular torsion and detorsion injury

AIM: Testicular torsion is a serious urological emergency, usually involving newborns, children, and adolescents which can lead to subfertility and infertility. Prevention of testicular damage caused by torsion is still a clinical and experimental problem. So far many chemicals and drugs have been investigated for decreasing ischemia/reperfusion (I/R) injury in experimental animals. The possible protective effect of darbepoetin alfa, a novel erythropoietic protein, on testicular tissue after I/R injury was examined in this study. METHODS: Thirty rats were divided into three groups: sham operation, torsion/detorsion, and torsion/detorsion plus darbepoetin alfa groups. After torsion (2 hours) and detorsion (4 hours), bilateral orchiectomy was performed. Malondialdehyde, nitric oxide and glutathione levels were determined in testicular tissue. RESULTS: Administration of darbepoetin alfa caused a decrease of malondialdehyde and nitric oxide levels and an increase in glutathione levels compared with the torsion/detorsion group. In addition, histological injury scores were significantly decreased in the treatment group more than the torsion/detorsion group. CONCLUSION: The results suggest that darbepoetin alfa may be a potential protective agent for preventing testicular injury caused by testis torsion.     

Protective effects of quercetin on testicular torsion/detorsion-induced ischaemia-reperfusion injury in rats

The aim of this study was to investigate the protective effect of quercetin (QE) on testicular torsion/detorsion-induced ischaemia-reperfusion (I/R) injury. A total of 24 male Wistar albino rats were divided into three groups: control, I/R and I/R treated with QE; each group contain eight animals. Testicular torsion was created by rotating the left testis 720° in a clockwise direction. The ischaemia period was 5 h and orchiectomy was performed after 5 h of detorsion. QE (15 mg kg(-1) , i.p.) was administered only once, 40 min prior to detorsion. Left orchiectomy was performed in all I/R groups. To date, no histopathological changes on testicular torsion/detorsion-induced I/R injury in rats by QE treatment have been reported. Spermatogenesis and mean seminiferous tubule diameter were significantly decreased in I/R groups were compared with the control group. Furthermore, QE treated animals showed an improved histological appearance in I/R group. Our data indicate a significant reduction in the activity of TUNEL, endothelial nitric oxide synthase and a rise in the expression of testosterone in testes tissue of I/R treated with QE therapy. We believe that further preclinical research into the utility of QE may indicate its usefulness as a potential treatment on testes injury after I/R in rats.