The role of nitric oxide in testicular ischemia-reperfusion injury

PURPOSE: This study was designed to determine the role of nitric oxide (NO) in the ischemia-reperfusion (I/R) injury process in testes. METHODS: Fifty prepubertal male rats were divided into 5 groups each containing 10 rats. After 4-hour torsion and 4-hour detorsion, bilateral orchiectomies were performed for measurement of tissue malondialdehyde (MDA) level and histopathologic examination. The results were compared statistically. The groups were labeled as group 1, basal values of biochemical parameters in testes; group 2 (control group), torsion plus detorsion; group 3, torsion plus N-monomethyl-L-arginine (L-NMMA) plus detorsion; group 4, torsion plus L-arginine plus detorsion; group 5, sham operation. RESULTS: The highest MDA values were determined in the L-arginin group in ipsilateral testes. Group 3 and group 4 were statistically different from control group. Histological examination showed that specimens from group 4 had a significantly (P < .05) greater histological injury than group 3, and contralateral testes showed normal testicular architecture in all groups. CONCLUSIONS: These results suggest that NO plays an important role in damaging the testis with I/R. Although inhibition of NO synthesis with L-NMMA significantly improves I/R injury in testes, enhancing NO production by providing excess of L-arginine increases such damage. In the early periods of detorsion, there is no damage to contralateral testes after unilateral testicular torsion.     

Dehydroepiandrosterone treatment attenuates reperfusion injury after testicular torsion and detorsion in rats

Purpose

We aimed to evaluate the effects of dehydroepiandrosterone (DHEA) on antioxidant enzyme activities, lipid peroxidation, and histopathologic changes in both testes after unilateral testicular torsion and detorsion.

Methods

Twenty-four adult male Sprague-Dawley rats were randomly divided into 4 groups (n = 6 for each group): sham operation, torsion/detorsion (T/D), T/D + vehicle, and T/D + DHEA. Three hours before detorsion, 50 mg/kg DHEA was given intraperitoneally to the T/D + DHEA group. Testicular ischemia was achieved by twisting the left testis 720° clockwise for 3 hours, and reperfusion was allowed for 24 hours after detorsion. In all groups, bilateral orchiectomies to determine the testicular tissue catalase (CAT) and superoxide dismutase activities and malondialdehyde (MDA) levels and histopathologic examination were performed.

Results

Compared with those from the sham group, CAT activities in the ipsilateral testis obtained from the T/D group were significantly lower and MDA levels were significantly higher (P < .05 for all).Administration of DHEA prevented increases in MDA levels and decreases in CAT and superoxide dismutase activities when compared to the T/D group. Specimens from the T/D and the T/D + vehicle groups had a significantly greater histologic injury than the specimens from the sham and the T/D + DHEA groups had (P < .01 for both).

Conclusions

The results suggest that DHEA may be a protective agent for preventing biochemical and histopathologic changes related to oxidative stress in testicular injury caused by testis torsion.     

Protective role of methylprednisolone and heparin in ischaemic‐reperfusion injury of the rat testicle

This study evaluated the therapeutic efficacy of heparin and methylprednisolone in the treatment of ischaemic reperfusion (IR) injury of the testis. Twenty‐four male Sprague‐Dawley rats were allocated equally into three groups of eight animals each. The left testes were rotated 720° for 2 h in the rats in the torsion–detorsion group. Rats in the treatment groups underwent the same surgical procedure as the torsion–detorsion group but were also given methylprednisolone (group II) or heparin (group III) by an intraperitoneal route 30 min prior to detorsion. Left orchiectomy was performed in all rats from each experimental animal at 2 h after detorsion, and the tissue was harvested for the measurement of malondialdehyde (MDA), protein carbonyl (PC) and nitric oxide (NO) and the endogenous antioxidant enzymes, superoxide dismutase (SOD), glutathione peroxidase (GSH‐Px) and catalase. Additional tissue was evaluated using histopathological and immunohistochemical changes. PC and MDA levels were significantly reduced in the treated groups compared to the control group. There was no statistically significant difference in NO level or SOD, GSH‐Px and catalase activity among the treatment groups. Histopathological and immunohistochemical findings supported biochemical changes. It is concluded that pre‐treatment with methylprednisolone or heparin protects the testis in ischaemic reperfusion injury caused by testicular torsion–detorsion.     

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.     

The effects of nitric oxide on the expression of cell adhesion molecules (ICAM-1, UEA-1, and tenascin) in rats with unilateral testicular torsion

Background/Purpose:

The aim of this study was to determine the effects of nitric oxide (NO) on the expression of adhesion molecules in the early course of testicular I-R injury in rats.

Methods:

Forty male Sprague-Dawley rats were separated into 4 groups, each containing 10 rats. A sham operation was performed in group 1 (control). In group 2 (I-R), after 6 hours of unilateral testicular torsion, 1-hour detorsion of the testis was performed. In group 3 (I-R/l-NAME), after performing the same surgical procedures as in group II, l-NAME was given for 30 minutes. In group 4 (I-R/Mol), after performing the same surgical procedure (torsion and detorsion) as in group II, molsidomine, an NO donor, was infused for 30 minutes. Then, ipsilateral orchiectomies were performed to measure the tissue levels of malondialdehyde (MDA) and NO and to make histologic examination.

Results:

MDA values and the testicular injury score decreased and NO values increased in the I-R/Mol-treated group compared with other experimental groups. The tenascin expression in the interstitial space and basement membrane of the tubuli seminiferi were milder in the I-R/Mol group compared with that of the I-R and the I-R/l-NAME. The acrosomes of the spermatids in I-R and I-R/l-NAME groups were stained mildly by lectin. In the I-R and I-R/l-NAME groups, the interstitial spaces, basement membrane of the tubuli seminiferi, and sertoli and germinal cells in testicular tissue were stained intensely by ICAM-1.

Conclusions:

The expression of adhesion molecules such as tenascin, lectin, and ICAM-1 in the torted testicular tissue may be a pathophysiologic sign of inflammation. NO regulates adhesion molecules expression.     

Beneficial effects of melatonin compared with allopurinol in experimental testicular torsion

Background/purpose

Several antioxidant agents such as allopurinol have been used to prevent ischemia-reperfusion (I/R) injury-induced tissue damage after experimental testicular torsion so far. The current study was designed to determine the effect of melatonin, which is a potent antioxidant agent, in preventing testicular damage following torsion.

Methods

Sixty prepubertal male Wistar-Albino rats were divided into 5 groups: control (C), torsion (T), torsion plus detorsion (TD), torsion plus allopurinol (200 mg/kg) plus detorsion (A), and torsion plus melatonin (50 mg/kg) plus detorsion (M). Left testes were rotated 720° for 6 hours. The torsed testes were detorsed. Detorsion time was 6 hours. In all groups, left orchiectomies were performed to determine the tissue levels of malondialdehyde (MDA) and histopathologic changes. Blood samples were taken to measure serum creatine phosphokinase (CPK) levels. The results were analyzed statistically.

Results

Serum CPK levels of groups A and M were found to be significantly lower than groups T and TD (P < .05). Tissue MDA levels in group M were statistically different from groups T and TD (P < .05). However, in groups A and T, MDA levels were similar (P > .05). The highest histologic grade was determined in group TD (3.8 ± 0.5). Histologic grade of group M was significantly lower than group TD (P < .001), but there was no histologic difference between testes of groups A and TD (P > .05).

Conclusions

These results have shown that melatonin treatment prevents I/R injury both biochemically and histopathologically, whereas allopurinol treatment prevents it only biochemically in experimental testicular torsion. Melatonin is a potent antioxidant agent more effective than allopurinol in preventing testicular I/R injury.     

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.     

Propofol attenuates reperfusion injury after testicular torsion and detorsion

Propofol, which is widely used as an intravenous anesthetic, has been shown to have an antioxidant activity on several tissues. This study was designed to investigate the prevention of reperfusion injury with propofol after testicular torsion. Five groups of rats (seven in each group) were used. Animals in the control group (group I) did not received any treatment, while animals in the sham group (group II) underwent scrotal incision and testicular fixation only. After 2 h of 720° left testicular torsion in groups III, IV and V, subsequent detorsion was done for 2 h in groups IV and V. Propofol (50 mg/kg) was injected transperitoneally 30 min prior to detorsion in group V. Both testicles in all rats were retrieved and tissue malondialdeyhde (MDA) level, which is a measure of the amount of free oxygen radicals, and enzymatic activity of xanthine oxidase (XO), which converts hypoxanthine to xanthine and uric acid were studied. In addition, tissue catalase (CAT) and glutathione peroxidase (GSH-Px) activities, which are endogenous scavenger enzymes, protecting tissues against free radicals, were studied. Additionally, histological evaluations were performed after hematoxylin and eosin staining. Testicular MDA levels, and XO and CAT activities were higher in the torsion group compared to sham control group (P<0.05). Detorsion caused a further increase in MDA levels, contrasting with a decrease in the levels of XO activity, while CAT activity was not changed. Pretreatment with propofol prevented a further increase in MDA levels and significantly decreased CAT activity following detorsion. GSH-Px activities were not effected either by torsion/detorsion or propofol pretreatment. Histologically, torsion caused some separation between germinal cells in the seminiferous tubules, which became much more prominent in the detorsion group and attenuated with propofol pretreatment. There was no significant change in any of the abovementioned enzymatic activities nor were there histopathological changes in the contralateral testicle in any groups. It is concluded that biochemically and histologically reperfusion injury occurs in the ipsilateral testis following detorsion up to 2 h. Preference of propofol for anaesthesia during the detorsion procedure may attenuate such reperfusion injury.     

黄芪注射液对大鼠扭转复位后睾丸组织的保护作用

目的:探讨黄芪注射液对雄性Wistar大鼠扭转复位后睾丸的保护作用。方法:30只大鼠随机分为假手术组(A组)、睾丸扭转复位组(B组)、黄芪注射液治疗组(C组),每组10只,Turner法建立单侧睾丸扭转复位模型,原位缺口末端标记法检测各组睾丸组织中生殖细胞凋亡,化学比色法测定超氧化物歧化酶(SOD)和丙二醛(MDA)含量。结果:黄芪注射液治疗组与睾丸扭转复位组比较,SOD含量明显升高,MDA含量明显降低,生精细胞凋亡指数明显降低。睾丸扭转复位组、黄芪注射液治疗组与假手术对照组比较,SOD含量明显降低,MDA含量明显升高,生精细胞凋亡指数明显升高。结论:黄芪注射液可减少大鼠睾丸扭转复位后睾丸组织的双侧睾丸生殖细胞凋亡,对扭转复位后睾丸生殖细胞有保护作用。其机理可能与提高抗氧化酶活性及减少氧自由基的产生从而减轻大鼠睾丸扭转复位后的缺血再灌注损伤有关。     

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

Objectives  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. Methods  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. Results  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. Conclusion  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.     

Pentoxifylline attenuates reperfusion injury in testicular torsion

OBJECTIVES: An experimental study was designed to evaluate the effects of pentoxifylline (Ptx) on lipid peroxidation, and histopathology in both testes after unilateral testicular torsion and detorsion. MATERIALS AND METHODS: Forty adult male albino Wistar rats were randomly divided into 4 groups of sham operation, sham operation with Ptx, torsion and detorsion, torsion and detorsion with Ptx. After intraperitoneal administration of Ptx at a dose of 50 mg/kg 15 min before torsion; right testes of the rats underwent 30 min of torsion and 30 min of detorsion. Malondialdehyde (MDA) levels were assayed and histopathological changes were evaluated in both testes of all groups. RESULTS: Unilateral testicular torsion and detorsion caused an increase in the MDA levels of both testes. Histopathological evaluation showed interstitial hemorrhage on the ipsilateral side. Pentoxifylline decreased MDA levels on both side, and attenuated interstitial injury on the ipsilateral side. CONCLUSIONS: The results of this study suggest that pentoxifylline treatment attenuates reperfusion damage on both side, possibly with its effects on blood flow and neutrophils. However, further studies are necessary to evaluate the effects of pentoxifylline on testicular torsion.     

The preventive effects of thiopental and propofol on testicular ischemia-reperfusion injury

BACKGROUND: Testicular torsion is a urological emergency that requires immediate surgical intervention to prevent testicular damage. The aim of the study was to investigate the preventive effects of thiopental and propofol as anesthetics on testicular ischemia-reperfusion injury. METHODS: Forty male Wistar Albino rats were randomly assigned to four groups of 10 rats each. During 5 h, anesthesia was induced and maintained with thiopental in groups 1 and 2 and with propofol in groups 3 and 4. Groups 2 and 4 received left testicular ischemia (torsion) during 1 h and reperfusion (detorsion) during 4 h. Groups 1 and 3 (control groups) had no testicular torsion and detorsion. At the end of 5 h, animals were killed and both ipsilateral and contralateral testes were removed for histopathologic examination and measurement of tissue MDA (malondialdehyde) and NO (nitric oxide) levels. RESULTS: In the contralateral testes of all the groups, MDA and NO measurements were not different from ipsilateral testes of the control groups. Between the groups 1 and 3, there were no differences in MDA and NO levels. Although torsion/detorsion of testes in group 4 caused significantly increased levels of tissue MDA and NO values compared with group 3, ischemia-reperfusion in group 2 caused a further increase in these levels compared with group 4. The ipsilateral testes in the control groups did not show any morphological changes. Testicular torsion/detorsion in rats with thiopental anesthesia (group 2) caused significantly greater histopathologic injury levels than rats with propofol anesthesia (group 4). CONCLUSION: Our results suggest that propofol as an anesthetic agent may prevent testicular damage by scavenging reactive oxygen and nitrogen species and inhibiting lipid peroxidation in an animal model of testicular torsion and detorsion.     

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.     

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.     

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 effects of the hydroalcoholic extract of Fumaria parviflora on testicular injury induced by torsion/detorsion in adult rats

This study was designed to determine the effects of daily oral administration (250 mg/kg) of the hydroalcoholic extract of Fumaria parviflora (FP) for 14 days on the sperm parameters, oxidative stress parameters, serum testosterone levels, expression of Bax and Bcl‐2 genes, and apoptosis index of germ cells after testicular torsion–detorsion (ischaemia–reperfusion, IR) injury model in rats. Twenty‐eight adult male Wistar rats were divided randomly into four groups of seven each: sham operation, torsion–detorsion (TD), TD plus the hydroalcoholic extract FP (TDFP) and only FP without TD application (FP). Testicular torsion was created by rotating the left testis 720° in a counterclockwise direction; then, after 4 hr, detorsion was performed. The Johnson's score, mean seminiferous tubule diameter (MSTD) and height (thickness) of seminiferous tubule epithelium (HST) were significantly increased in TDFP and FP groups as compared to TD group. The gene expression of Bcl‐2, level of serum testosterone hormone and antioxidant parameters—GPx and SOD—were significantly higher in TDFP and FP groups than TD group. The index of apoptosis, the gene expression of Bax and the level of MDA were significantly higher in TD group than TDFP and FP groups. Therefore, F. parviflora could decrease oxidative stress induced by testicular torsion–detorsion.     

Effect of insulin-like growth factor-1 on apoptosis of rat testicular germ cells induced by testicular torsion

OBJECTIVE: To investigate the possible protective role of insulin-like growth factor-1 (IGF-1, reported to have a protective effect in experimental models of hypoxic ischaemia), and the involvement of apoptotic cell death in a model of torsion/detorsion of the rat testis. MATERIALS AND METHODS: Adult male Wistar rats were divided into five groups of five rats each. Group 1 underwent a sham operation as a control; in group 2 the testis was twisted and in group 3 then untwisted; in group 4 IGF-1 was injected subcutaneously just before bilateral torsion, and then the right testis removed after 4 h and the left after 24 h; in group 5, IGF-1 was injected immediately after bilateral detorsion and then the testes removed as in group 4. Both testicles were examined histologically, with apoptosis detected using the in situ DNA fragmentation (TUNEL) system, with combined enzymology and immunohistochemistry techniques. RESULTS: In groups 2 (torsion) and 3 (detorsion), light microscopy of the testis showed some degenerative changes in the germ cells. Compared to group 1, apoptosis was more significant in group 3 than in the other groups. Group 4 (torsion/IGF-1) had a similar number of apoptotic germ cells as in group 2 (torsion) after 24 h, but fewer than the same group after 4 h. In group 5 (detorsion/IGF-1), apoptosis was reduced by IGF-1 significantly more than in group 3 (P < 0.05). Apoptosis was significantly less in spermatids in group 5 than in group 3 (P < 0.05). CONCLUSIONS: IGF-1 seems to lower the levels of germ cell apoptosis, which may be important for protecting the testes from torsion/detorsion injury. Further clinical studies are needed to evaluate this protective effect in testicular torsion/detorsion.     

单侧睾丸扭转对生殖细胞凋亡及黄芪保护作用的实验研究

目的观察大鼠单侧睾丸扭转／复位后患侧和对侧睾丸生精细胞凋亡情况,探讨单侧睾丸扭转／复位后生殖能力下降的机制以及黄芪注射液对其再灌注损伤的保护作用。方法将40只健康雄性Wistar大鼠分为4组,分别为假手术对照组（A组）,睾丸扭转／复位组（B组）,睾丸扭转／复位＋单次腹腔内注射黄芪注射液组（C组）及扭转／复位十连续腹腔内注射黄芪注射液组（D组）,每组10只。按Turner法建立睾丸扭转／复位模型,所有大鼠均在同等条件下喂养至术后7d处死,切取双侧睾丸后检测凋亡指数。结果扭转侧睾丸生殖细胞凋亡指数（AI）A组（5．82±1．21）与B组（36．18±8．40）、C组（20．39±3．57）、D组（11．61±5．12）相比差异有显著性（P〈0．05）,B组明显高于C组及D组（P〈0．05）,C组与D组相比差异有显著性（P〈0．05）;B组对侧睾丸（12．95±3．06）与C组（9．45±1．71）、D组（7．56±1．06）两组对侧睾丸AI相比差异有显著性（P〈0．05）,C、D两组对侧睾丸AI差异有显著性（P〈0．05）。结论单侧睾丸扭转可致患侧和对侧睾丸生精细胞凋亡明显增加,黄芪注射液可明显减少双侧睾丸生殖细胞凋亡,连续应用黄芪注射液优于单次应用。     

Reperfusion injury after detorsion of unilateral testicular torsion

Summary Reperfusion injury has been well documented in organs other than testis. An experimental study was conducted to investigate reperfusion injury in testes via the biochemical changes after unilateral testicular torsion and detorsion. As unilateral testicular torsion and varicocele have been shown to affect contralateral testicular blood flow, reperfusion injury was studied in both testes. Given that testicular blood flow does not return after 720° testicular torsion lasting more than 3 h, the present study was conducted after 1 and 2 h of 720° torsion. Adult male albino rats were divided into seven groups each containing ten rats. One group served to determine the basal values of biochemical parameters, two groups were subjected to 1 and 2 h of unilateral testicular torsion respectively, two groups were subjected to detorsion following 1 and 2 h of torison respectively, and two groups underwent sham operations as a control. Levels of lactic acid, hypoxanthine and lipid peroxidation products were determined in testicular tissues. Values of these three parameters obtained from the sham operation control groups did not differ significantly from basal values (P>0.05). All three parameters were increased significantly in both ipsilateral and contralateral testes after unilateral testicular torsion when compared with basal values (P<0.01 and P<0.05, respectively). Detorsion caused significant changes in lipid peroxidation products levels in ipsilateral but not in contralateral testes when compared with values obtained after torsion (P<0.01 and P>0.05, respectively). It is concluded that ipsilateral testicular torsion causes a decrease in perfusion not only in the ipsilateral but also in the contralateral testis. Additionally, detorsion following up to 2 h of 720° torsion causes reperfusion injury in ipsilateral but not in contralateral testis.     

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

Aim

Testicular 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 Methods

Thirty-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.

Results

Tissue 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).

Conclusion

We 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.