Protective effects of Fumaria parviflora L. on lead‐induced testicular toxicity in male rats

In recent years, the clinical importance of herbal drugs has received considerable attention in reducing free radical‐induced tissue injury. Oxidative stress has been proposed as a possible mechanism involved in lead toxicity that causes reproductive system failure in both human and animals. Fumaria parviflora L., a traditional herb, has been used to cure various ailments in Persian folk medicine. This study was carried out to investigate whether ethanolic extract of F. parviflora leaves could protect the male rats against lead‐induced testicular oxidative stress. Adult Wistar rats were treated with 0.1% lead acetate in drinking water with or without 200 mg kg day^?1 F. parviflora extract via gavage for 70 days. Lead acetate treatment resulted in significant reduction in testis weight, seminiferous tubules diameter, epididymal sperm count, serum testosterone level, testicular content of superoxide dismutase (SOD) and glutathione peroxidase (GPx). Moreover, significant elevation was observed in content of malondialdehyde (MDA) in lead‐treated rats. However, co‐administration of F. parviflora extract showed a significant increase in selected reproductive parameters in lead‐treated rats. The results indicated that ethanolic extract of F. parviflora leaves has a potential to restore the suppressed reproduction associated with lead exposure and prevented lead‐induced testicular toxicity in male Wistar rats.     

Sinapic acid reduces ischemia/reperfusion injury due to testicular torsion/detorsion in rats

This study aimed to investigate the protective effect of sinapic acid (SA) on biochemical and histopathological changes in an experimental testicular torsion-detorsion rat model. Twenty-four rats were randomised into four groups: sham group, ischemia/reperfusion (IR) group subjected to testicular torsion for 2 hr and then detorsion for 4 hr, and two groups treated with SA1 and SA2 (10 mg/kg and 20 mg/kg, by single intraperitoneal injection, 30 min before reperfusion). Serum testosterone, follicle-stimulating hormone (FSH), and luteinizing hormone (LH) were measured by an autoanalyzer, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), malondialdehyde (MDA), protein carbonyl (PC), and nitric oxide (NO) oxidative stress parameters by spectrophotometric methods, and tumour necrosis factor (TNF-α), interleukin-1 beta (IL-1β), and interleukin 6 (IL-6) parameters by the Elisa method. In addition, immunohistochemical and histopathological examinations were performed on testicular tissues. There was no significant difference between the groups in terms of serum testosterone, FSH and LH levels (p > .05). SA significantly reduced increased testicular damage, oxidative stress, inflammation, cell death and also restored decreased antioxidant enzyme activities (p < .05). Pre-treatment of rats with SA reduced testicular dysfunction and morphological changes IRI. SA's antioxidant, anti-inflammatory, and antiapoptotic properties were found to be protective against testicular IR.     

葡萄籽原花青素对小鼠睾丸扭转复位后生精功能的保护作用

目的:探讨葡萄籽原花青素(GSP)对小鼠睾丸扭转复位后生精功能的保护作用。方法:24只健康雄性昆明小白鼠(8周龄,25～27 g)随机分为3组:对照组、扭转组、治疗组,每组8只。扭转组及治疗组建立单侧睾丸扭转复位动物模型,治疗组于扭转复位前30 min腹腔注射GSP(50 mg/kg),术后采用腹腔注射方式连续给药3 d,每天1次,每次50 mg/kg。扭转组方法同治疗组,治疗同体积生理盐水。术后第4天取扭转侧睾丸,检测组织病理学参数和生精细胞凋亡指数(AI),并检测睾丸组织超氧化物歧化酶(SOD)和丙二醛(MDA)含量。对照组行假手术。结果:治疗组与扭转组相比,Johnsen评分上升[(7.38±0.92)分vs(5.00±1.85)分,P<0.05],生精小管直径略增大[(178.75±1.58)μm vs(176.50±1.60)μm,P>0.05],生精细胞层数增加[(5.75±0.71)层vs(3.75±1.03)层,P<0.05],生精细胞凋亡指数AI明显降低[(16.25±1.67)%vs(40.50±1.60)%,P<0.05)],SOD活性明显上升[(52.67±3.57)U/mg prot vs(29.04±4.46)U/mg prot,P<0.05],MDA含量明显下降[(2.91±0.04)nmol/mg prot vs(4.63±0.05)nmol/mg prot,P<0.05]。结论:GSP对小鼠睾丸扭转复位后生精功能损伤有明显的保护作用,其作用机制可能与其能清除氧自由基、抑制脂质过氧化、提高机体抗氧化能力有关。     

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.     

Protective effect of thymoquinone against testicular torsion induced oxidative injury

We aimed to determine the protective effects of thymoquinone (TQ), against ischaemia–reperfusion (I/R) injury in the testis tissue of rats. Twenty‐seven male Wistar albino rats were randomly divided into three equal groups as follows: Group I, sham group; Group II, torsion group; and Group III, torsion + thymoquinone group. The ischaemia period was 2 h, and orchiectomy was performed after 30 min of detorsion. Testis tissue sections were analysed with the terminal transferase mediated dUTP‐nick end labelling (TUNEL) assay to determine in situ apoptotic DNA fragmentation. Additionally, Caspase 3 and Bax proteins were analysed immunohistochemically. The superoxide dismutase (SOD), glutathione peroxidase (GSH‐Px) and malondialdehyde (MDA) activity levels in the testis tissue were also measured. The superoxide dismutase activity and malondialdehyde levels in the torsion group were significantly higher than those of the sham group (P < 0.05). Thymoquinone administration significantly reduced these levels. Torsion significantly increased active‐Caspase 3 and Bax expression, which was decreased by thymoquinone. The apoptotic index of the torsion group was significantly higher than that of the control group. However, thymoquinone significantly reduced the apoptotic index (P < 0.05). Our results indicate that thymoquinone plays a protective role in oxidative stress induced ischaemia–reperfusion in the testis tissue of rats.     

Fumaria parviflora regulates oxidative stress and apoptosis gene expression in the rat model of varicocele induction

Varicocele is one of the leading causes of male infertility in which oxidative stress induces DNA damages in spermatozoa of patients with varicocele. Recent studies indicated that the treatment with antioxidant agents has protective effects against the formation of reactive oxygen species (ROS). Our research aimed to evaluate the impact of Fumaria Parviflora (FP) on the varicocele-induced testicular injury. For this purpose, 32 adult male Wistar rats (n = 8 per group) were randomly assigned to four groups as follows: sham group, varicocele group, varicocele treatment group and the control treatment group. The experimental groups daily received FP (250 mg/kg) for 8 weeks. The induction of varicocele was conducted by partial occlusion on the left renal vein. The diameter of seminiferous tubules, Johnsen's score and the epithelium thickness improved in the treated-varicocele group as compared to the varicocele group. FP extract could increase the biochemical parameters including superoxide dismutase and glutathione peroxidase, and also decrease malondialdehyde level in the varicocele group. Furthermore, varicocele markedly increased both mRNA and intensity of Bax, while treatment with FP could alleviate them. We concluded that FP could alleviate varicocele, possibly by lowering oxidative stress and testicular damage.     

Co-administration of Salvia miltiorrhiza and verapamil inhibits detrimental effects of torsion/detorsion on testicular tissue in rats

Testicular torsion/detorsion is one of the important emergencies that requires fast surgical intervention. This study aimed to investigate the effects of Salvia miltiorrhiza hydroalcoholic extract combined with verapamil on testicular ischaemia/reperfusion damage in Wistar albino rats. All animals were distributed in 3 groups (n = 8), including the sham-operated group, torsion/detorsion (TD) group and torsion/detorsion + pretreatment with 200 mg/kg Salvia miltiorrhiza extract combined with 0.3 mg/kg verapamil (SMV) group. Oxidative stress biomarkers (MDA, GPx, CAT and TAC) both in plasma and testicular tissue, sperm parameters (motility, vitality, concentration and morphology) and histopathological parameters (MSTD, GECT, Johnson's score, Cosentino's score and testicular cell thickness) were assessed in all groups. Ischaemia/reperfusion significantly increased MDA and decreased GPx, CAT and TAC levels (p < .05). Pretreatment with SMV significantly increased GPx, CAT and TAC levels (p < .05). SMV group increased progressive sperm motility and vitality and reduced non-progressive motility of spermatozoon (p < .05). Testicular torsion significantly decreased all histopathological parameters compared to the sham group (p < .05). SMV pretreatment remarkably increased MSTD, GECT and Cosentino's score in comparison with the TD group (p < .05). A combination of Salvia miltiorrhiza with verapamil could reduce damages triggered by testicular torsion detorsion and improve sperm functionality parameters and oxidative stress defence systems.     

The effects of verapamil and heparin co-administration on sperm parameters and oxidative stress in prevention of testicular torsion/detorsion damage in rats

In this research, the impacts of combined administration of verapamil and heparin on testicular torsion damage were examined. In this experimental study, 30 sexually mature male Wistar albino rats were divided into five equal groups haphazardly (n = 6): Group 1 was the sham group. In group 2, a 2-hr testicular torsion was induced, and thereafter, detorsion was done. Rats in group 3 and group 4 experienced an identical surgical procedure like group 2, but verapamil and heparin were administered in 0.3 mg/kg and 800 IU/kg doses respectively, and in group 5, a combination of verapamil and heparin were administered. Intraperitoneal drug injection in all treatment groups was done 30 min before testicular detorsion. Testicular torsion significantly changed sperm parameters, oxidative stress biomarkers and Cosentino's histological score compared to the sham group (p < .05). All treatment groups reduced testicular damage by decreasing oxidative stress and improving sperm parameters, but heparin and co-administration of verapamil and heparin were significantly better than verapamil injection alone. However, heparin injected group was more effective than other treatment groups (p < .05). Overall, an anticoagulant like heparin is more effective than a calcium channel blocker such as verapamil, and it is more likely to reduce testicular torsion injuries.     

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.     

Protective role of Diospyros lotus on cisplatin‐induced changes in sperm characteristics,testicular damage and oxidative stress in rats

The aim of this study was to investigate the protective effect of Diospyros lotus (DL) on cisplatin (CP)‐induced testicular damage in male rats. Twenty‐eight male rats were randomly divided into four groups: group 1 – control, given isotonic saline solution; group 2 – CP 7 mg kg⁻¹ given intraperitoneally as single dose; group 3 – DL 1000 mg kg⁻¹ per day given orally for 10 days; group 4 – CP and DL given together at the same doses. CP caused a significant increase in thiobarbituric acid‐reactive substances (TBARS) level and a significant decrease in superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT) and glutathione (GSH) levels in rats testis tissues compared to the control group. CP caused a significant increase in lipid peroxidation in testis tissues compared to the control group, whereas DL led to a significant increase in SOD and GSH levels. However, there were no statistically significant changes in GPx and CAT levels. In addition, serum testosterone levels, sperm concentration and sperm motility were significantly decreased, but abnormal sperm rate and histological changes were increased with CP. However, these effects of CP on sperm parameters, histological changes and the tissue weights were eliminated by DL treatment. In conclusion, our study showed that the reproductive toxicity caused by CP may be prevented by DL treatment.     

Protective efficiency of Ashrasi date palm hydroalcoholic extract against diabetes‐induced testicular toxicity: A biochemical and stereological study

The present study was designed to investigate the protective effects of hydroalcoholic extract of Ashrasi date palm (ADP) on diabetes‐induced testicular injuries. Adult male rats were randomly allocated into five groups (n = 8 in each group): 1: control; 2: diabetic; 3: diabetic + 30 mg/kg of ADP extract; 4: diabetic + 90 mg/kg of ADP extract; and 5: diabetic + 270 mg/kg of ADP extract. Diabetes was induced by a single dose of streptozotocin (55 mg/kg) intraperitoneally. Testicular changes were assessed quantitatively using stereological method followed by measuring antioxidant enzymes including catalase, superoxide dismutase and glutathione peroxidase and serum testosterone level. Malondialdehyde (MDA) and Bcl‐2 expression were also evaluated in tissue samples. Diabetes resulted in significant deleterious alterations in the architecture of testicular tissue, suppressed antioxidant enzymes and testosterone levels and increased lipid peroxidation. The expression of Bcl‐2 was downregulated in diabetic testis and resulted in enhanced apoptosis. Eight weeks of ADP extract treatment especially at higher doses could markedly improve structural changes of testis and restore the antioxidant defence and testosterone levels in testicular tissue. In conclusion, this findings showed that ADP extract can play as a potent antioxidant and can attenuate the adverse effects of diabetes on male reproduction.     

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.     

Protective effects of sea cucumber (Holothuria atra) extract on testicular dysfunction induced by immune suppressant drugs in Wistar rats

The current study was aimed to evaluate the protective effect of Holothurian atra (HA) extract; naturally occurring marine resource, against methotrexate (MTX) induced testicular dysfunction. Mature rats received either MTX (20 mg/kg, intraperitoneally) or saline on the 7th day of experiment al design. Seven days prior and after MTX‐injection, rats received HA at dose of 300 mg/kg intragastrically (HA + MTX group; HA group alone). Serum was extracted and testicular tissues were examined for the changes in serum biochemistry (liver & kidney biomarkers, testicular hormones and antioxidants), molecular and histopthological alterations using RT‐PCR and immunohistochemistry. MTX‐injected rats induced alteration in all testicular parameters. Prior administration of HA ameliorated the MTX‐induced oxidative stress. HA administration normalised MTX‐induced decrease in serum levels of interleukin‐6 (IL‐6), tumour necrosis factor alpha (TNF‐α), interferon‐gamma (IFN‐γ), reproductive hormones (FSH, LH and testosterone) and antioxidants GST, SOD and catalase. MTX‐injected rats down‐regulated mRNA expression of GST, SOD, steroidogenesis associated genes, IFN‐γ, Bcl2 and NFKB. MTX up‐regulated BAX expression and caspase 9 immunoreactivity that were ameliorated in HA + MTX group. Collectively, HA ameliorated and restored all altered genes. In conclusion, HA is a promising supplement that is helpful in protection against testicular cytotoxicity and dysfunction induced by methotrexate.     

低温对大鼠睾丸扭转复位后生殖细胞凋亡的影响

目的 探讨低温对大鼠睾丸扭转复位后生殖细胞凋亡的影响。方法 24只健康青春期SD雄性大鼠随机分为三组:A组为睾丸扭转组,B组为睾丸扭转加低温组,C组为对照组。建立单侧睾丸扭转模型。术后第14天采集睾丸。原位缺口末端标记法(TUNEL)检测其生殖细胞凋亡指数(AI),光镜下观察睾丸组织学变化。结果 B组AI明显低于A组(P<0.01),而高于C组(P<0.01)。结论 低温能够提高扭转睾丸耐缺血能力,减少睾丸扭转复位后生殖细胞凋亡。     

茶多酚对大鼠睾丸扭转/复位模型保护作用的研究

目的:探讨茶多酚对大鼠睾丸扭转/复位模型的保护作用。方法:将24只健康雄性Wistar大鼠随机分为3组,每组8只。第Ⅰ组为假手术组(切开左侧阴囊游离睾丸,但不予扭转),第Ⅱ、Ⅲ组扭转左侧睾丸720°6h,分别于扭转复位前30min腹腔注射生理盐水和茶多酚,术后连续3d分别以低剂量维持。3组大鼠喂养至术后第5天处死,切取左侧扭转睾丸检测睾丸组织中超氧化物歧化酶(SOD)和丙二醛(MDA)含量;以原位缺口末端标记法(TUNEL)检测生精细胞凋亡指数(AI)。结果:Ⅰ、Ⅱ、Ⅲ3组左侧扭转睾丸组织SOD活力分别为(285.00±22.51)、(242.00±17.62)、(261.00±10.01)nU/mg;MDA含量分别为(1.81±0.20)、(4.34±0.34)、(2.94±0.38)nmol/mg;3组之间比较均有显著性差异。Ⅰ、Ⅱ、Ⅲ3组左侧扭转睾丸生精细胞凋亡指数(AI)分别为6.64±1.82、55.23±6.46、31.84±5.56,第Ⅲ组与第Ⅱ组相比,其生殖细胞凋亡明显减少(P<0.05)。结论:茶多酚对因睾丸扭转导致的缺血再灌注损伤具有保护作用。     

Protective effects of Ranolazine on testicular torsion and detorsion injury in rats

Ranolazine is a drug used in refractory chronic stable angina. In this study, it was aimed to evaluate the protective effect of ranolazine in a testis torsion model in light of objective biochemical and pathological data. A total of 24 pre-pubertal male Wistar albino rats were separated into three groups of 8 as the sham group, control group and ranolazine group. Testis torsion was applied for 3 hr to all the rats in Group Control and Group Ranolazine. In Group Control, 0.9% NaCl was applied 1 hr after the torsion. In Group Ranolazine, ranolazine 30 mg/kg was dissolved in a 0.9% NaCl solution and was administered intraperitoneally 1 hr after torsion. Histopathological evaluation was made using the Cosentino score. As a result of the objective biochemical and pathological criteria used in this study, this protective effect of ranolazine was observed in testis torsion. The results obtained in this study may suggest that ranolazine is a drug that could be applied after detorsion to patients diagnosed with torsion.     

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.     

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.