Mean Seminiferous Tubular Diameter Research Articles

The potential prophylactic and therapeutic impacts of niacin on ischemia/reperfusion injury of testis

The testicular ischemia-reperfusion (I/R) injury is characterized by the excessive aggregation of un-scavenged reactive oxygen species, leading to the heightened levels of oxidative stress. This phenomenon plays a pivotal role in the pathophysiology of testicular torsion damage. The current study aimed to detect the prophylactic and therapeutic effects of niacin on testicular I/R injury. Twenty-four healthy adult male Sprague Dawley rats were randomly allocated into three groups as follows: (1) sham group, (2) torsion/detorsion (T/D) group, and (3) treatment group which received 200mg/kg niacin along with testicular T/D. Torsion/detorsion was induced by 2h of torsion followed by 10 days of reperfusion period. In the treatment group, niacin was injected 30min before the reperfusion period intraperitoneally and continued for 10 days by oral gavage. T/D was associated with marked decreases in terms of sperm count, viability, and kinematic parameters versus the sham group (P<0.05), which niacin significantly reverted the kinematic parameters (P<0.05). I/R injury caused a significant increase in the number of abnormal epididymal sperms compared to the sham group (P<0.05). Niacin decreased the epididymal sperm abnormality significantly compared to the T/D group (P<0.05). Tissue abnormalities in T/D group, such as edema, hyperemia, inflammation, and necrosis were completely visible histopathologically, while the histological changes in the niacin-treated group were better than those in the T/D group. Regarding the pathological parametric evaluations, I/R injury significantly reduced the mean testicular biopsy score (MTBS), germinal epithelial cell thickness (GECT), and mean seminiferous tubular diameter (MSTD), and increased the tubular hypoplasia/atrophy (THA) compared to the sham group (P<0.05), which niacin treatment significantly improved the MTBS and GECT compared to the T/D group (P<0.05). T/D significantly increased the oxidative stress index (OSI) and lipid peroxidation (MDA) (P<0.05). Niacin significantly reduced the OSI and MDA levels compared to the T/D group (P<0.05). The current study found that niacin has preventive/therapeutic effects against the elevation of oxidative stress markers and depletion of antioxidants during I/R injury. Following administration of niacin, a reduction in histologic injury was observed in rats. In our study, we showed the antioxidant properties of niacin and its capacity to protect against I/R damage. The findings of the present investigation revealed that niacin, as an antioxidant agent, can suppress the oxidative stress induced by testicular I/R injury, and can be used as a supplementary agent in the treatment of those undergoing testicular torsion surgery.

The evaluation of citral effects on experimental unilateral testicular ischemia/reperfusion injury.

This investigation aimed to evaluate the defensive impacts of citral on ischemia/reperfusion (I/R) injury induced by testicular torsion/detorsion (T/D) in rats in an experimental model. The grouping of subjects was as follows: (1) sham, (2) T/D, (3) and (4) T/D plus citral 150 and 300 mg/kg, respectively, and (5) intact (citral 300 mg/kg). T/D was performed by testicular 720° turning for 2h and then detorsion for 24 h. Blood serum was obtained to assess testosterone and oxidative stress markers, epididymal sperms were collected for sperm staining and sperm analysis, and testicular tissues were examined for histopathology. T/D damage was associated with a remarkable decline in sperm total count, viability, and some velocity parameters in comparison to the sham group (p< 0.05), which could be reversed significantly by citral (p< 0.05). Histopathologically, T/D damage caused severe oedema, haemorrhage, inflammation, and seminiferous tubules disruption, while citral improved significantly the mean seminiferous tubular diameter, Cosentino's score, and Johnsen's score (p< 0.05). I/R injury was associated with significant increased malondialdehyde and oxidative stress index, and also significant reduced total antioxidant capacity and testosterone versus the sham group (p< 0.05), which all were prevented significantly by citral administration (p< 0.05). The outcomes greatly proved that testicular I/R injury could be significantly prevented by citral.

Age-related histomorphometric and ultrastructural changes in the Sertoli cells of Japanese quail (Coturnix Coturnix japonica)

Prepared sections from pre-pubertal, pubertal, adult, and aged Japanese quail testes were examined by light microscopy and transmission electron microscopy (TEM) and measurements of seminiferous tubular diameter (STD), luminal diameter (SLD), epithelial height (SEH) and cross-sectional area of the seminiferous tubules (AST) were taken using an image analyzer. Apoptotic Sertoli cells with features such as cell shrinkage and chromatin condensation were observed in pre-pubertal and aged quail. There was a significant difference between the mean Sertoli cell number (SCN), SLD, SEH, STD and AST among the four age groups (P < 0.001). The highest SCN (mean ± standard error) was recorded in the adult (30.53 ± 0.42), with the aged group displaying the lowest mean (11.80 ± 0.27) SCN. Spearman’s rho correlation coefficients demonstrated a strong relationship between the SCN and SEH in the pubertal (ρ=0.915; P < 0.001), adult (ρ=0.878; P < 0.001), and aged (ρ=0.858; P < 0.001) groups, while a significant moderate correlation was observed in the pre-pubertal (ρ=0.606; P < 0.001) group. There were significant moderate correlations between the SCN and STD in the pre-pubertal (ρ=0.445; P < 0.001), pubertal (ρ=0.653; P < 0.001), adult (ρ=0.440; P < 0.001), and aged (ρ=0.514; P < 0.001) groups. Furthermore, significant moderate correlations were estimated between the SCN and AST in the pre-pubertal (ρ=0.453; P < 0.001), pubertal (ρ=0.661; P < 0.001), adult (ρ=0.393; P = 0.001), and aged (ρ=0.498; P < 0.001) groups. This study provides baseline data on the morphology and development of the Sertoli cell, as well as testicular morphometry in avian species during the pre-pubertal, pubertal, adult, and aged stages using the Japanese quail as a model.

Protective effects of crocin on testicular torsion/detorsion in rats

Oxidative stress, caused by extreme accumulation of un-scavenged reactive oxygen species, plays an integral role in the Ischemia-Reperfusion (I/R) injury to the testicles following testicular torsion. The current research aimed to examine the protective effects of crocin as a natural antioxidant on testicular I/R injury in rats. Animals were divided randomly into five groups (seven each): (1) sham group, (2) torsion/detorsion (T/D) group, (3) intact group with 100 mg/kg crocin, (4) and (5) T/D groups followed by treatment with two different doses of crocin (50 and 100 mg/kg (IP)). I/R injury was induced by 720° clockwise torsion of the left testicles for 2 h. After 24 h of reperfusion, blood samples and epididymal sperms were collected to measure biochemical (GPx, SOD, and MDA), hormonal (testosterone), and sperm parameters (total sperm recovery, motility, viability, and morphology). Moreover, affected testicles were subjected to histopathology examination. I/R injury caused a significant reduction in sperm characteristics (except for morphology) (P < 0.05), which could not be significantly improved by crocin administration at either dose (P > 0.05). Johnsen's testicular score, mean seminiferous tubular diameter, and germinal epithelial cell thickness were significantly decreased in the T/D group compared to the intact and sham groups. However, crocin could significantly improve the histopathological parameters in both treatment groups compared to the T/D group (P < 0.05). T/D reduced SOD and GPx activity and testosterone level significantly (except for GPx) compared to the sham group (P < 0.05). However, crocin administration could significantly reverse them. Also, crocin reduced the amount of MDA significantly in the high-dose treatment group in comparison to T/D group (P < 0.05). The results of the current study revealed that crocin could be a promising agent to protect against I/R injury following surgical correction of the testicular torsion.

Leech therapy (Hirudo medicinalis) attenuates testicular damages induced by testicular ischemia/reperfusion in an animal model

BackgroundTesticular torsion/detorsion triggers tissue ischemia/reperfusion, leading to reactive oxygen species overgeneration and apoptosis. The saliva of leeches is full of anti-inflammatory, anticoagulants, antioxidants, and antimicrobial agents. Therefore, this study aimed to assess the protective mechanism of leech therapy on testicular ischemia/reperfusion damage.Methods18 adult male rats were randomly divided into three groups: 1-Sham-operated group (SO). 2-Torsion/detorsion (T.D) group: two hours of testicular torsion with two hours of testicular detorsion was performed. 3-Torsion/detorsion + Leech therapy (TDL) group. Sperm parameters (motility, vitality, morphology, and concentration), oxidative stress biomarkers (MDA, CAT, GPx, and TAC), histopathological factors (Mean seminiferous tubular diameter, Germinal epithelial cell thickness, Testicular capsule thickness, Johnson’s score, and Cosentino’s score), and immunohistochemical markers for apoptosis detection (Bax, Bcl-2, and Caspase-3) were measured.ResultsThere was a significant difference for all sperm parameters in the T. D group compared to the sham group. Leech therapy significantly increased progressive motility and normal morphology and reduced non-progressive motility. In the TDL group, MDA concentration significantly reduced, and levels of GPx, TAC, and CAT remarkably increased. All evaluated histopathological parameters in the TDL group significantly increased compared to the T. D group except for the testicular capsule thickness. T. D notably increased the expression of Bax and Caspase-3, while the treatment group slowed the rate of apoptosis compared to the control group. Bcl-2 expression in the T. D group was significantly lower than that in the sham group. Leech therapy increased the Bcl-2 expression.ConclusionLeech therapy attenuates damages to testicular tissue following torsion/detorsion due to its antioxidant, anti-inflammatory, and anti-apoptotic effects. Hence, it can be considered as an effective remedy for testicular ischemia/reperfusion.Graphical abstract

Testicular fixation and its effect on ipsilateral and contralateral testis in prepubertal rat model

BackgroundWhile performing orchidopexy, various suture materials or fibrin glues are used to achieve testicular fixation. This study was designed to assess the histological changes in testis after orchidopexy using fibrin glue and suture material.MethodsMale Wistar rats (n = 80) were divided randomly into four groups. Group I, (n = 20): sham operation, Group II (n = 20): Dartos Pouch (DP), Group III (n = 20): Transtunical fixation (TF), Group IV (n = 20): Tissue Adhesive (TA). Ipsilateral and contralateral testicular histology was assessed at 70 and 120 days of life after sacrificing animals by using thiopental sodium intraperitoneally at a dose of 100 mg/kg.ResultsMorphologically, at day 70, contralateral testis in Group III had a significant (p value 0.046) decrease testicular width (0.92 ± 0.01 vs 1.24 ± 0.39 cm). At 120 of life, Group I, II, III, and IV had a significant (p value < 0.001 each) decrease testicular width and weight in ipsilateral and decrease testicular length (p value 0.002) in contralateral testis. Histologically, mean seminiferous tubular diameter and DNA flow cytometry had a significant (p value < 0.001) decrease in size in Group I, II, III, and IV both ipsilateral as well as contralateral testis. Intergroup comparison at 70 and 120 days of life showed a significant decrease in seminiferous tubular diameter in Group II, III and IV and in Johnsen maturation score, seminiferous tubular diameter, DNA flow cytometry in Group I, II, III, and IV.ConclusionsDartos Pouch is most suitable procedure for treatment of orchidopexy. Suture fixation must be avoided and if the need arises then instead of suture materials, fibrin glue should be used for testicular fixation.

Administration of Dexmedetomidine Does Not Produce Long-Term Protective Effect on Testicular Damage Post Testicular Ischemia-Reperfusion Injury.

BackgroundAfter surgical correction of testicular torsion, up to 68% of ipsilateral testes undergo atrophy due to ischemia-reperfusion injury (IRI). Recent studies have shown that dexmedetomidine (Dex) alleviates IRI in various vital organs. However, those studies evaluated its protective effect on short-term reperfusion.PurposeWe aimed to investigate whether Dex has a long-term protective effect against testicular injury after IRI.Materials and MethodsA total of 24 New Zealand white rabbits were randomly divided into three groups (n = 8/group): the control group (saline-infused rabbits without IRI), the IRI group (saline-injected rabbits with IRI), and the Dex group (Dex-injected rabbits with IRI). The spermatic cord of rabbits in IRI and Dex groups was ligated for 4 h, and 1 h before reperfusion, Dex was administered intraperitoneally at a dose of 50 μg/kg body weight in group Dex, whereas saline was administered at the same dose to the IRI and control groups. Rabbits were kept alive for 4 weeks post reperfusion, then the testes were harvested, and the rabbits were euthanized.ResultsFour weeks post reperfusion, testicular volumes of the affected side decreased considerably in the IRI and Dex groups compared to the control group, with no significant difference between the IRI and Dex groups. Compared to the control group, the Johnson score and the mean seminiferous tubular diameters were significantly decreased in the IRI and Dex groups, but no significant differences were observed after administration of Dex. There were no significant differences in malondialdehyde and superoxide dismutase levels between the groups treated with and without Dex.ConclusionDex administration 3 h after ischemia and 1 h before reperfusion did not demonstrate a significant protective effect against testicular injury 4 weeks after IRI in rabbits. Further research is needed to confirm the potential therapeutic effects of Dex by varying the experimental conditions.

Histomorphometry of the Testes, Epididymides, and Spermatozoa of Four Indigenous Breeds of Bulls in Ibadan, Nigeria

Forty testicles were used to carry out histomorphometry study on the testes, epididymides and spermatozoa of four indigenous breeds of bull found in Ibadan (Red Bororo (RB), Sokoto Gudali (SG), White Fulani (WF) and Dhali (DL)). The testicles were harvested immediately after the slaughter at the Bodija abattoir and transported to the laboratory in an insulated flask containing warm water at 37?C within 30 minutes. In the laboratory, the samples were dissected and histological sections of the right testis, right and left caudal epididymides taken from two bulls in each group. The tissues were fixed in Bouin’s fluid. They were processed in an automatic tissue processor, embedded in paraffin wax using the embedding system (Leica EG 1160) and sectioned with microtome at 4 microns. The sections were then stained by Haematoxylin and Eosin method and mounted and examined under a microscope. There was no significant difference between breed and seminiferous tubular diameter (STD) and testicular germinal height (TGEH). The mean STD ranged from 223.00 ± 28.35 to 316.00 ± 37.70 μm, while the TGEH ranged from 81.60 ± 10.05 to 89.80 ± 4.83 μm. The mean epididymal tubular diameters (ETD) and epididymal lumen diameters (ELD) had the highest value of 378.00 ± 10.95 and 298.20 ± 26.0489 μm in DL and lowest of 373.80 ± 37.70 and 278.10 ± 27.71 μm in RB, respectively. The mean epididymal germinal epithelia heights (EGEH) were highest (61.20 ± 5.70 μm) in WF and lowest (39.80 ± 0.86 μm) in RB. The mean sperm head length (SHL), sperm midpiece length (SMPL), sperm tail length (STL) and sperm total length (TL) had the highest values of 5.90 ± 0.10, 8.80 ± 0.25, 24.20 ± 2.59 and 38.90 ± 2.94 μm, respectively in SG. SHL and SMPL were lowest in DL bulls (5.00 ± 0.00 and 6.30 ± 0.20 μm), while STL and TL were lowest (18.90 ± 0.24 and 31.30 ± 0.85 μm). The results of this study provide baseline data on the histomorphometry of the testes and epididymides and spermatozoa of some indigenous bulls.

Nitroglycerin ameliorates sperm parameters, oxidative stress and testicular injury following by testicular torsion/detorsion in male rats

PurposeThis study was designed to determine the probable protection mechanisms of nitroglycerin, a widely used medication for treatment of heart failure and angina, in amelioration of testicular ischemia/reperfusion damage. Methods24 adult male rats were randomly divided into three equal groups; with eight rats in each group: Group 1 (Sham) was sham-operated. Group 2 (T_D): 2 h testicular torsion was induced, afterward detorsion was performed and maintained for 2 h. Group 3 (NG): Nitroglycerin was administered immediately after detorsion. Sperm quality parameters such as viability, motility, morphology, and concentration, levels of antioxidant enzymes (glutathione peroxidase (GPX), catalase (CAT), and total antioxidant capacity (TAC)), and amount of malondialdehyde (MDA) in the blood plasma were examined in each group, thereafter histopathological parameters including germinal epithelial cell thickness (GECT), mean seminiferous tubular diameter (MSTD), Johnson's score and Cosentino's score were assessed. ResultsTesticular T_D significantly reduced sperm viability, motility, and normal morphology, whereas the NG administration remarkably increased the percentage of live, motile, and normal spermatozoa (p < 0.05). Levels of GPx, CAT, and TAC significantly reduced and the MDA level significantly increased in the T_D group in comparison to the sham group (p < 0.05). The NG treated group demonstrated significantly reduced MDA concentrations as well as elevated levels of GPx and CAT compared to the T_D group (p < 0.05). Induction of testicular torsion significantly reduced Johnson's score, GESCT (μm), and MSTD (μm), and remarkably increased the Cosentino's score (P < 0.05), while NG injection significantly increased Johnson's score, GESCT (μm), and MSTD (μm) and reduced the Cosentino's score (P < 0.05). ConclusionAccording to the findings in this research, nitroglycerin was able to protect the testicular tissue from ischemia-reperfusion damage caused by induced torsion/detorsion.

Investigating the sperm parameters, oxidative stress and histopathological effects of salvia miltiorrhiza hydroalcoholic extract in the prevention of testicular ischemia reperfusion damage in rats

AimsOne of the most common urologic emergencies is spermatic cord torsion, which can damage testicular tissue and reduce fertility. Salvia miltiorrhiza (SM) hydroalcoholic extract possess high antioxidant properties, and its efficacy in ischemia-reperfusion (I/R) injury prevention has been demonstrated in cardiac, renal, and liver tissues. Therefore, the purpose of this study was to assess the protective mechanism of SM extract on testicular I/R damage. Main methods18 mature male Wistar albino rats were randomly divided into 3 groups; with six rats in each group: Group 1 (Sham) was sham-operated. Group 2 (T-D): torsion was performed, and after 2 hours (h) detorsion was done. Group 3 (SM): (200 mg kg−1) SM was intraperitoneally injected thirty minutes before detorsion. Then testicular and epididymal weight and size alterations, sperm parameters (motility, livability, concentration, and morphology), both plasma and testicular tissue levels of malondialdehyde (MDA), catalase (CAT), glutathione peroxidase (GPX), and total antioxidant capacity (TAC) were evaluated. Also, histopathological changes included mean seminiferous tubular diameter (MSTD), testicular capsule thickness (TCT), mean testicular biopsy scoring (MTBS), and germinal epithelial cell thickness (GECT) were examined. ResultsTesticular I/R significantly reduced sperm motility, viability, and normality, while SM extract administration remarkably increased sperm motility, and normality (P < 0.05). Induction of testicular T-D caused a significant increment in the level of MDA and notable decline in the levels of GPX, CAT, and TAC both in plasma and testis tissue, whereas administration of SM extract significantly decreased MDA level and increased GPX, CAT, and TAC levels in plasma and testicular tissue (P < 0.05). Histopathological parameters including MSTD, GECT, MTBS, and TCT were significantly lower in the T-D group, while pretreatment with SM extract remarkably increased MSTD, GECT, and MTBS amounts (P < 0.05). ConclusionSince the SM extract increased the activity of antioxidant enzymes, improved sperm parameters and reduced the damage to testicular tissue, therefore, its use as a potent antioxidant in reducing testicular I/R damage is suggested.

Effects of L-carnitine and betamethasone on ischemia-reperfusion injuries and sperm parameters following testicular torsion in a rat model.

Testicular torsion is a consequence of spermatic cord twisting which causes progressive damage to the structure of the testis and reduces sperm quality and usually results in infertility. In the present study, with the assumption of the protective effects of L-carnitine and betamethasone against ischemia-reperfusion (IR) injuries, their effects on twisted testicles were evaluated and compared. Twenty Wistar rats were randomly divided into four groups and used in this study. Except for the Sham (S) group, testicular IR was induced surgically in three other groups, including Control (C), Betamethasone (BM), and L-carnitine (LC) groups. Betamethasone and L-carnitine were injected before detorsion in the BM and LC groups, respectively. After twelve hours of reperfusion, the testicles were detached, and prepared for sperm parameters evaluation such as sperm count, motility, viability, morphology, and chromatin quality, and histopathologic evaluations, including mean seminiferous tubular diameter (MSTD), germinal epithelial cell thickness (GECT), and Johnsen’s mean testicular biopsy scoring (MTBS). The MSTD, GECT, and healthy sperms in the C group were significantly lower than the other groups, while the BM and LC groups were significantly different from others in MTBS. The number of sperms and sperm motility in the BM group was significantly higher than the C group. Sperm viability in the BM and LC groups were significantly higher than the C group. The results of this study showed that both L-carnitine and betamethasone similarly can be effective in treating testicular IR injuries.

Omega-3 polyunsaturated fatty acids ameliorates testicular ischemia-reperfusion injury through the induction of Nrf2 and inhibition of NF-κB in rats

ObjectiveThis study was designed to investigate the protective effect of Omega-3 polyunsaturated fatty acids (n-3 PUFAs) on testicular ischemia-reperfusion (I/R) injury in rats. MethodsA total of 24 rats were randomly divided into the following three groups: Group A (Control group, n=8), Group B (I/R group, n=8), Group C (I/R group treated with n-3 PUFAs, n=8). Histological examination was used to assess changes in testicular structure. Tissue oxidative stress biomarkers, malondialdehyde (MDA) as well as Superoxide, and antioxidant indexes, including total antioxidant capacity (T-AOC); catalase (CAT); glutathione (GSH); glutathione/glutathione disulfide ratio (GSH/GSSG); superoxide dismutase (SOD) were determined. In addition, nuclear transcription factor erythroid 2-related factor 2 (Nrf2), Nrf2-dependent antioxidant enzymes, such as heme oxygenase-1 (HO-1) and NADPH quinine oxidoreductase-1 (NQO-1), and nuclear factor kappa B (NF-κB) were detected. ResultsCompared to I/R group, n-3 PUFAs could obviously increase the mean seminiferous tubular diameter in the histological examination. After n-3 PUFAs treatment, the level of tissue MDA and Superoxide were significantly decreased, while tissue T-AOC, CAT, GSH, GSH/GSSG and SOD levels were significantly increased, compared to I/R group (P<0.05). Besides, the expression levels of Nrf2, HO-1, and NQO-1 were significantly higher and the NF-κB expression level was significantly lower in the n-3 PUFAs treated group than that in I/R group (P<0.05). ConclusionsThese results provided evidences that n-3 PUFAs ameliorated testes damage caused by testicular I/R injury through its antioxidative capacity and anti-inflammatory effects, involving the activation of Nrf2 and the inhibition of NF-κB.

Hydrogen-rich saline attenuates spinal cord hemisection-induced testicular injury in rats.

To study how hydrogen-rich saline (HS) promotes the recovery of testicular biological function in a hemi-sectioned spinal cord injury (hSCI) rat model, a right hemisection was performed at the T11–T12 of the spinal cord in Wistar rats. Animals were divided into four groups: normal group; vehicle group: sham-operated rats administered saline; hSCI group: subjected to hSCI and administered saline; HRST group: subjected to hSCI and administered HS. Hind limb neurological function, testis index, testicular morphology, mean seminiferous tubular diameter (MSTD) and seminiferous epithelial thickness (MSET), the expression of heme oxygenase-1 (HO-1), mitofusin-2 (MFN-2), and high-mobility group box 1 (HMGB-1), cell ultrastructure, and apoptosis of spermatogenic cells were studied. The results indicated that hSCI significantly decreased the hind limb neurological function, testis index, MSTD, and MSET, and induced severe testicular morphological injury. The MFN-2 level was decreased, and HO-1 and HMGB-1 were overexpressed in testicular tissues. In addition, hSCI accelerated the apoptosis of spermatogenic cells and the ultrastructural damage of cells in the hypophysis and testis. After HS administration, all these parameters were considerably improved, and the characteristics of hSCI testes were similar to those of normal control testes. Taken together, HS administration can promote the recovery of testicular biological function by anti-oxidative, anti-inflammatory, and anti-apoptotic action. More importantly, HS can inhibit the hSCI-induced ultrastructural changes in gonadotrophs, ameliorate the abnormal regulation of the hypothalamic-pituitary-testis axis, and thereby promote the recovery of testicular injury. HS administration also inhibited the hSCI-induced ultrastructural changes in testicular spermatogenic cells, Sertoli cells and interstitial cells.

The histopathological evaluation of healing effects of vitamin C administered before methotrexate therapy on testicular injury induced by methotrexate.

Methotrexate (MTX) leads to acute toxic side effects in tissues or organs containing rapidly dividing cells such as seminiferous tubules. In this study, we investigated the protective effects of vitamin C against MTX-induced injury in rat testis. A total of 31 rats were divided into 4 groups, including the control group. The study was completed within 4 weeks and the rats received daily doses of 2 mL/kg SF, 100 mg/kg vitamin C and 10 mg/kg/day MTX i.p according to their groups. The mean seminiferous tubular diameter, germinal epithelial cell thickness, and mean testicular biopsy score were determined by histologic examination of each group. The vitamin C + MTX group showed more similarity with the control group. Statistically significant results were achieved between groups as for mean seminiferous tubular diameter, germinal epithelial cell thickness, and mean testicular biopsy score. When compared with the group which received vitamin C after MTX therapy, values for mean seminiferous tubular diameter, germinal epithelial cell thickness, and mean testicular biopsy score were significantly higher in the group which received vitamin C before initiation of MTX therapy. Vitamin C decreased MTX-induced testicular histological injuries, especially when used before MTX therapy.

The Effects of Local and Systemic Growth Hormone Treatment on Germ Cell Population and Fertility in an Experimental Unilateral Testicular Torsion and Orchiectomy Model

We evaluated the effects of local and systemic growth hormone on the germ cell population of the contralateral testes of pubertal rats subjected to unilateral testicular torsion and orchiectomy 24 hours later. A total of 40 male Wistar-Albino rats at age 3 weeks were divided into 5 groups. In the sham operated group the right testis was sutured and orchiectomy was performed 24 hours later. In groups 2 to 5 orchiectomy was performed 24 hours later following testicular torsion. In groups 3 and 4 unloaded and growth hormone loaded gelatin films, respectively, were sutured on the contralateral testes. In group 5 systemic growth hormone was administered for 7 days. Five weeks later each rat was cohabited with 2 female rats and the left testes were removed for evaluation. Mean seminiferous tubular diameter, mean testicular biopsy score and the mean haploid cell percentage were calculated. Mating studies were performed and fertility parameters were assayed. Mean seminiferous tubular diameter, mean testicular biopsy score and the mean haploid cell percentage of the contralateral testes were significantly decreased in the control and gelatin groups compared with the other groups. There was no difference between the local and systemic growth hormone groups regarding the haploid cell percentage. There were no differences between the groups in mean fetus numbers, mating or fertility and fecundity indexes except in the gelatin group, in which the mean fetus number was significantly lower. Fertility is not affected in rats after 24 hours of testicular torsion and orchiectomy, although there is germ cell injury and a decrease in the percent of haploid cells. Growth hormone administration resulted in the restoration of germ cell histology and an increase in the haploid cell percentage of the contralateral testes. Growth hormone may improve fertility after unilateral testicular torsion and orchiectomy.

Effects of heart of palm (Palmito) extract on reproductive system of adult male rats

To evaluate the toxicity effects of aqueous extract of heart of palm (Palmito) on plasma testosterone, estradiol, LH and FSH hormone levels and testicular tissue in male rats. A total of 70 male Sprague-Dawley rats were divided into seven groups. The control group was given 1 mL of distilled water, and the experimental groups were given 25, 50, 100, 150, 200, and 250 mg/kg/day of extract for 56 days. The effect of the treatment on the male reproductive system was evaluated by assessing plasma testosterone, estradiol, LH, and FSH levels, as well as spermatogenesis process. A significant decrease in testosterone and estradiol levels were observed in rats treated with different doses of palmito. The extract consumption provided no remarkable change in the serum levels of LH and FSH, and weights of bodies, and testes in the six treated-groups compared to those in the control group. Also, treatment with the palmito resulted in a decrease of the mean seminiferous tubular diameter and the number of spermatogonia, preleptoten and pachytene spermatocytes, round spermatids, and Leydig cells. The ethanol extract of palmito has anti-androgenic properties, which affected spermatogenetic activity in all experimental groups.

Comparison of streptozotocin-induced diabetic and insulin resistant effects on spermatogenesis with proliferating cell nuclear antigen (PCNA) immunostaining of adult rat testis

Male reproductive function is one of the mammalian systems that are impaired by diabetes mellitus and on the average, it will affect many more men prior to and during their reproductive years. The present study aimed to compare proliferating cell nuclear antigen (PCNA) immunostaining effects at stage VII of the spermatogenic cycle in streptozotocin-induced and insulin resistant diabetic rat testis. Male adult Sprague-Dawley rats (120-140 g) were randomly divided into 3 groups. Group 1: control group; fed on normal rat pellets. Group 2: streptozotocin diabetic group; received a single dose IP injection of streptozotocin 45 mg/kg BW in Na+ citrate buffer pH 4.5. Group 3: insulin resistant diabetic group; fed ad libitum on a special diet containing 25% fructose W/W. Following hyperglycaemia confirmation, animals were perfused with 4% Paraformaldehyde (PFA). Testes were isolated, and fixed in 4% PFA overnight, embedded in paraffin, 5μm thick sections were made and mounted on poly-L-lysine coated slides. Immunohistochemical staining was carried out on the positively charged slides using the PCNA as primary antibody.The results of this study showed that diabetes mellitus induced by streptozotocin injection and chronic fructose consumption through insulin resistance significantly (P<0.05) reduced PCNA index, mean seminiferous tubular diameter and testicular diameter. It could also be inferred from the results that diabetes mellitus and insulin resistance may necessarily not impair differentiation of primordial germ cells (gonocytes) into Type A and Type B spermatogonia. J. Exp. Clin. Med., 2012; 29:209-214

Comparison of the protective effect of dipyridamole and acetylsalicylic acid on long-term histologic damage in a rat model of testicular ischemia-reperfusion injury

PurposeIschemia reperfusion injury arising from testicular torsion results in a loss of spermatogenesis and a significant increase in germ cell apoptosis. We investigated the effects of dipyridamole and acetylsalicylic acid (ASA), 2 well-known platelet inhibitors, on testicular ischemia reperfusion injury. MethodsThirty adult male Sprague-Dawley rats were randomly divided into 5 groups (n = 6 for each group): control, sham-operated, torsion/detorsion (T/D), T/D + dipyridamole, and T/D + ASA. Testicular ischemia was achieved by rotating the left testis 720° clockwise for 2 hours. Thirty minutes before torsion, 10 mg/kg dipyridamole was injected transperitoneally in the T/D + dipyridamole group, and 100 mg/kg ASA was injected transperitoneally in the T/D + ASA group. Sixty days after the initial surgical procedure, ipsilateral orchiectomies were performed for histopathologic examination to determine Johnsen's mean testicular biopsy score (MTBS), mean seminiferous tubular diameter (MSTD), and apoptotic index (AI) in all groups. ResultsUnilateral testicular torsion-detorsion led to a significant decrease in Johnsen's MTBS and MSTD values in the ipsilateral testis and a significant increase in AI values of the T/D group. There were no significant differences between the T/D + dipyridamole and control groups in terms of MSTD and MTBS values. Although an amount of improvement exits in T/D + ASA group, there were significant differences between the T/D + ASA and control group MSTD and MTBS values. There was no significant difference between the T/D + dipyridamole and control groups in terms of AI values (P > .05), but the differences between the T/D + ASA and control groups were significant despite a slight decline in AI values of the T/D + ASA group. ConclusionsOur findings show that the use of dipyridamole before testicular reperfusion has a potentially protective effect against long-term injury in testicular ischemia reperfusion injury.

Protective Role of Hypothermia on Ischemia of Prepubertal Rodent Testicle

To assess the role of hypothermia in testicular ischemic injury in a prepubertal rat model. The study included 24 male, prepubertal Sprague-Dawley rats. Of the 24 rats, 20 were subjected to right testicular ischemia with and without hypothermia for 30 and 60 minutes, 5 in each group. The remaining 4 rats underwent sham operation and were used as controls. A vascular clamp was used to clamp the spermatic cord for the specified time. Hypothermic treatment consisted of placing the testicle in ice slush. At 8 weeks postoperatively, both testicles were harvested. The left testicle was used as the control. A pathologist, who was unaware of the groups, examined all the slides. The mean seminiferous tubular diameter (STD) and mean number of germinal cell layers (GCLs) were obtained. When the contralateral testicle was used as the control, the mean GCL and STD in the ipsilateral operated sides were significantly worse in all ischemic groups. When the sham-operated testicles were used as the control, no significant differences were found between the cold ischemia groups and the sham group. In the warm ischemia groups, the mean GCL and STD were significantly worse than those in the sham group. We compared the operated sides among the groups. At 30 minutes, the mean GCL and STD showed a trend toward preservation with cold ischemia, although the difference was insignificant. At 60 minutes, the mean GCL and STD were significantly worse with warm than with cold ischemia. The results of our study have shown that permanent ipsilateral ischemic testicular damage occurred as early as 30 minutes in prepubertal rats. The damage might be reduced with hypothermia.

Role of serum- and glucocorticoid-inducible kinase-1 in regulating torsion-induced apoptosis in rats

Serum- and glucocorticoid-inducible kinase-1 (SGK1) is a serine/threonine protein kinase that responds to various stimuli and mediates cell survival. Although it is known that testicular torsion leads to testicular damage and male infertility, the role of SGK1 in torsion remains unclear. This study investigated whether torsion-induced apoptosis is associated with changes in phosphoinositide-dependent protein kinase-1 (PDK1), SGK1 and forkhead transcription factor FOXO3a expression and/or phosphorylation in rats. Sprague-Dawley rats were divided into four groups: sham (control), 1, 2 and 4 h of unilateral torsion. Bilateral testes, testicular interstitial fluid (TIF) and blood samples were collected immediately after torsion. Our results revealed that SGK1 protein and mRNA were abundantly present in testes and were induced by 2 h of torsion, but that phosphorylation of SGK1, PDK1 and FOXO3a decreased simultaneously. After 2 h of torsion, the testosterone secretion capacity of the primary Leydig cells and testicular interstitial cells (TICs) was impaired and apoptotic spermatogonia and TICs were observed; in addition, the mean seminiferous tubular diameter was decreased. Torsion increased plasma corticosterone levels, but decreased plasma luteinizing hormone and testosterone levels. However, the testosterone levels of the TIF in the ipsilateral testes were significantly enhanced after 2 h of torsion, but suppressed in the contralateral testes. This animal study suggests that PDK1, SGK1 and FOXO3a are involved in torsion-induced apoptosis and that medical therapy should be performed as early as 2 h after the occurrence of torsion to prevent further damage.