Experimental Varicocele Model Research Articles

Shugan Tongluo Qiangjing recipe protects against varicocele of EVC rats through modulating sperm DNA damage, telomere expression and oxidative stress

Varicocele (VC) refers to expansion and tortuosity of spreading venous plexus in spermatic cord due to poor blood flow. This study aimed to investigate effects of Shugan Tongluo Qiangjing recipe (SGTL) on sperm DNA damage and oxidative stress in experimental VC (EVC) rats. EVC model was established by partial ligation of left renal vein. Spermatic vein diameter, testicular weight, sperm DNA fragmentation index (DFI) were evaluated. Telomere reverse transcriptase (TERT) expression, telomere gene transcription, and testicular tissue morphology were determined·H2O2, catalase, SOD, T-AOC were measured with colorimetry. SGTL significantly decreased spermatic vein diameter (P=0.000) and increased testicular weight (P=0.013) of rats compared those of EVC rats. SGTL maintained testicular tissue morphology in EVC rats. SGTL markedly reduced sperm DFI value in sperm of rats compared to EVC rats (P=0.000). SGTL significantly enhanced TERT expression and telomere gene transcription (P=0.028) in testis of rats compared to EVC rats. SGTL reduced H2O2 levels (P=0.001) and promoted CAT activity (P=0.016), SOD activity (P=0.049), and T-AOC activity (P=0.047) of rats, compared to EVC rats. In conclusion, SGTL could reduce pathogenic process of EVC by reducing sperm DNA damage and regulating telomere length in EVC rats, which may be related to oxidative stress regulation.

Effect of vitamin D in experimental varicocele model in rats

Purpose: Varicocele is a pathological condition that affects approximately 20% of adult men, causing infertility and sperm deterioration. The aim of our study was to investigate the efficacy of vitamin D (Vit. D) in the pathophysiology of varicocele. We performed biochemical, gene expression analyses and histopathological to evaluate the efficacy of vitamin D in the experimental varicocele model.
 Materials and Methods: In the study, 30 adult male Wistar Albino rats were used. The rats were divided into 3 groups equally as control group, experimental group (varicocele), treatment group (varicocele + D vit.). The treatment group received 500 IU/kg D vit. intramuscularly.
 Results: Histopathological, TRPM2-8 gene expression and biochemical analyses were performed on testicular and blood samples collected at the end of the experiment. The experimental group showed a deterioration in tubular structure, a decrease in total antioxidant levels and an increase in total oxidant levels. The treatment group, on the other hand, showed an increase in TAS, a decrease in TOS and a beneficial improvement in tubular structure disorders. Analysis of gene expression levels showed that TRPM2-8 expression levels were significantly increased in the varicocele group and decreased in the treatment group.
 Conclusion: In the varicocele model, the use of vitamin D had a significant effect on TRPM2-8 gene level, pathological seminiferous tubules and biochemical values. Further studies are needed to determine the clinical application of vitamin D in varicocele disease.

Zishen Yutai Pill improves sperm quality and reduces testicular inflammation in experimental varicocele rats

ObjectiveZishen Yutai Pill (ZYP), containing 15 Chinese traditional medicine, is a safe and well quality-controlled TCM preparation with promising effects in many fields of reproduction. The current study was designed to investigate the therapeutic effects of ZYP on sperm quality and testis in varicocele (VC) rats. Materials and methodsMale Wistar rats were randomly divided into four groups (n = 6), i.e., a sham group, a VC group, and VC groups treated with different dose of ZYP (1575 and 3150 mg/kg/d, respectively). The experimental VC model was established by partial ligation of left renal vein. Six weeks after model establishment, ZYP was orally administered once a day for the next 6 weeks. Parameters relating to testis and sperm quality were assessed. Hematoxylin–eosin staining was used to showed testicular tissue damage in experimental VC rats. Expressions of proteins relating to NLRP3 inflammasome pathways were determined using Western blot (WB). The mRNA expressions of relating genes were determined using quantitative real-time PCR (qRT-PCR) analysis. ResultsZYP could significantly improve sperm motility and decrease sperm DNA fragmentation index in VC rats (P < 0.05). Hematoxylin-eosin (HE) staining showed that ZYP could alleviate testicular tissue damage caused by experimental varicocele in rats. Compared to the VC model, expressions of NLRP3, ASC, and caspase-1 in rats treated with ZYP were significantly down-regulated, as validated by both qRT-PCR and WB analysis (P < 0.05). ConclusionsIn brief, ZYP could improve sperm DNA integrity by inhibiting the NLRP3 inflammasome pathway and alleviating the chronic inflammation of testicular tissue induced by experimental varicocele in rats.

Beneficial effects of combination therapy with testosterone and hydrogen sulfide by reducing oxidative stress and apoptosis: Rat experimental varicocele model.

Despite the effectiveness of testosterone therapy in conditions associated with testosterone deficiency, including varicocele, several dose-dependent side effects limit the clinical use of testosterone therapy. Hydrogen sulfide, a toxic gas in high concentrations but a beneficial molecule in low concentrations, acts as both a major effector and an important inducer of testosterone. This study investigated whether a subeffective dose of testosterone combined with a subeffective dose of hydrogen sulfide donor sodium hydrosulfide (NaHS) can be effective in an experimental varicocele model through a possible additive effect. Thirty Wistar rats weighing 200-250 gr were divided into 5 groups as (n = 6/each): sham, varicocele, testosterone (200 µg/kg, 5 times per wk for 4 consecutive weeks), NaHS (15 μmol/L, daily for 4 consecutive wk) and testosterone + NaHS (200 µg/kg, 5 times per wk + 15 μmol/L, daily, both for 4 consecutive wk). All animals, except in the sham group, underwent varicocele induction. The coadministration of testosterone and NaHS significantly increased serum testosterone (10.23 0.95, p = 0.01), testicular H2S levels (608.94 21.09, p 0.001), and testicular superoxide dismutase activity (66.14 1.56, p 0.001), decreased malondialdehyde levels (0.77 0.52, p 0.001), and B-cell lymphoma 2-associated X protein to B-cell lymphoma 2 (0.16 0.01, p 0.001) protein expression ratio in the testicular tissues and improved sperm parameters and testicular histopathology compared to the varicocele group. The combination therapy of subeffective doses of testosterone and NaHS can attenuate the varicocele-induced damages by reducing testicular oxidative stress and apoptosis and thus can be considered an effective approach with fewer side effects.

Silencing CAMK2D Promotes the Proliferation of Spermatogonia in the Testis of Experimental Varicocele Rats.

Varicocele is regarded as the main factor that contributes to male infertility. This study aimed to explore the effect of CAMK2D on spermatogonia in the testis of experimental varicocele rats. The experimental varicocele model was established in rats and treated using different ligation methods. mRNA expression profile analysis was performed on the left testicular tissue isolated from different groups, and differentially expressed genes (DEGs) were analysed by bioinformatics methods and identified by qRT-PCR. The effect of CAMK2D, the screened DEG, on the proliferation of spermatogonia was evaluated by CCK-8 assay. The expression level of the c-kit was measured by the immunofluorescence assay and the expression levels of CAMKII, FOXO1, and β-catenin were detected by qRT-PCR and western blotting. Five DEGs (i.e., TMCC3, FLNB, CAMK2D, OPLAH, and EGR1) were screened using the comprehensive analysis of mRNA high-throughput sequencing data. TMCC3 and FLNB were significantly downregulated, and CAMK2D, OPLAH, and EGR1 were dramatically upregulated in the testicular tissue of varicocele rats. The target DEG CAMK2D was obtained through identification by using qRT-PCR. In vitro assays revealed that the proliferation of spermatogonia was significantly facilitated by the silencing of CAMK2D, which resulted in the downregulation of CAMKII, FOXO1, and β-catenin. In conclusion, silencing CAMK2D facilitated the proliferation of spermatogonia in the testis of experimental varicocele rats.

Lycopene protects sperm from oxidative stress in the experimental varicocele model.

Oxidative stress (OS) is an important parameter in the evaluation of infertility caused by varicocele. Antioxidants are the most commonly prescribed drugs in these patients. Lycopene molecule, as the powerful antioxidant in the carotenoid family, has beneficial effects on improving fertility in males. Therefore, we investigated the effects of lycopene on induced OS by varicocele in an animal model. Forty‐five adult male Wistar rats were divided into two groups: control (n = 12) and varicocele (n = 33). Two months after induced varicocele, five rats in each group were sacrificed randomly and induced varicocele was investigated. Remained rats were divided into five groups (n = 7), including the control (I), varicocele (II), varicocele reserving solvent (III), varicocele reserving lycopene 4 mg/kg (IV), and 10 mg/kg (V) for two months. At the end of the experiment, intracellular reactive oxygen species (ROS), malondialdehyde (MDA), total antioxidant capacity (TAC), %DNA damage, and antioxidant enzymatic levels were measured. The results indicated that there were significant increases in the levels of ROS, MDA, DNA damage, superoxide dismutase (SOD), sperm concentration, and motility in the varicocele groups compared with the control group. In the lycopene group (10 mg/kg), sperm concentration, the levels of TAC, and catalase (CAT) activity were improved so the levels of ROS, MDA, and %DNA damage were reduced compared with varicocele group. Our findings indicated that the administration of lycopene especially at a dose of 10 mg/kg in the varicocele group could protect sperm from OS and sperm DNA damage by increasing antioxidant activity and reducing ROS.

NLRP3 Inflammasome: A New Pharmacological Target for Reducing Testicular Damage Associated with Varicocele.

Many bioactive natural compounds are being increasingly used for therapeutics and nutraceutical applications to counteract male infertility, particularly varicocele. The roles of selenium and Polydeoxyribonucleotide (PDRN) were investigated in an experimental model of varicocele, with particular regard to the role of NLRP3 inflammasome. Male rats underwent sham operation and were daily administered with vehicle, seleno-L-methionine (Se), PDRN, and with the association Se-PDRN. Another group of rats were operated for varicocele. After twenty-eight days, sham and varicocele rats were sacrificed and both testes were weighted and analyzed. All the other rats were challenged for one month with the same compounds. In varicocele animals, lower testosterone levels, testes weight, NLRP3 inflammasome, IL-1β and caspase-1 increased gene expression were demonstrated. TUNEL assay showed an increased number of apoptotic cells. Structural and ultrastructural damage to testes was also shown. PDRN alone significantly improved all considered parameters more than Se. The Se-PDRN association significantly improved all morphological parameters, significantly increased testosterone levels, and reduced NLRP3 inflammasome, caspase-1 and IL-1β expression and TUNEL-positive cell numbers. Our results suggest that NLRP3 inflammasome can be considered an interesting target in varicocele and that Se-PDRN may be a new medical approach in support to surgery.

Nutraceutical Effects of Lycopene in Experimental Varicocele: An "In Vivo" Model to Study Male Infertility.

Varicocele is one of the main causes of infertility in men. Oxidative stress and consequently apoptosis activation contribute to varicocele pathogenesis, worsening its prognosis. Natural products, such as lycopene, showed antioxidant and anti-inflammatory effects in several experimental models, also in testes. In this study we investigated lycopene effects in an experimental model of varicocele. Male rats (n = 14) underwent sham operations and were administered with vehicle (n = 7) or with lycopene (n = 7; 1 mg/kg i.p., daily). Another group of animals (n = 14) underwent surgical varicocele. After 28 days, the sham and 7 varicocele animals were euthanized, and both operated and contralateral testes were weighted and processed. The remaining rats were treated with lycopene (1 mg/kg i.p., daily) for 30 days. Varicocele rats showed reduced testosterone levels, testes weight, Bcl-2 mRNA expression, changes in testes structure and increased malondialdehyde levels and BAX gene expression. TUNEL (Terminal Deoxynucleotidyl Transferase dUTP Nick End Labeling) assay showed an increased number of apoptotic cells. Treatment with lycopene significantly increased testosterone levels, testes weight, and Bcl-2 mRNA expression, improved tubular structure and decreased malondialdehyde levels, BAX mRNA expression and TUNEL-positive cells. The present results show that lycopene exerts beneficial effects in testes, and suggest that supplementation with the tomato-derived carotenoid might be considered a novel nutraceutical strategy for the treatment of varicocele and male infertility.

Effect of royal jelly on testicular antioxidant enzymes activity, MDA level and spermatogenesis in rat experimental Varicocele model

Varicocele is one of the most prevalent causes of infertility. It causes induction of oxidative stress, increases lipid peroxidation in the testis and disrupts spermatogenesis cycle. The aim of the study was to investigate the possible protective effects of royal jelly against varicocele induced oxidative stress, biochemical and histological alterations in the experimental varicocele model in rat. Twenty-one adult Wistar rats were divided into three groups. The control group (I), Varicocele and administration of normal saline (II), varicocele and treatment with RJ (III). At the end of the experiment, all the animals were sacrificed and testes excised. The activity of catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx) and MDA levels were measured. Also, histopathological examinations, Johnsen scores and sperm parameters were determined. There was a significant (p<0.05) increase in the activity level of CAT (0.223±0.005), SOD (0.177±0.0062), GPx (9.575±0.318) and a significant (p<0.05) reduction in the MDA level (2.674±0.336) of the experimental varicocele treated with royal jelly when compared to the activity of CAT (0.011±0.004), SOD (0.035±0.0096), GPx (8.864±0.397) and MDA level (4.630±0.579) of the experimental varicocele and administration of normal saline. Results of the Johnsen score showed a significant increase (p<0.05) in the mean score of the RJ group (7.94±1.5) when compared to the normal saline group (6.04±1.4). Therefore, RJ is a potential area for further studies and improving in spermatogenesis cycle after varicocele.

Protective effects of HSP70, HO-1 and HSP27 on spermatogenic function in rats with varicocele

Objective To investigate the protective effects of HSP70、HO-1 and HSP27 on spermatogenic function in rats with varicocele. Methods Forty adult male SD rats were randomly divided into 4 groups with 10 rats in each group: sham group (group A), sham + quercetin group (group B), VC group (group C) and VC+ quercetin group (group D). An experimental varicocele model was successfully created in group C and D and quercetin(100 mg/kg) was injected intraperitoneally. HE staining was used to observe the pathological changes of testis. Malondialdehyde (MDA) and superoxide dismutase (SOD) content were determined. Cell apoptosis index was detected by dUTP nick end labeling. The expression of HSP70 in testis tissue was detected by immunohistochemistry. The mRNA expression of HSP27 and HO-1 in testis tissue was measured by RT-PCR. Results Testicular structure and spermatogenic function were seriously affected, and histological damage was significantly increased caused by VC in group D. AI was (5.26±0.45)% , (5.83±0.52)% , (14.64±0.98)% , (25.37±1.46)%, respectively.The SOD activities were(0.96±0.06), (0.88±0.05), (0.45±0.04), (0.23±0.03) U/mg protein, MDA content was(2.24±0.35), (2.69±0.32), (4.78±0.41), (7.37±0.68) nmol/g protein.The expression of HSP70 in group C was significantly higher than that in group A and B, and the expression of HSP70 in group D was significantly lower than that of group C. Compared with group A and B, the mRNA expression level of HSP27 and HO-1 was significantly increased in group C. Compared with group C, the expression level of HSP27 and HO-1 in group D was significantly decreased.The comparison between B、C and D groups was statistically significant (P<0.05). Conclusions HSP70、HO-1 and HSP27 reduces the level of oxidative stress and inhibits the apoptosis.Thus, it plays a protective role on the spermatogenic function in rats with varicocele. Key words: Varicocele; Disease Models, Animal; HSC70 Heat-Shock Proteins; Heat-Shock Proteins; Rats

Comparative Expression Analysis of HSP70, HSP90, IL-4, TNF, KITLG and KIT-receptor Gene between Varicocele-Induced and Non-Varicocele Testes of Dog.

This study was designed to create an experimental varicocele model by a simple surgical procedure in dog with minimum invasion and to investigate the effect of varicocele-induced infertility on the expression of six related genes (HSP90, HSP70, IL- 4, TNF, KITLG and KIT receptor). In this experimental study, the proximal part of the pampiniform plexus of dog testes was partially occluded without abdominal incision which was confirmed by venographic examination. To evaluate varicocele in its acute form, dogs were castrated after 15 days and testes were dissected. Histopathologic evaluation was undertaken and the relative expression of the six genes was assessed by quantitative realtime polymerase chain reaction (PCR). Microscopic changes showed tubule degeneration. The Johnson score was significantly decreased in the varicocele testes when compared with non-varicocele testes. Expressions of HSP90, TNF, KITLG and the KIT-receptor gene were significantly downregulated (P=0.029, 0.047, 0.004 and 0.035 respectively) in varicocele-induced testes while HSP70 was upregulated (P=0.018). IL-4 did not show differential expression (P=0.377). We conclude that partial occlusion of the proximal part of the pampiniform plexus induces varicocele in the testis of dog. Differential expression of the mentioned genes may be responsible for the pathophysiology of varicocele and related subfertility.

Role of Inhibitors of Apoptosis Proteins in Testicular Function and Male Fertility: Effects of Polydeoxyribonucleotide Administration in Experimental Varicocele.

Neuronal apoptosis inhibitory protein (NAIP) and survivin might play an important role in testicular function. We investigated the effect of PDRN, an agonist of adenosine A2A receptor, on testicular NAIP and survivin expression in an experimental model of varicocele. After the creation of experimental varicocele (28 days), adolescent male Sprague-Dawley rats were randomized to one of the following treatments lasting 21 days: vehicle, PDRN (8 mg/kg i.p., daily), PDRN + 3,7-dimethyl-propargylxanthine (DMPX, a specific adenosine A2A-receptor antagonist, 0.1 mg/kg i.p., daily), varicocelectomy, and varicocelectomy + PDRN (8 mg/kg i.p., daily). Sham-operated animals were used as controls. Animals were then euthanized and testis expression of NAIP and survivin was evaluated through qRT-PCR, western blot, and immunohistochemical analysis. Spermatogenetic activity was also assessed. NAIP and survivin expressions were significantly reduced following varicocele induction when compared to sham animals whereas PDRN-treated rats showed an increase in NAIP and survivin levels. Immunohistochemistry revealed an enhanced expression of NAIP and survivin with a characteristic pattern of cellular localization following PDRN treatment. Moreover, administration of PDRN significantly restored spermatogenic function in varicocele rats. PDRN may represent a rational therapeutic option for accelerating recovery from depressed testicular function through a strategic modulation of apoptosis in experimental varicocele.

Different surgical techniques and L-carnitine supplementation in an experimental varicocele model

We aimed to investigate the impact of various varicocelectomy techniques and/or L-carnitine as an adjunct treatment, following the emergence of oxidative stress, on the expression levels of SCF/c-kit signalling pathways in spermatogenesis. Forty-two rats were divided into seven groups: group 1 (G1) control; group 2 (G2) sham; group 3 (G3) varicocele; group 4 (G4) varicocele + varicocelectomy with testicular nonartery sparing; group 5 (G5) same as G4 but with artery sparing; group 6 (G6) same as G4 but with L-carnitine and group 7 (G7) same as G5 with L-carnitine. mRNA expression levels of SCF and c-kit were measured quantitatively using real-time polymerase chain reaction. CASP-3 activity at protein level was determined, and histological evaluation was performed. mRNA expression level of SCF increased in G6 as compared to control group (3.52-folds change; P = 0.035), whereas mRNA expression level of c-kit gene remained the same. We found that in the left testis of G6 group, mRNA expression level of SCF increased 2.2-folds in comparison with the right testis (P < 0.05). There were no statistically significant differences in the CASP-3 protein expression levels between the control and other groups. When Cosentino Score analyses of immunostaining were conducted, we observed no significant differences among groups. Spermatogenic failure could be primarily due to a sertoli cell dysfunction. Although surgical treatment has been the best option for management of varicocele, auxiliary agents like L-carnitine may be considered as supportive treatment regimes in addition to conventional surgical treatments.

S1 THE IMPACT OF UNILATERAL EXPERIMENTAL RAT VARICOCELE MODEL ON TESTICULAR HISTOPATHOLOGY, LEYDIG CELL COUNTS, AND INTRATESTICULAR TESTOSTERONE LEVELS OF BOTH TESTES

Varicocele, most treatable pathologic condition in male infertility, exerts unfavorable effects on testicular ultrastructure via various mechanisms. In this study we aimed to demonstrate adverse effects of varicocele on both testes.Twenty one adult male Albino rats were divided into 3 groups. Sham operation was performed for group 1 (control group), and this group of rats were sacrificed 4 weeks later. Experimental varicocele model was performed for group 2 (varicocele group) and these animals were sacrificed 4 weeks after the operation. In group 3 the rats were varicocelectomized 4 weeks later. This group of rats were sacrificed at 4 weeks postoperatively. The level of testicular damage was examined, and serum testosterone and intratesticular testosterone levels were measured.Mean (±SD) damage scores of the right testes of the sham, varicocele, and varicocelectomy groups were 0, 1.64 ± 1.3, and 1.21 ± 0.3, respectively. There was no statistically significant differences between damage scores of groups 2, and 3 (P = .320), relevant scores of both groups were determined to be significantly higher than group 1 (P = .009, and P = .001). Mean (±) damage scores of the left testes of the three groups were detected to be 0.43 ± 1.13, 2.29 ± 1.15, and 1.78 ± 0.39, respectively. The difference between varicocele, and varicocelectomy groups was not statistically significant (P = .112).Unilateral varicocele has deleterious effects on both testes. There was no statistically significant difference as for histopathologic recovery following varicocelectomy.

Effects of Angiogenesis Inhibition by Spironolactone on Isolated Vas Deferens Contractility in an Experimental Varicocele Model in Rats

To investigate the effect of spironolactone, as an angiogenesis inhibitor, on the isometric contractile responses in isolated vas deferens strips from left varicocele-induced rats. Twenty-four adult (12-14 months) male Wistar albino rats were randomly assigned to 4 groups (n = 6 in each): (1) Control group, (2) sham-operated group, (3) experimental left varicocele group, and (4) Spironolactone (20 mg/kg/d)-treated experimental left varicocele group. Histopathologic and immunohistochemical (CD31 staining) findings in the rat testis and functional findings in the rat isolated vas deferens were investigated. Angiogenesis increased in the varicocele group and the spironolactone inhibited angiogenesis in the spironolactone-treated group. Spironolactone seemed to change phenylephrine and serotonin responses in the left vas deferens. It is possible that by inhibiting angiogenesis, spironolactone treatment negatively impairs testicular morphology and functional (vas deferens) pathways. Varicocele formation seems to elicit an increase to 5-HT sensitivity in rat vas deferens, and this process is prevented by spironolactone pretreatment.

Potential role of p38-mitogene-activated protein kinase and nuclear factor-kappa B expression in testicular dysfunction associated with varicocele: an experimental study

The aim of this study was to investigate p38-mitogene-activated protein kinase (p38-MAPK), nuclear factor-kappa B (p65-NF-kB) and inducible nitric oxide synthase (iNOS) expression in an experimental model of varicocele in the rat testis. Male Wistar albino rats (n = 18) were divided into three equal groups: control group, sham operated group and left varicocele-induced group. Malondialdehyde (MDA), nitric oxide (NO) and reduced glutathione (GSH) levels were biochemically assessed, and the p38-MAPK and NF-kB activity, and iNOS expression were immunohistochemically studied in the right and left testicles of rats from each group. The GSH levels were significantly decreased, whereas the level of MDA and NO was significantly increased in the testicular tissues of rats in varicocele group compared with those of the control and sham groups. There was a marked staining for iNOS, p38-MAPK and p65-NF-kB expression in rats of varicocele group compared with the sham group. There was no positive staining in rats of control group. There were significant differences in biochemical, histological and immunohistochemical studies, but no significant differences were noted between other groups. p38-MAPK and p65-NF-kB activation, and iNOS expression have a significant role in varicocele-induced testicular dysfunction.

The effects of N-acetylcysteine on spermatogenesis and degree of testicular germ cell apoptosis in an experimental model of varicocele in rats

The mechanism by which varicocele interferes in spermatogenesis has not been clearly defined. Germ cell apoptosis and oxidative stress appear to be involved in this process and the use of antioxidants has been proposed to counteract upon these effects. The present study evaluated the effects of N-acetylcysteine (NAC) on spermatogenesis and germ cell apoptosis in an experimental model of varicocele in rats. Twenty 30-day-old animals were randomly divided into three groups: sham operation (Group 1), left experimental varicocele (Group 2) and left experimental varicocele group treated with NAC 50 mg/kg/day (Group 3). After 2 months, spermatogenesis was evaluated by absolute and true count of round spermatids, pachytenes, spermatocytes and Sertoli cells. The different cell relations were also analyzed. Germ cell apoptosis was quantified using the TUNEL method. The apoptotic index (AI) was calculated as the number of apoptotic cells per tubule. Statistical analysis was performed by analysis of variance considering P < 0.05. The absolute and true cell counts were similar among the groups (P > 0.05). The round spermatid/pachytene ratio was significantly smaller in Groups 2 and 3 compared to the Group 1 (P = 0.012). The AI values were 0.207 ± 0.09, 0.138 ± 0.11 and 0.298 ± 0.27, respectively (P = 0.256). Experimental varicocele in rats presented an association with the decreased round spermatid/pachytene ratio, suggesting the loss of germ cells during spermatogenesis. These effects were not influenced by the administration of NAC. Germ cell apoptosis was not influenced by experimental varicocele.

A new experimental inbred Wistar rat varicocele model: anatomy of the left spermatic vein and the effect on histology

Because of venous anatomical differences between rats and humans and the personal interpretation of these differences, there is neither consistent animal prototype nor consistent results in the study of varicocele. We established a new substrain of Wistar inbred rats, of which the left testis vein has no significant branches to the common iliac vein up pampiniform plexus, but instead enters the left renal vein directly (similar to humans) and used them to create experimental varicocele model by partial ligation of the left renal vein. One month later, the predominant lesion of the left testis in induced group was spermatogenic arrest at the spermatid and preliminary spermatocyte phases, and considerable interstitial and Sertoli cell vacuolation. The right testis also showed spermatogenic arrest. Most important, the characteristics of the lesions differed in both testes, with the left testis having more severe lesions. Allowing for the unique anatomy of the left spermatic vein, the standard of the surgical procedure, the high rate of varicocele induction, and identical histological alteration as occurs in humans, we believe that this inbred Wistar rat substrain is suitable for the creation of an experimental varicocele model, which has promise for practical application in humans.

The effects of varicocele and its surgical correction on vas deferens motility

Varicocele is the abnormal dilation of venous pampiniform plexus and internal spermatic vein. Its prevalence in the adolescent period is almost equal to the prevalence of adult age. That is why the disease is accepted to appear in early adolescence and does not disappear spontaneously. Varicocele is established to be the most common cause of infertility in the adulthood period in terms of the testicular and/or epididymal damages it causes. Besides, malfunctioning of testis and/or epididymis cannot be blamed as the one and only reason of infertility. One major reason of the male infertility is vas deferens motility disorders. There is limited data in the literature investigating the effects of varicocele on the vas deferens motility. The aim of the study is to evaluate not only the motility defects of vas deferens for the period of varicocele, but also the effects of surgical varicocele correction on vas deferens motility. Thirty male Wistar-Albino rats were allocated to five groups. In the control group (Gr C, n = 6) bilateral vas deferens strips were harvested without any surgical intervention. Using the partial left renal vein obstruction technique, the experimental varicocele model was performed for the other four groups. Varicocele was apparent for these animals after the fourth week of the venous ligation. Bilateral vas deferens strips of varicocele group (Gr V, n = 6) were harvested. The rest of the animals having varicocele underwent relaparotomies. Three different surgical procedures were performed to these animals. The animals of group P (Gr P, n = 6) and group I (Gr I, n = 6) underwent Palomo and Ivanissevich procedures, respectively, for varicocele correction. And the animals of group S (Gr S, n = 6) underwent sham operation. After 4 weeks of relaparotomies, bilateral vas deferens strips of all three groups harvested. The electrical field stimulation (EFS) induced responses of all vas deferens strips as well as exogenous drug induced responses were recorded and analysed. The results of the study showed that the varicocele significantly inhibited the first phase of biphasic response of vas deferens in the ipsilateral side. However the correction of varicocele, free from surgical technique, ameliorated the affected first phase of EFS induced biphasic response in the ipsilateral side. The results of this study suggest that varicocele can be the reason of male infertility by not only causing testicular and/or epididymal damages but also triggering vas deferens motility defects. The correction of varicocele free from surgical technique may reverse the damaging of the vas deferens. Therefore when indicated surgical correction of varicocele is essential. It seems that varicocele surgery does not only prevent late term testicular and/or epididymal damages but also avoids vas deferens motility defects.

P53 EXPRESSION AND APOPTOSIS IN VARICOCELE IN THE RAT TESTIS

We investigated the presence of p53 expression and apoptosis in an experimental model of varicocele in the rat. A total of 30 adult male Sprague-Dawley rats were studied in 3 groups, namely group 1-13 with varicocele, group 2-9 with sham operation and group 3-8 controls. All animals underwent orchiectomy after 30 days and sections of the left testis from each animal were studied. TUNEL was used to detect apoptotic germ cells and an apoptotic index (AI) was calculated for each testis specimen. p53 expression was determined immunohistochemically and graded according to the proportion of cells stained in each specimen. AI values in group 1 testes were 0% to 0.51% (mean 0.06% +/- 0.13%) and the corresponding range in group 2 was 0.04% to 0.69% (mean 0.25 +/- 0.21%). No group 3 control specimens showed TUNEL staining (AI 0% in all). Although the mean AI in group 1 was higher than that in controls, this difference was not statistically significant (p = 0.318). The mean AI value in group 2 was significantly higher than in the control group (p = 0.021) but it was comparable to that in group 1 (p = 0.099). Seven (53.8%) and 2 (22.2%) specimens in groups 1 and 2, respectively, showed p53 staining in germ cell nuclei. None of the specimens in group 3 showed nuclear p53 expression. The grade of p53 expression in group 1 was significantly higher than that in groups 2 (p = 0.042) and 3 (p = 0.011). Expression grades in groups 2 and 3 were not significantly different (p = 0.426). To our knowledge p53 expression that may reflect abnormal spermatogenesis has been documented for the first time in an experimental model of varicocele in the rat testis. In contrast to other investigations, no link between apoptosis and varicocele was detected.