Melatonin therapy adds extra benefit to varicecelectomy in terms of sperm parameters,hormonal profile and total antioxidant capacity: A placebo‐controlled,double‐blind trial

In this study, we aimed at investigating the impact of melatonin supplementation on semen parameters, hormonal profile and total antioxidant capacity after varicocelectomy. Infertile male patients who were diagnosed with varicocele and underwent subinguinal varicocelectomy were included in the study. After performing subinguinal varicocelectomy, the patients were randomised into two groups: 27 receiving melatonin for 3 months and 27 as the placebo‐controlled group receiving placebo for 3 months. The pre‐operative parameters of semen analyses, hormonal profile and seminal oxidative stress status of both groups were compared with those of post‐operative parameters. There were statistically significant improvements in post‐operative parameters of semen analyses (sperm concentration, motility and proportions of normally formed spermatozoa), peripheral blood inhibin B and total antioxidant capacity in melatonin group compared with placebo group. In conclusion, melatonin therapy adds extra benefit to varicecelectomy in terms of sperm parameters, peripheral blood inhibin B and total antioxidant capacity; however, further studies including large number of samples are needed to make a proper decision on melatonin supplementation after varicocelectomy.     

VARICOCELE IS ASSOCIATED WITH ELEVATED SPERMATOZOAL REACTIVE OXYGEN SPECIES PRODUCTION AND DIMINISHED SEMINAL PLASMA ANTIOXIDANT CAPACITY

PURPOSE: Because varicocele is seen often in infertile men and oxidative stress has been implicated in sperm dysfunction, we assessed spermatozoal reactive oxygen species and seminal total antioxidant capacity in men with and without varicocele. MATERIALS AND METHODS: Levels of reactive oxygen species and total antioxidant capacity were measured in the semen of 21 infertile men with varicocele, 15 men with incidential varicocele and 17 normal donors without varicocele (controls). Men with leukocytospermia (more than 1 x 10(6) white blood cells per ml.) were excluded from study. Reactive oxygen species were measured in washed spermatozoa with a luminol dependent chemiluminescence assay. Total seminal antioxidant capacity was measured with an enhanced chemiluminescence assay, and the results were expressed as trolox equivalents. Sperm characteristics were assessed with a computer assisted semen analyzer, and sperm morphology was assessed using World Health Organization and Kruger's strict criteria. RESULTS: Patients with varicocele had significantly higher reactive oxygen species levels than controls (p = 0.02). Reactive oxygen species levels did not differ significantly between infertile and men with incidental varicocele. Total antioxidant levels were significantly lower among men with varicocele (p = 0.02) and those with incidental varicocele compared to controls (p = 0.05). Reactive oxygen species and total antioxidant capacity levels did not correlate in any group. CONCLUSIONS: Our results suggest that elevated reactive oxygen species and depressed total antioxidant capacity levels are associated with varicocele. These changes may be related to functional sperm abnormalities and infertility seen commonly in these patients. These findings support a possible rationale for controlled clinical trials of antioxidant supplementation in infertile men with varicocele.     

Pathophysiology and treatment options of varicocele: An overview

Varicocele is the most common correctable cause for male infertility, but not all men with varicocele are affected equally by this condition. The pathophysiology of varicocele-induced fertility remains ill-defined. While evidence suggests that oxidative stress remains a central factor, other mechanisms likely include scrotal hyperthermia, reflux of metabolites, hypoxia and cadmium accumulation. Microsurgical varicocelectomy remains the gold standard treatment option for infertile men with a clinically palpable varicocele and abnormal semen parameters. Newer evidence suggests a potential role for antioxidant supplementation and a meaningful role of varicocelectomy for patients destined for ART to improve pregnancy outcomes.     

Are antioxidants a viable treatment option for male infertility?

Oxidative stress is caused by an imbalance between ROS and antioxidants, which plays a significant role in the pathophysiology of many human diseases. There is extensive evidence highlighting the role of oxidative stress in male infertility due to elevated levels of sperm DNA fragmentation and abnormal semen parameters. The use of antioxidants is a potential therapeutic option to reduce ROS and improve semen quality. The appeal is that antioxidants can be easily obtained over the counter and are considered all-natural and therefore healthy. The hypothesis has been that by decreasing oxidative stress, antioxidants may be used for the treatment of male infertility. While initial studies of antioxidant supplementation suggested a beneficial role in the management of male subfertility, additional research has questioned the benefit of these therapies. The focus of this article is to present recent evidence assessing the viability of antioxidant therapy in the treatment of male infertility.     

4977‐bp mitochondrial DNA deletion in infertile patients with varicocele

Varicocele is the abnormal inflexion and distension of veins of the pampiniform plexus within spermatic cord and is one of the amendable causes of male infertility. It can increase reactive oxygen species (ROS) production in semen and cause oxidative stress. The purpose of this study was to analyse spermatozoa mtDNA 4977‐bp deletion in infertile men with varicocele. To detect 4977‐bp deletion in spermatozoa mtDNA, semen samples of 60 infertile patients with clinical varicocele and 90 normal men from northern Iran were prepared. After extraction of spermatozoa total DNA, Gap polymerase chain reaction (Gap PCR) was performed. 4977‐bp deletion was observed in 81.66% of patients with varicocele, while approximately 15.55% of controls had this deletion. As spermatozoa from patients with varicocele had a high frequency of occurrence of 4977‐bp deletion in mtDNA [OR = 24.18, 95% confidence interval (CI) = 10.15–57.57, P < 0.0001], varicocele may induce mtDNA deletion in spermatozoa and cause infertility in north Iranian men. However, to determine the relation between sperm mtDNA 4977‐bp deletion and varicocele‐induced infertility, larger population‐based studies are needed. It is concluded that there is an association between sperm mtDNA 4977‐bp deletion and varicocele‐induced infertility in the population studied.     

Long-term NaHS administration reduces oxidative stress and apoptosis in a rat model of left-side varicocele

The main aim of this study was to assay the testicular H₂S levels in the varicocele rat model and then to investigate the protective effects of NaHS on morphometric changes, sperm parameters, oxidative stress and apoptosis markers in rat's testis. D,L-propargylglycine (PAG) was administrated to show the effects of cystathionine γ-lyase enzyme (CSE) inhibition in the varicocele. Rats were assigned to four groups: (a) Sham, (b) varicocele, (c) varicocele + PAG and (d) varicocele + NaHS. Animals in varicocele + NaHS group received 30 µmol/L NaHS in drinking water for 56 days. In the varicocele + PAG group, animals received PAG 19 mg/kg twice a week. Morphometric assessment, oxidative stress markers, testicular H₂S levels, sperm parameters, TUNEL assay and expression of Bax/Bcl2 were evaluated at the end of experiment. Testicular H₂S levels were significantly decreased in varicocele group. NaHS significantly improved sperm parameters, morphometric characteristics and oxidative stress compared to varicocele group. Oxidative stress status deteriorated in the PAG group compared to the varicocele group. This study showed that a low testicular H₂S level might play a critical role in male infertility. Thus, NaHS administration may be a promising treatment strategy for male infertility in varicocele. In addition, CSE may not be the only important enzyme in testicular H₂S production.     

Oxidative stress before and after surgery in benign prostatic hyperplasia patients

The aim was to assess oxidative stress in benign prostatic hyperplasia patients and also to evaluate the effect of operation in late (60 days) post‐operative period. This study was conducted with 16 patients with benign prostatic hyperplasia and 16 healthy subjects. Serum malondialdehyde, blood 8‐hydroxy‐2′‐deoxyguanosine/deoxyguanosine, erythrocyte superoxide dismutase, serum total coenzyme Q₁₀ and coenzyme Q₁₀ levels were measured. Independent samples t test was used to analyse the differences between control group and patients, while paired t test was used to analyse the differences between pre‐operative and post‐operative periods. Malondialdehyde and total coenzyme Q₁₀ levels were lower in patients, while 8‐hydroxy‐2′‐deoxyguanosine/deoxyguanosine level was increased. However, superoxide dismutase activity and coenzyme Q₁₀ levels did not differ. After 60 days of operation, 8‐hydroxy‐2′‐deoxyguanosine/deoxyguanosine and superoxide dismutase activity decreased, while total coenzyme Q₁₀ level increased. However, malondialdehyde and coenzyme Q₁₀ levels were not affected. The international prostate symptom scores of the patients were also decreased after the operation. The results suggest that blood 8‐hydroxy‐2′‐deoxyguanosine/deoxyguanosine level may reflect the oxidative stress better than the malondialdehyde level, and surgical operation attenuates the oxidative stress in late post‐operative period in benign prostatic hyperplasia patients.     

The effect of coenzyme Q10 supplementation on partner pregnancy rate in infertile men with idiopathic oligoasthenoteratozoospermia: an open-label prospective study

Objective

It has been shown that coenzyme Q₁₀ (CoQ₁₀) supplementation in men with idiopathic oligoasthenoteratozoospermia (OAT) results in improved semen parameters. In present study, we evaluated the effects of coenzyme CoQ₁₀ supplementation on semen parameters and pregnancy rates in infertile men with idiopathic OAT.

Patients and methods

Two hundred and eighty-seven infertile men with idiopathic OAT were recruited in this study. These patients were treated with CoQ₁₀ 300?mg orally twice daily for 12?months. Two semen analyses and determination of resting levels of sex hormones were done in all participants. Patients were followed up for another 12?months after CoQ₁₀ discontinuation.

Results

Mean sperm concentration, sperm progressive motility, and sperm with normal morphology improved significantly after 12-month CoQ₁₀ therapy by 113.7, 104.8, and 78.9%, respectively (all Ps?< 0.05). The overall pregnancy rate was 34.1% within a mean of 8.4?±?4.7?months.

Conclusions

CoQ₁₀ supplementation improves semen quality with beneficial effect on pregnancy rate.     

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.     

Assessment of seminal granulysin in infertile men with varicocele

Varicocele has a common association with male infertility, but its exact role is still debated. Apoptosis has been suggested as one of the mechanisms of varicocele‐associated infertility. Granulysin is a molecule that plays a role in apoptosis with no previous study about its role in male infertility. This case‐controlled study aimed to assess seminal plasma granulysin level in infertile patients with varicocele. This study involved 90 men that were allocated into fertile normozoospermic men (n = 20), infertile men without varicocele (n = 30) and infertile men with varicocele (n = 40). These men were subjected to history taking, clinical examination, semen analysis and estimation of seminal granulysin. In general, seminal granulysin level was significantly elevated in infertile men compared with fertile men. Infertile men with varicocele showed significantly higher seminal granulysin compared with infertile men without varicocele, in bilateral varicocele cases and in grade III varicocele. Seminal granulysin level was negatively correlated with sperm concentration, sperm motility, sperm normal forms percentage and testicular volumes. It is concluded that increased seminal granulysin has a negative impact on spermatogenesis in infertile men in general and in infertile men associated with varicocele in particular.     

Effect of varicocele repair on sperm DNA fragmentation: a review

Varicocele, the leading cause of male infertility, can impair sperm quality and fertility via various oxidative stress mechanisms. An imbalance between excessive reactive oxygen species production and antioxidant protection causes alterations in nuclear and mitochondrial sperm DNA, thus rendering a subset of varicocele men less fertile. In particular, sperm DNA fragmentation is usually elevated in men with clinical varicocele in both abnormal and normal semen parameters by the current World Health Organization criteria. In this review, we discuss the evidence concerning the association between varicocele, oxidative stress, and SDF, and the possible mechanisms involved in infertility. Furthermore, we summarize the role of varicocele repair as a means of alleviating SDF and improving fertility. Lastly, we critically appraise the evidence-based algorithm recently issued by the Society for Translational Medicine aimed at guiding urologists on the use of SDF testing in men with varicocele seeking fertility. Current evidence based on careful review of published studies confirms the effectiveness of varicocelectomy as a means of both reducing oxidatively induced sperm DNA damage and potentially improving fertility. Varicocele repair should be offered as part of treatment option for male partners of infertile couples presenting with palpable varicoceles.     

Antioxidant effects of penicillamine against in vitro-induced oxidative stress in human spermatozoa

Oxidative stress contributes importantly to the aetiology of male infertility, impairing sperm function. The protective effect of antioxidants on seminal parameters has been established, and the antioxidant penicillamine has shown beneficial effects; however, its protective effect on human spermatozoa exposed to oxidative stress has not been reported. The objective of this work was to evaluate the effect of penicillamine on human spermatozoa exposed in vitro to oxidative stress. First, the effect of penicillamine on spermatozoa from normozoospermic donors was evaluated. Then, the effect of penicillamine on spermatozoa exposed to oxidative stress induced separately by ionomycin and hydrogen peroxide (H₂O₂) was analysed. An untreated control and a control treated only with the oxidative stress inducer were included. Reactive oxygen species (ROS) levels, viability, mitochondrial membrane potential (MMP) and motility were analysed. The results showed that penicillamine, added to the incubation medium, decreased the ROS levels induced by ionomycin and H₂O₂, and this effect was associated with better preservation of MMP, motility, and ATP levels. These results highlight the potential advantages of penicillamine supplementation of sperm culture medium, especially for semen samples with high ROS levels and also in circumstances where laboratory handling can cause an increase in ROS production.     

The effects of dietary lycopene supplementation on human seminal plasma

OBJECTIVE: To investigate whether lycopene levels in blood and seminal plasma increase after dietary supplementation with a natural source of the compound, and whether any potential increase of lycopene levels in semen translates into increased free-radical trapping capacity in the seminal plasma. METHODS: Reactive oxygen species are detrimental to the health and function of spermatozoa. Semen contains enzymatic and non-enzymatic defence mechanisms to combat such species, and lycopene, a dietary antioxidant, forms part of the non-enzymatic arm. Immuno-infertile men have significantly lower levels of lycopene in their semen, and oral lycopene therapy can improve various seminal variables in idiopathic infertility. Whether this improvement is a direct consequence of increased lycopene levels in semen, resulting in an increased radical scavenging ability, remains unknown. Blood and seminal lycopene levels were measured in healthy volunteers, using high-performance liquid chromatography, before and after a period of dietary supplementation. The antioxidant capacity of seminal plasma was also assayed to determine if supplementation results in a measurable increase in seminal radical scavenging ability. RESULTS: There were statistically significant increases in blood and seminal plasma lycopene levels after dietary supplementation. The increase in seminal and blood lycopene levels showed a strong positive correlation (r = 0.84, P < 0.05). There was no measurable increase in the total radical scavenging capacity of semen. CONCLUSION: This study confirms the presence of lycopene in human semen, the levels of which can be significantly increased after dietary supplementation with a natural source of lycopene. Further studies to establish whether this would also be the case in infertile men, with possible associated improvements in their seminal quality, are warranted.     

Oxidative stress status and sperm DNA fragmentation in fertile and infertile men

Evidence suggests that disturbing the balance between reactive oxygen species levels and antioxidant contents in seminal plasma leads to oxidative stress resulting in male infertility. This study was carried out to identifying clinical significance of seminal oxidative stress and sperm DNA fragmentation in treatment strategies of male infertility in southwest Iran. Sperm parameters, lipid peroxidation and activity of antioxidant enzymes were assessed in fertile (n = 105) and infertile (n = 112) men. Malondialdehyde (MDA) levels in seminal plasma were found to be higher significantly (p < .001) in patients. Superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities in seminal plasma were significantly (p < .001) lower in infertile men. Significant negative correlations were observed between MDA levels and sperm motility and normal morphology. Spermatozoa with fragmented DNA were higher (p < .001) in infertile men and significantly correlated with MDA levels and SOD and GPx activities. MDA of 4.2 nmol/ml, SOD of 4.89 U/ml and GPx of 329.6 mU/ml were optimum cut‐off limits to discriminate infertile patients from fertile men. The results show the leading role of oxidative stress in aetiology of male infertility in southwest Iran and indicate that evaluation of seminal antioxidant status and DNA integrity can be helpful in men attending infertility clinics during fertility assessment.     

Oxidative stress: a common factor in testicular dysfunction

Oxidative stress results from the production of oxygen radicals in excess of the antioxidant capacity of the stressed tissue. Many conditions or events associated with male infertility are inducers of oxidative stress. X-irradiation, for example, or exposure to environmental toxicants and the physical conditions of varicocele and cryptorchidism have been demonstrated to increase testicular oxidative stress, which leads to an increase in germ cell apoptosis and subsequent hypospermatogenesis. Such stress conditions can cause changes in the dynamics of testicular microvascular blood flow, endocrine signaling, and germ cell apoptosis. Testicular oxidative stress appears to be a common feature in much of what underlies male infertility, which suggests that there may be benefits to developing better antioxidant therapies for relevant cases of hypospermatogenesis.     

Blood and semen paraoxonase—arylesterase activities in normozoospermic and azoospermic men

Paraoxonase and arylesterase enzymes are corner stones of antioxidant defence. We aimed to compare azoospermic infertile men and normozoospermic individuals with respect to total antioxidant status (TAS), total oxidant status (TOS), oxidative stress index (OSI), paraoxonase and arylesterase levels in the blood and seminal plasma. Two‐hundred consecutive infertility patients and voluntarily participated were included. In the normozoospermic group, TAS, PON, arylesterase values were statistically significantly higher when compared with those in the azoospermic group, while lower TOS and OSI levels were observed in the blood and seminal plasma of azoospermic group. In the semen analyses of normozoospermic group, the correlation between semen volume, sperm concentration, sperm motility and morphology and TAS, TOS, OSI, PON and arylesterase values was examined. A negative correlation was determined between semen volume and OSI. Levels of serum oxidative parameters were higher in the azoospermic group relative to normozoospermic group, but antioxidant parameters were lower than those of the normozoospermic group. Oxidative stress performs an essential role in the aetiology of male infertility by negatively influencing sperm quality and function. Assessment of blood and seminal plasma oxidative profiles might be an important tool to better evaluation of sperm reproductive capacity and functional competence.     

The assessment of oxidative stress in infertile patients with varicocele

OBJECTIVES

To assess oxidative stress markers, antioxidant capacity and cytokines in seminal plasma from infertile patients with varicocele, and to investigate seminal oxidative status and sperm DNA damage after varicocelectomy.

PATIENTS, SUBJECTS AND METHODS

The records were retrospectively evaluated for 28 azoospermic, 30 oligospermic (15 with varicocele and 15 without) and 30 patients with normal semen characteristics (15 with varicocele and 15 without). The mean (sd ) age of the men was 32.4 (5.6) years; all men with varicocele had a unilateral or bilateral microsurgical subinguinal varicocelectomy. The level of nitric oxide (NO), 8‐hydroxy‐2′‐deoxyguanosine (8‐OHdG), hexanoyl‐lysine (HEL), superoxide dismutase (SOD) activity, interleukin (IL)‐6, IL‐8 and tumour necrosis factor‐α in seminal plasma were measured. In addition, sperm DNA fragmentation was analysed before and 6 months after varicocelectomy.

RESULTS

Azoospermic and oligospermic patients had a significantly higher HEL concentration and SOD activity in seminal plasma; those with varicocele had a significantly higher NO, HEL, and SOD activity in seminal plasma. There was a significant increase in sperm concentration and reduction in NO, HEL, 8‐OHdG level and SOD activity after varicocelectomy. Oligospermic patients with varicocele had a significantly higher IL‐6 level in seminal plasma, and there was a significant reduction after varicocelectomy. The percentage of apoptosis‐positive sperm decreased significantly after varicocelectomy.

CONCLUSIONS

This study indicates that the seminal plasma of patients with varicocele is under excessive oxidative stress, and partly even in patients with normospermia, and that varicocelectomy reduces oxidative stress in seminal plasma and ameliorates sperm DNA damage.     

Supplementing post-wash asthenozoospermic human spermatozoa with coenzyme Q10 for 1 hr in vitro improves sperm motility,but not oxidative stress

We investigated the effect of supplementing post-wash asthenozoospermic spermatozoa with coenzyme Q10 (CoQ10) in vitro, which may reduce oxidative stress and improve sperm motility. Semen samples were collected from 39 men with asthenozoospermia, and their spermatozoa were isolated by two-layer Percoll density-gradient centrifugation. Kinetic parameters of the isolated spermatozoa (baseline before intervention) were determined immediately by computer-aided semen analysis. Total anti-oxidant capacity and protein carbonyl levels, as markers of oxidative stress, were also measured in the baseline spermatozoa. The baseline spermatozoa suspension was divided equally into two portions, one for CoQ10 supplementation (50 µg/ml for 1 hr) and the other as an un-supplemented vehicle control. The total motility of the CoQ10-supplemented spermatozoa was significantly higher than in the control (p = .009) and progressive motility tended to be higher (p = .053). Immotile sperm concentration in the CoQ10-supplemented spermatozoa was significantly lower than in both the baseline (p = .026) and control (p = .009). Total anti-oxidant capacity and protein carbonyl levels between the baseline, CoQ10-supplemented and control spermatozoa were not significantly different. Our data suggest that CoQ10 treatment reactivated sperm motility. We propose short-term supplementation of post-wash asthenozoospermic spermatozoa with CoQ10 before intrauterine insemination.     

Evaluation of plasma oxidative stress,with or without antioxidant supplementation,in superficial partial thickness burn patients: a pilot study

Background: Oxidative stress is one of the main causes of pathophysiological alterations observed during burn injury. The present pilot study aimed to determine whether a specific oral antioxidant supplementation could in any way influence free radical blood values in patients affected by superficial partial thickness burns.

Materials and methods: Plasma oxidants and plasma antioxidant capacity were analysed in 20 superficial partial thickness burn patients for a 2-week period; patients were randomly divided into two groups, one of which was supported with a specifically designed oral antioxidant formula (Squalene 100?mg, Vitamin C 30?mg, Coenzyme Q10 10?mg, Zinc 5?mg, Beta Carotene 3.6?mg, Bioflavonoids 30?mg, Selenium 55?mcg) administered daily, starting from the day of admission, for the whole study period.

Results: No significant differences were found in plasma oxidants and plasma antioxidant capacity between the two groups of patients.

Conclusions: These results did not reflect any significant benefits of an antioxidant oral supplementation at usual dosages when considering oxidative plasmatic values of superficial partial thickness burn patients.     

Alpha- and gamma-tocopherol levels in human semen and their potential functional implications

Vitamin E is a potent antioxidant that has been considered involved in fertility, but studies have mostly focused on α-tocopherol. Our study aimed at measuring, by an isotope dilution gas chromatography–mass spectrometry method, α- and γ-tocopherol concentration in human semen in a large and well-characterised population (134 men with different semen parameters and in varicocele patients), as well as their potential role in male fertility. We carried out freeze/thaw experiments in 15 samples with the two isomers in the cryoprotective medium. Moreover, our study included 10 subjects supplemented in vivo with α-tocopherol for 90 days. In seminal plasma, γ-tocopherol concentration was significantly lower in the varicocele group than in the normozoospermic group. We observed that γ-tocopherol, supplemented to cryopreservation medium, induced a higher post-thaw human sperm viability and motility than α-tocopherol. The results of in vivo α-tocopherol supplementation showed a decrease in γ-tocopherol concentration with increasing α-tocopherol level in blood. This is the first report related to γ-tocopherol distribution in human semen analysed by gas chromatography–mass spectrometry. γ-tocopherol would not seem to be related to semen parameters but to cellular oxidative condition. This tocopherol may contribute to human health in a yet unexplored way.