SummaryBackgroundInfertility is an important worldwide problem which affects 10–15% of couples globally. Altered NO production has also been implicated in the pathogenesis of the male infertility. The present study was designed to evaluate the changes in the activity of NO-synthase (NOS) and arginase in spermatozoa of patients with infertility.MethodsThe total NOS, Ca2+-dependent constitutive (cNOS) and Ca2+-independent inducible (iNOS) activity and arginase activity were assessed in sperm cells of patients with different forms of pathospermia.ResultsWe found a significant increase in iNOS activity, but significantly decreased cNOS and arginase activity in sperm cells of infertile men vs fertile, normozoospermic men (p<0.001). The arginase/NOS ratio significantly decreased compared to control group. The iNOS/cNOS ratio was drastically increased in patients with decreased fertility potential indicating predominance of iNOS. Men with leuko cytospermia were distinguished to have the most express iNOS activity.ConclusionsThese observations provide evidence for a disturbed balance between the L-arginine metabolic pathways in sperm cells of infertile men. This imbalance includes the considerable activation of the inducible isoform of NO-synthase accompanied by significant inhibition of its constitutive isoform which indicates disturbances in NO production. In patients with decreased fertility potential the arginase/NOS was shifted towards predominance of iNOS-derived NO production.
SummaryBackgroundCoronary heart disease is the leading cause of death and disability worldwide. Hypertension is a major independent risk factor for the development of CHD. Abnormalities in NO generation or activity have been proposed as a major mechanism of CHD. The purpose of this article is to determine the activity of eNOS and iNOS in patients with isolated CHD and CHD associated with HT of different age groups.MethodsFifty patients with isolated CHD and 42 patients with CHD associated with HT were enrolled in this study. NOS activity was determined by nitrite anion formed in the reaction.ResultsA statistically significant increase in iNOS activity is observed in elderly donors. In patients with isolated coronary heart disease cNOS activity is statistically significantly reduced with respect to the control group. The reduction of enzymatic activity of cNOS is more expressed in elderly patients than in middle-aged patients with coronary heart disease. Alterations in eNOS activity are more expressed in patients with coronary heart disease associated with hypertension than in patients with isolated coronary heart disease. Against the background of cNOS inhibition in the patients, a sharp increase in iNOS activity is observed.ConclusionsIt has been shown that disturbance of endothelial function in patients with coronary heart disease associated with hypertension is characterized by reduced endothelial NO synthesis by cNOS and increased systemic NO synthesis due to increased iNOS activity. It has been found that the lack of endothelial NO and hyperproduction of »harmful« NO by iNOS are more expressed in elderly patients.
Background: Oxidative stress is considered as one of the causes of male subfertility or infertility. Among antioxidant enzymes, the crucial role belongs to glutathione S -transferases (GSTs). Data on the biological role of GSTs in the defense mechanisms of sperm cells in fertile and infertile men are limited. Aim: The aim of this study was to demonstrate the functional role of GSTs in sperm cells on the model of H 2 O 2 -induced stress on human ejaculated spermatozoa obtained from both normospermic and pathospermic patients. Subjects and Methods: We used a H 2 O 2 -induced stress on human ejaculated spermatozoa obtained from both normospermic and pathospermic patients. Results: Here, we report the effect of GST inhibitor ethacrynic acid on sperm motility and viability. Pharmacological inhibition of sperm GSTs activity leads to spermal membrane damage and rejected in the loss of motility and decrease of viability. For similar treatment conditions, thiobarbituric acid reactive substance (TBARS) levels increased significantly leading to decrease in sperm motility and viability. It is suggested that these functional impairments are related to the intensification of lipid peroxidation as expressed by TBARS levels in spermal membranes after GST inhibitor treatment. Conclusion: This study provides evidence that sperm GSTs are important in the defense mechanism against oxidative stress. Evaluation of GSTs activity in sperm cells of infertile men can be helpful in fertility assessment and for the evaluation of treatment by antioxidants.
It is believed that the most common causes of male infertility are impairment of spermatogenesis and sperm functions. Glutathione S-transferases (EC 2.5.1.18) play an important role in sperm physiology, specifically in antioxidant protection against oxidative damage. The catalase decomposition of lipid hydro-peroxides forms as a result of oxidative stress. We used a model of superoxide anion-generating system Fe3+/ascorbate or H2O2-induced stress to study the activity of glutathione s transferase in human ejaculated spermatozoa from patients with pathospermia and products of lipid peroxidation (TBARS) as a marker of oxidative stress. In the present study, dose dependent increase in the level of lipid peroxidation was observed for treatment with Fe3+/ascorbate or H2O2. The TBARS level was higher for sperm cells incubated with superoxide anion-generating system Fe3+/ascorbate than for H2O2. GSTs activity increased in spermatozoa treated with increasing concentration of superoxide anion-generating system Fe3+/ascorbate and H2O. We found that both Fe3+/ascorbate and H2O2 displayed similar inhibitory effects on sperm GSTs activity, however H2O2 at low concentrations activated enzyme activity only in normozoospermic samples, which can be explained as a defence response to oxidative stress. The time course of incubation with 100 μM H2O2 showed a sharp decrease in the enzyme activity during the first 5 min of incubation for both normozoospermic and pathozoospermic men. Preincubation of spermatozoa with GSH completely prevented the ROS-induced inhibition on GSTs only in normozoospermic samples. On the other hand, in pathospermic samples protectory effect of GSH was observed only against non-radical (H2O) radical, but not against radical (superoxide anion-generating system Fe3+/ascorbate) species. The results of our study showed higher oxygen-free radical production, evidenced by increased TBARS level in spermatozoa obtained from infertile men than normozoospermic men. The inhibitory effect of the radical (superoxide anion-generating system Fe3+/ascorbate) species on sperm GSTs activity and products of lipid peroxidation in sperm cells of fertile and infertile men were more expressed compared to non-radical (H2O) species. Our results indicate that estimation of sperm GSTs enzyme assays can be used as a bioindicator for impaired male fertility. The obtained results argue for a biological role of sperm GSTs in susceptibility of spermatozoa to oxidative damage and maintaining sperm antioxidant status.
Background. Male infertility is one of the most serious medical and social problems. Idiopathic infertility accounts for about 30 % of cases of infertile men. Rheumatoid arthritis is associated with a decreased fertility potential. The aim of the present work was to determine the lipid peroxidation level and the activity of antioxidant enzymes such as glutathione peroxidase and glutathione reductase in seminal plasma and blood plasma of infertile men with idiopathic infertility and concomitant autoimmune joint pathology (rheumatoid arthritis). Materials and Methods. 45 infertile men aged 22–48 were examined. They were divided into 2 groups: first group – 23 somatically healthy patients with idiopathic infertility; second group – 22 infertile men with rheumatoid arthritis. The control group consisted of 27 males with normal semen profile according to the WHO criteria and confirmed parenthood. The concentration of thiobarbituric acid reactive substance and activity of antioxidant enzymes were measured in the blood and seminal plasma. Results. When analyzing the seminal fluid, we found that the TBARS content was 4-fold greater in infertile men with autoimmune pathology compared to fertile men (p <0.001), whereas, in patients with idiopathic infertility its level was within the normal range. The activation of lipid peroxidation in infertile men with idiopathic infertility and in combination with rheumatoid arthritis was accompanied by a statistically significant decrease in the activity of enzymes of glutathione antioxidant system. It should be noted that more pronounced disorders of lipid peroxidation and antioxidant enzymes activity were found in seminal plasma compared to blood plasma. Conclusions. (1) An increased lipid peroxidation was observed in seminal and blood plasma of infertile men in combination with rheumatoid arthritis compared to normospermic men, whereas no differences were observed between men with idiopathic infertility and fertile men; (2) An impaired antioxidant status was observed in seminal and blood plasma of both men with idiopathic infertility and infertile men in combination with rheumatoid arthritis compared to normospermic men; (3) infertile men in combination with rheumatoid arthritis showed a significantly higher lipid peroxidation levels compared to men with idiopathic infertility, whereas no differences were observed in GPx and GR activity between groups.
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