УДК 616.697:616.72-002.77-092:612.015.11]-07

According to modern ideas, the development of pathological processes in the body, in particular relat-ed to infertility, is accompanied by a violation of the mechanisms of antioxidant protection of cells. Hyperproduction of reactive oxygen species (ROS) is found in many pathological conditions, both related to the reproductive system and not directly related to it. Excessive ROS production, is thought, to be caused by an imbalance between prooxidants and the antioxidant system and causes damage to the sperm membrane, chromosome DNA, and initiates sperm apoptosis that leading to infertility.The aim – to study of the functional state of the pro- / antioxidant system in the seminal fluid and serum of infertile men with concomitant pathology.Material  and  Methods. The studies were performed on the serum and seminal fluid of infertile men aged 22–48 years with diagnosed idiopathic infertility (n=23) and rheumatoid arthritis (n=22). The control group consisted of 27 men of the same age. The ejaculate was analyzed according to WHO recommendations (2010). Determination of malonic dialdehyde (MDA) concentration was performed based on its interaction with 2-thiobarbituric acid. The state of the antioxidant system of semen and blood serum was concluded based on determining the total antioxidant activity by enzyme-linked immunosorbent assay using a commercial test system manufactured by Elabscience (ELISA, USA). Results. It was found that in patients with autoimmune pathology, the concentration of MDA in the serum exceed-ed the control group by 2.25 times, and in idiopathic infertility – by 2.16 times. In semen, the content of MDA in men with infertility due to autoimmune pathology is likely to exceed 4 times the control values. In patients with idiopathic infer-tility, the content of MDA in the seminal fluid was within normal limits. Against the background of intensification of lipid peroxidation processes, the overall antioxidant activity decreases. At autoimmune pathology it decreases in 8.1 times.Conclusion. Oxidative stress is one of the immunopathogenetic mechanisms of development of male infertility against the background of autoimmune pathology and idiopathic infertility.

УДК 616.697-07-091.8

Питома вага чоловіків з непліддям постійно зростає як в Україні, так і в усьому світі. За від-сутності достовірних етіологічних чинників виникнення непліддя його ідентифікують як ідіопатичне. Воно трапляється у 25–30 % чоловіків. Щодо патохімічних досліджень, то припускається, що в багатьох випадках ідіопатичне непліддя чоловіків спричинене оксидативним стресом. Однак також відомо, що важлива регуляторна роль у підтриманні клітинного гомеостазу належить оксиду азоту як вторинному месенджеру – універсальному регулятору практично всіх фізіологічних функцій організму. Мета дослідження – з’ясувати функціональну активність ензимів аргіназа/NO-синтазної системи та виявити нові потенційні маркери ідіопатичного непліддя чоловіків. Методи дослідження. Дослідження проводили на сім’яній плазмі 33 інфертильних чоловіків з ідіопатичним непліддям віком 22–48 років. До контрольної групи ввійшли 27 фертильних практично здорових чоловіків того ж віку. В сім’яній плазмі визначали активність таких ензимів, як аргіназа, конститутивна й індуцибельна ізоформи NO-синтази (сNOS, іNOS), а також концентрацію нітрат- та нітрит-аніонів (NO3-, NO2-)і малонового діальдегіду (МДА).Результати й обговорення. При ідіопатичному неплідді чоловіків активність аргінази в сім’яній плазмі зменшилася в 1,6 раза. Одночасно знизилась активність сNOS в 1,3 раза і зросла активність іNOS у 24,6 раза. Виявлене зменшення співвідношення аргіназа/NOS у 3,1 раза засвідчило зниження активності неокисного, аргіназного метаболізму, який конкурує з окисним NO-синтазним метаболізмом L-аргініну. Зниження в сім’яній плазмі співвідношення активності аргiназа/іNOS у 25,7 раза може бути важливим показником розвитку ідіопатичного непліддя. Під час оцінювання інтенсивності функцioнування NO-син-тазного шляху метаболізму L-аргініну при ідіопатичному неплідді відзначено вірогідне зростання в сім’я-ній плазмі концентрації NO3- в 1,4 раза та зниження NO2- в 1,5 раза. Виявлено пряму кореляційну залежність між концентрацією МДА і стабільних метаболітів оксиду азоту – NO3-. Продемонстрований прямий кореляційний зв’язок між активністю іNOS та концентрацією МДА в сім’яній плазмі (r=0,87) свідчить про те, що при ідіопатичному неплідді іNOS може активувати процеси пероксидації ліпідів. Висновки. Розвиток ідіопатичного непліддя чоловіків супроводжується змінами активності ензимів аргіназа/NO-синтазної системи, зокрема зниженням активності аргінази та сNOS і зростанням активності іNOS, що свідчать про домінування окисного шляху метаболізму L-аргініну. Зменшення в сім’яній плазмі співвідношення активності аргiназа/іNOS може бути важливим показником розвитку ідіопатичного непліддя.

УДК 616.697-07-091.8

The most difficult form of male infertility to treat is azoospermia. It is identified as a complete absence of sperm in the ejaculate. Depending on the nature and causes of impairedspermatogenesis, azoospermia is divided into obstructive  (excretory,  OA)  and  non-obstructive  (secretory,  NOA).  Testicular  biopsy  is  the  most  effective  method  of diagnosis and a component of possible treatment for azoospermia. It is the only objective method of differential diagnosis between non-obstructive and obstructive forms of azoospermia.The aim:histological analysis of testicular biopsies of men with various forms of azoospermia. Мaterials and Methods.78 infertile men with azoospermia were examined. They were divided into two groups: the first group, men with NOA (n = 28); the second group, men with OA (n = 50). The biopsy was preceded by a manda-tory ultrasound  diagnosis  of  the  portal  system. The  biopsy was  performed  using  the  method  of  open  operative access. Biopsies were mostly taken from the more palpable testicle or from both testicles. Biopsies were fixed in buffered 10% formalin (pH 7.2). After 1 day, they were dehydrated in 70% ethanol and embedded in paraffin. For histological studies, sections with a thickness of 5 μm were stained with hematoxylin and eosin. Biopsies were evaluated in accordance with previously described methods.Results.Histological analysis of testicular biopsies from 28.7% of patients with a non-obstructive form of azo-ospermia showed swelling of the testicular stroma, destructive changes in testosterone-producing cells, disruption of the structure of the syncytial complexes of the spermatogenic epithelium, and the complete absence of the process of sper-matogenesis in individual tortuous seminiferous tubules, the absence of contacts between sustentocytes, and in erythro-cyte sludge in the lumen of vessels. 42.8% of patients had fibrosis of the testicular stroma, stroma swelling, thinning of the wall of convoluted seminiferous tubules, violation of the structure of the syncytial complexes of the spermatogenic epithelium, proliferation of the wall of the convoluted seminiferous tubules into their lumen, and infiltration of the testic-ular  stroma  with  lymphocytes. In  54.0%  of  patients  with  preserved  spermatogenesis  and an  obstructive  form of  azoo-spermia, it was possible to find a history of orchoepididymitis in the anamnesis; one patient (2.0%) underwent bilateral orchopexy  at  the age  of 5  years due  to  cryptorchidism;  6.0%  recalled the  trauma calculi  in  the anamnesis; and 38.0% denied any factors affecting fecundity in the anamnesis.Conclusions.The non-obstructive form of azoospermia is characterized by the following parameters: mostly a violation of the structure of the spermatogenic epithelium, a complete absence of the process of spermatogenesis in indi-vidual  convoluted  seminiferous  tubules,  a  violation  of  the  structure  of  the  hematotesticular  barrier,  and  a  violation  of blood microcirculation. The histological picture of preserved spermatogenesis is of the same type in 88.0% of patients with an obstructive form of azoospermia. In most tubules, a fixed number of cell rows is preserved, and cells of various stages of spermatogenesis are determined in them: spermatogonies, spermatocytes, a moderate number of spermatids.In the lumen of the tubules, exfoliated cells and a moderate number of spermatozoa are found.

Abstract. The mechanisms of formation of male infertility are increasingly becoming immune-dependent. Immunological isolation of the testes is provided by the anatomical blood-testis barrier and the special tolerance of the immune system to antigens expressed on male gametes. For a better understanding of the immunopathogenetic mechanisms of infertility, a study of the role of various immune factors is required.
The aim: to study the role of pro-inflammatory and anti-inflammatory cytokines in blood serum to clarify their role in spermatogenesis.
Materials and Methods: 45 infertile men aged 22-48 were examined. They were divided into 2 groups: first group – 22 men with a systemic autoimmune disease - rheumatoid arthritis; second group – 23 somatically healthy patients
with idiopathic infertility. The control group included 27 fertile healthy men aged 22-48 years. Determination of the
cytokines concentration in blood serum was carried out by the immunoenzymatic method. Student's t-test was used to compare the significant difference in mean values between groups. P < 0.05 was considered significant.
Results. Patients with autoimmune diseases (rheumatoid arthritis) had the highest rate of leukocytospermia, indicating a possible long-term inflammatory process. In patients with idiopathic infertility, oligozoospermia was diagnosed in 3 patients (13.04%), oligoasthenozoospermia in 7 patients (30.36%), asthenozoospermia in 8 patients (34.78%) and leucocytospermia in 5 patients (21.82%). In patients with rheumatoid arthritis, the concentration of IL-18 (cytokine of the IL-1 family) in the blood serum of infertile men was 1.36 times higher than that of fertile men. The level of IL-6 was 6 times higher, and the concentration of IFN-γ exceeded the norm by more than three times. The level of IL-10 was 9,4 times higher than in control group. Significant changes in the serum cytokine profile were recorded in men with idiopathic infertility. The level of pro-inflammatory cytokines increased statistically significantly: IL-18 – 1.45 times, IL6 – 2.85 times, IFN-γ – 2.65 times. Simultaneously, the level of anti-inflammatory cytokines increased: IL-10 – 3.0 times.
We also analyzed the ratio of serum levels of pro- and anti-inflammatory cytokines. Significant increase in ІL10/TNF-α ratio was recorded in men with idiopathic infertility and infertile men with rheumatoid arthritis. Specifically, in men with idiopathic infertility the ІL-10/TNF-α ratio was 3.3 times higher and in patients with rheumatoid arthritis the ІL-10/TNF-α ratio was 4.1 times higher than in control group. In patients with rheumatoid arthritis the ІL-10/ІFN-γ ratio
was in 3 times higher than in control group. Significant positive correlations were found in healthy fertile men: TGF-β1 – IFN-γ, IL-10 – IL-18 and IL-10 – TNF-α. In men of the control group, the synthesis of IL-18 and TNF-α was balanced by the production of IL-10. The synthesis of IFN-γ was by balanced by the production of TGF-β1.
Conclusions. The development of infertility associated with various accompanying pathologies is related with by changes in both systemic and local immune reactivity. The pro-inflammatory cytokine profile of blood serum and a decrease in the concentration of IL-1β are observed in patients with idiopathic infertility. The largest number of deviations of immune reactivity was found in infertile men with concomitant autoimmune diseases

УДК: 616.697-008.8-06:616.72-002-092:612.015.11]-07

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.