RESUMO
Utilizando bases de datos Medline, Science Direct, Endocrine Society y Sociedad Argentina de Andrología se realizó una revisión bibliográfica para estudiar e interpretar el efecto del estrés oxidativo (EO) en el proceso reproductivo. Los seres vivos que utilizan oxígeno para obtener energía son liberadores de especies reactivas de oxígeno (ERO). Existen situaciones andrológicas, factores medioambientales y compuestos químicos que incrementan las citoquinas proinflamatorias, generan EO alterando la regulación del proceso espermatogénico. Los espermatozoides son susceptibles al daño oxidativo, debido al elevado contenido de ácidos grasos poliinsaturados (AGPI) en su membrana y el escaso citoplasma que limita el contenido de enzimas antioxidantes. El EO induce daño peroxidativo en la membrana espermática y fragmentación del ADN en los genomas nucleares y mitocondriales. El test estrés espermático modificado (MOST) estima la resistencia espermática a la lipoperoxidación. El incremento de ERO en hombres con trastornos reproductivos y su correlación con alteraciones seminales revelan la importancia de la evaluación del EO en el estudio integral del hombre que consulta por infertilidad.
Data from Medline, Science Direct, Endocrine Society and organisms of information of Argentina Society of Andrology were used to study and interpret the effect of oxidative stress (OS) in the reproductive process. The living things use the oxygen for energy they release reactive oxygen species (ROS). There andrology situations enviromental factors, chemical agents that increase of proinflammatory cytokines, OS generated by altering the regulation of spermatogenesis process. The spermatozoids due to high concentration of polyunsaturated fatty acids (PFA) in its membrane are sensitive to the oxidative damage induced for ROS and it also contain low levels of antioxidant enzymes for their limited cytoplasm. OS induced peroxidative damage in sperm membrane and DNA fragmentation in the nuclear and mitochondrial genomes. The modified sperm stress test (MOST) estimates sperm resistance to lipoperoxidation. The ROS increased in men with reproductive disorders and its correlation with seminal alterations reveal the importance of the evaluation of OS in the study of man consulting for infertility.
RESUMO
Se relacionó la exposición ocupacional a agroquímicos con parámetros seminales vinculados con la espermatogénesis. En 104 muestras seminales de pacientes con infertilidad idiopática y 20 a 45 años de edad del Servicio de Reproducción Hospital Centenario (Rosario, Argentina), se efectuó espermograma y estudios funcionales según normas OMS (1999). Se agruparon en G1 (n=42) trabajadores expuestos a agroquímicos y G2 (n=62) hombres sin riesgo espermatogénico. La concentración espermática se determinó en cámara de Neubauer, con Papanicolaou se evaluó la concentración de células germinales y la morfología espermática. Se observó en concentración espermática (espermatozoides.10(6)/ml) G1: 21,1±7,1; G2: 41,6±9,2; morfología ( por ciento espermatozoides normales) G1: 5,2±1,3; G2: 8,2±3,6 y células germinales.10(6)/ml en G1: 0,91±0,51; G2: 0,32±0,21. La concentración espermática fue menor en G1, las alteraciones morfológicas y células germinales mayores en G1. Estos resultados indican que la exposición a agroquímicos altera la espermatogénesis y es un factor a considerar cuando se estudia infertilidad masculina.
The occupational exposition to agrochemicals was related with sperm parameters linked with spermatogenesis. In 104 semen samples of patients with idiopatic infertility of the Reproduction Service of Centenario Hospital (Rosario, Argentina), sperm study and functional tests according to WHO (1999) have been carried out. Two groups were formed: G1 (n=42) workers exposed to agrochemicals; G2 (n=62) men with no spermatogenic risk. The sperm concentration was determined in Neubauer camera, the concentration of germinal cells and the sperm morphology were evaluated with Papanicolaou. The results were: sperm concentration (spermatozoids 10(6)/ml) G1: 21.1 ± 7.1; G2: 41.6 ± 9.2; morphology ( percent normal spermatozoids) G1: 5.2 ± 1.3; G2: 8.2 ± 3.6; and germinal cells 10(6)/ml in G1: 0.91 ± 0.51; G2: 0.32 ± 0.21. The sperm concentration was lower in G1, major morphological changes and germinal cells in G1. This results show that the exposition to agrochemicals alters the spermatogenesis, it is a factor to consider in the male infertility.
RESUMO
La membrana espermática tiene ácidos grasos insaturados que la tornan vulnerable al ataque de sustancias oxígeno reactivas. Los espermatozoides poseen sistemas protectores, pero un desbalance entre pro y antioxidantes produce “estrés oxidativo". Objetivo: estudiar en semen de hombres infértiles, el efecto del estrés oxidativo sobre la membrana y núcleo espermático. Se analizaron muestras seminales de 142 hombres infértiles. Se efectuó espermograma, se seleccionaron 83 muestras sin aglutinación ni hiperviscosidad y con concentración espermática mayor a 5 x 10 6 /ml. Se estudió la membrana espermática con Test Hipoosmótico, la condensación cromatínica con Azul de Anilina y el ADN con Naranja de Acridina. Para estrés oxidativo se aplicó el Test MOST (movilidad traslativa final/movilidad traslativa inicial) que evalúa la pérdida de movilidad de los espermatozoides luego de ser incubados por 4 hs. en baño de agua a 40 °C. Se agruparon las muestras en G1: MOST mayor o igual a 0.40 (normal) y G2: MOST menor a 0.40 (anormal). El análisis estadístico demostró diferencia significativa (p<0.003) en las 3 pruebas funcionales. El estrés oxidativo altera estructuras espermáticas esenciales.
The spermatozoids have in its plasmatic membrane high concentrations of unsaturated fatty acids, vulnerable to the attack of the reactive oxygen substances. The male gamete has protective systems, the distortion between pro and antioxidizers produces “oxidative stress". Objective: to study samples of semen from infertile men, the effect of oxidative stress on the sperm membrane and the nucleus. 142 semen samples were analyzed from infertile men. The sperm study was evaluated according to OMS and 83 samples without agglutination and hiperviscosity with concentration of spermatozoids more of 5 x 10 6 /ml were chosen. The sperm membrane was studied with the Hipoosmotic Test, the maturity of chromatina with blue aniline and the nuclear AND with acridine orange. The EO was evaluated with the MOST test (motility end/motility initial) measuring the loss of motility of the spermatozoids incubated at 40ºC in a water bath for 4 hours, separating the samples in : G1 : MOST major or equal to 0.40 (normal) G2 : MOST under to 0.40 ( abnormal). The statistic analysis of the three tests showed significant difference (p<0.003). The oxidative stress affects essential structures.
RESUMO
OBJECTIVES: To investigate the presence of macrophages in human semen samples and the function they carry out in the seminal fluid. Their presence was studied in relation to spermatic morphology, percentage of spermatozoids with native DNA, and presence of antispermatic antibodies. METHODS AND RESULTS: The work was performed with semen samples from 31 unfertile males from 63 couples in which the "female factor" was ruled out as the cause of infertility. Sperm study according to WHO (1992) was carried out in all samples, in addition to: DNA study with acridine orange as fluorocrom, macrophage concentration by neutral red in a Neubauer camera, and detection of antispermatic antibodies with a mixed agglutination test (TAC II) (validated with Mar Screen-Fertility technologies). Sperm morphology was evaluated by Papanicolaou test. 19/31 selected sperm samples (61.3%) showed increased concentration of macrophages, 13 of them (41.9%) with denaturalized DNA, and 8 (25.8%) abnormal morphology. Six samples showed increased macrophage concentration and predominance of native DNA, whereas 11 samples showed increased macrophages and abnormal morphology. Among 18 (58.1%) samples showing antispermatic antibodies 14 (77.7%) had an increased concentration of macrophages. Statistical analysis resulted in a high correlation between macrophage concentration and increased percentage of spermatozoids with denaturalized DNA (p < 0.05). An increased concentration of macrophages is associated with the presence of antispermatic antibodies (p < 0.05). There was not evidence of significant association between concentration of macrophages and percentage of morphologically normal spermatozoids (p < 0.05). CONCLUSIONS: We can conclude that macrophages are present in human semen and participate in immunovigilance contributing to improve the seminal quality.