Postnatal Exposure to Exogenous progesterone Exacerbates the Prenatally-Induced Abnormal Sperms Production and Function in Rats / La exposición Posnatal a la Progesterona Exógena Exacerba la Producción y Función Anormales de Espermatozoides Inducidas Prenatalmente en Ratas

SUMMARY: This study aimed at clarifying the impact of long-term prenatal and postnatal exposure to exogenous progesterone on sperm production and function, relative sex organs weights, and the levels of the relevant hormones in rats. Sixty male Wistar rats were included and classified into three groups (n=20 in each). A test I group had mature rats born to dams treated with progesterone prenatally. A test II group included rats exposed to progesterone during prenatal as well as postnatal periods, and a control group had rats treated with a placebo (olive oil). The test groups revealed a significant reduction in sperm count, motility, and viability with higher abnormal forms than the control group (P< 0.05). Similarly, the test groups revealed significantly lower serum testosterone and higher FSH and LH levels (P< 0.001). Interestingly, the test II group showed pronounced sperm abnormalities, an alarming decrease in sperm viability and motility, and a significant accretion in the relative testicular weight compared to the test I group (p <0.001). Long-term (prenatal and early postnatal) treatment with synthetic progesterone hurts sperm quantity and quality, adversely affecting future male fertility.

Este estudio tuvo como objetivo aclarar el impacto de la exposición prenatal y posnatal a largo plazo a la progesterona exógena en la producción y función de los espermatozoides, el peso relativo de los órganos sexuales y los niveles de las hormonas relevantes en ratas. Sesenta ratas macho Wistar fueron incluidas y clasificadas en tres grupos (n=20 en cada uno). Un grupo de prueba I tenía ratas maduras nacidas de madres tratadas con progesterona prenatalmente. Un grupo de prueba II incluyó ratas expuestas a progesterona durante los períodos prenatal y posnatal, y un grupo de control tenía ratas tratadas con un placebo (aceite de oliva). Los grupos de prueba revelaron una reducción significativa en el recuento, la motilidad y la viabilidad de los espermatozoides con formas anormales más altas que el grupo de control (P < 0,05). De manera similar, los grupos de prueba revelaron niveles significativamente más bajos de testosterona sérica y niveles más altos de FSH y LH (P < 0.001). Curiosamente, el grupo de prueba II mostró anormalidades espermáticas pronunciadas, una disminución alarmante en la viabilidad y motilidad de los espermatozoides y una acumulación significativa en el peso testicular relativo en comparación con el grupo de prueba I (p <0.001). El tratamiento a largo plazo (prenatal y posnatal temprano) con progesterona sintética daña la cantidad y la calidad del esperma, lo que afecta negativamente la futura fertilidad masculina.

Cyperus esculentus L. protects testis and sperm morphology of hyperglycaemic rats / Cyperus esculentus L. protege la morfología de los testículos y los espermatozoides de ratas hiperglicémicas

Cyperus esculentus L. (tiger nut) is a tuberous plant that promotes and protects reproductive functions, which are usually hampered in diabetics. The present study investigated the effect of Cyperus esculentus tuber extract (CETE) on testicular histology and sperm viability of alloxan-induced hyperglycaemic Wistar rats. Twenty-five adult male Wistar rats weighing 150-200g and grouped into five (n=5): Group 1, the control, administered tap water (20mL/kg), while groups 2-5 were administered a single intraperitoneal dose (120mg/kg b.w.) of alloxan, and each further received orally tap water (20mL/kg), CETE (100mg/kg), CETE (500 mg/kg) and metformin (500 mg/kg), respectively for 21 days. The animals were sacrificed, their sperm collected for analysis, while the testes were harvested, and processed for histology. Results showed significantly increased (p<0.05) blood glucose and testosterone, and significantly decreased (p<0.05) sperm pH, motility, count, morphology and density, as well as disruptions and hypertrophy of the spermatogenic and Sertoli cells of the hyperglycaemic group. There were significant (p<0.05) blood glucose decline, while the sperm parameters and testicular weight improved with normal testicular histology in the 100 mg/kg CETE, 500 mg/kg CETE, and metformin-treated groups compared to the control and hyperglycaemic group. Treatment with CETE showed blood glucose amelioration and improved sperm quality, as well as testicular damage attenuation.

Cyperus esculentus L. es una planta tuberosa que promueve y protege las funciones reproductivas, que generalmente se ven afectadas en los diabéticos. El presente estudio investigó el efecto del extracto de tubérculo de Cyperus esculentus (CETE) sobre la histología testicular y la viabilidad de los espermatozoides de ratas wistar con hiperglicemia inducida por alloxan. Veinticinco ratas Wistar macho adultas que pesaban 150-200 g y se agruparon en cinco (n = 5): el grupo 1, el control, administró agua del grifo (20ml / kg), mientras que los grupos 2-5 se les administró una dosis intraperitoneal única (120 mg / kg p.v.) de alloxan, y agua del grifo por vía oral (20ml/kg), CETE (100 mg/kg), CETE (500 mg/kg) y metformina (500 mg/kg), respectivamente durante 21 días. Los animales fueron sacrificados, su esperma recolectada para su análisis, mientras que los testículos fueron retirados y procesados para histología. Los resultados mostraron un aumento significativo (p<0,05) de la glucosa en sangre y la testosterona, y una disminución significativa (p<0,05) del pH, la motilidad, el recuento, la morfología y la densidad de los espermatozoides, así como interrupciones e hipertrofia de las células espermatogénicas y sertoli del grupo hiperglucémico. Hubo una disminución significativa (p<0,05) de la glucosa en sangre, mientras que los parámetros espermáticos y el peso testicular mejoraron con la histología testicular normal en los grupos de 100 mg / kg de CETE, 500 mg / kg de CETE y tratados con metformina en comparación con el grupo de control e hiperglucémico. El tratamiento con CETE mostró una mejora de la glucosa en sangre y una mejora de la calidad de los espermatozoides, así como atenuación del daño testicular.

Buchholzia coriacea (wonderful kola) seeds induce male reproductive toxicity by suppressing the pituitary-gonadal axis in Wistar rats

The methanolic extract of Buchholzia coriacea seeds (MEBC) has been reported to induce male reproductive toxicity by decreasing sperm parameters and fertility index. To elucidate the possible mechanism(s), the effects of graded doses of MEBC on sex hormones and sperm profile were investigated in this study. The MEBC (e.g., 50, 200, 400, and 600 mg/kg) was administered daily (p.o.) to male Wistar rats for 6 weeks, while a concurrent control group received distilled water (vehicle). Then, the animals were sacrificed under sodium pentobarbital anaesthesia. Weights of organs were recorded, and the sperm profile was determined microscopically. Testosterone, luteinizing hormone (LH), and follicle stimulating hormone (FSH) were assayed from the obtained serum using the ELISA technique. Sperm motility was significantly reduced by MEBC (i.e., 50 and 200 mg/kg), and sperm count reduced in all treated groups in a dose-dependent manner compared with that of the control. Serum testosterone, LH, and FSH decreased in treated rats. A histopathological examination of testes showed a considerable depletion and necrosis of the epithelium of seminiferous tubules. The result suggests that Buchholzia coriacea seeds induce male reproductive toxicity by suppressing the pituitary-gonadal axis.

Efecto de la borra de café sobre la biológica espermática: aproximación in vitro / Effect of spent coffee grounds on sperm biology: in vitro approach

OBJETIVO: El objetivo de este trabajo fue establecer el efecto de la borra de café sobre la movilidad y los parámetros funcionales de los espermatozoides humanos in vitro. MATERIALES Y MÉTODOS: La borra de café, un subproducto obtenido en establecimientos especializados en la preparación de café soluble a base de grano, se diluyo en tampón fosfato salino y se mezcló en proporciones iguales con las muestras de semen de 16 voluntarios aparentemente sanos. A cada muestra se le determinó el efecto sobre la movilidad espermática en función del tiempo (30, 60, 90 y 120 minutos, n=16) y sobre los parámetros funcionales (n=6) por medio de citometría de flujo: potencial de membrana mitocondrial, producción de especies reactivas de oxígeno y lipoperoxidación de la membrana espermática. RESULTADOS: La incubación de los espermatozoides con la borra de café evidencio un cambio positivo en la movilidad espermática. Adicionalmente, la incubación con la borra de café incremento significativamente el potencial de membrana mitocondrial en los espermatozoides. CONCLUSIÓN: La borra de café, seguramente debido a los compuestos antioxidantes, afecta positivamente la movilidad espermática aumentando el potencial de membrana mitocondrial. Por lo tanto, esto es un paso inicial en la búsqueda de un suplemento de origen natural que aumente la calidad seminal.

OBJECTIVE: The objective of this work is to establish the effect of spent coffee grounds on the motility and functional parameters of human spermatozoa, in vitro. MATERIALS AND METHODS: Spent coffee grounds, a by-product obtained in specialized establishments in the preparation of soluble coffee based on grain, was diluted in saline phosphate buffer and mixed in equal proportions with semen samples from 16 apparently healthy volunteers. Each sample was determined the effect on sperm motility as a function of time (30, 60, 90 and 120 minutes, n=16) and on functional parameters (n=6) by means of flow cytometry: mitochondrial membrane potential, reactive oxygen species production and membrane lipoperoxidation. RESULTS: The incubation of the spermatozoa with the spent coffee grounds showed a positive change in sperm motility. Additionally, incubation with spent coffee grounds significantly increased the mitochondrial membrane potential in human sperm cells. CONCLUSION: Spent coffee grounds, probably due to antioxidant compounds, positively affects sperm motility by increasing mitochondrial membrane potential. Therefore, this is an initial step in the search for a supplement of natural origin that increases seminal quality.

Post-finasteride syndrome

Abstract Finasteride is a 5α-reductase enzyme inhibitor that has been approved for the treatment of male androgenic alopecia since 1997. Over time, it has been considered a safe and well-tolerated drug with rare and reversible side effects. Recently there have been reports of adverse drug-related reactions that persisted for at least three months after discontinuation of this drug, and the term post-finasteride syndrome arose. It includes persistent sexual, neuropsychiatric, and physical symptoms. Studies to date cannot refute or confirm this syndrome as a nosological entity. If it actually exists, it seems to occur in susceptible people, even if exposed to small doses and for short periods, and symptoms may persist for long periods. Based on currently available data, the use of 5α-reductase inhibitors in patients with a history of depression, sexual dysfunction, or infertility should be carefully and individually assessed.

Addition of synthetic amino acids in sheep semen improves its quality after cryopreservation / Adição de aminoácidos sintéticos no sêmen ovino melhora a qualidade espermática após a criopreservação do sêmen

Some amino acids can protect mammalian sperm cells against oxidation during thermal stress caused by freezing/thawing. Thus, the objective was to evaluate the protective action of the association of the amino acids L-proline (Pro) and L-glutamine (Glu) against the cryoinjury caused to sheep sperm after cryopreservation. Eight ejaculates were collected from four sheep (n=32) and diluted in Tris-Egg Yolk-Glycerol until the final concentration of 200 x106 sptz/mL and kept in a water bath at 32 °C. The amino acids were added as follows: control (without adding amino acids), Pro+Glu 1 (100 µM Pro + 500 µM Glu), Pro+Glu 2 (300 µMPro + 1000 µM Glu), Pro+Glu 3 (500 µM Pro + 1500 µM Glu) and Pro+Glu 4 (700 µM Pro + 2000 µM Glu). Afterwards, the semen was cooled to 5 °C for 2 h, after that period, filled in 0.5 mL straws and then placed under liquid nitrogen vapor (N2L), 8 cm from the liquid sheet for 15min, and then immersed on the N2L. The samples were analyzed for sperm motility, plasma membrane and acrosomal membrane integrity, mitochondrial activity and binding test. The variables were subjected to the normality tests (Lilliefors test) and homoscedasticity tests (Cochran and Bartlett test), afterwards the variables of normal distribution were subjected to analysis of variance and the means compared by the Tukey test with a significance level of 5%. The Pro+Glu 3 group exhibited sperm with a greater (P<0.05) motility after thawing. In addition, the highest percentage of plasma and acrosomal membrane integrity were obtained using Pro+Glu 1, Pro+Glu 2 and Pro+Glu 3; and Pro+Glu 2 and Pro+Glu 3, respectively. Amino acids also kept mitochondrial activity high compared to the control, with Pro+Glu 3 resulting in greater activity (P<0.05). Sperm viability was higher (P<0.05) with the use of Pro+Glu 2 and Pro+Glu 3 than in the control. The number of sperm that showed the ability to bind to the egg yolk perivitelline membrane was higher (P<0.05) in semen treated with amino acids. It is concluded that the addition of synthetic amino acids in the semen of sheep before cryopreservation improves sperm quality and fertilization potential and can thus be added in cryopreservation protocols.

Alguns aminoácidos podem proteger as células espermáticas de mamíferos contra a oxidação durante o estresse térmico causado na congelação/descongelação. Dessa forma, objetivou-se avaliar a ação protetora da associação dos aminoácidos L-prolina (Pro) e L-glutamina (Glu) contra as crioinjúrias causadas aos espermatozoides de ovino após a criopreservação. Foram coletados oito ejaculados de quatro carneiros (n=32) e diluídos em Tris-Gema de ovo-Glicerol até a concentração final de 200 x106 sptz/mL e, mantidos em banho maria a 32 °C. Os aminoácidos foram adicionados da seguinte forma: controle (sem adição de aminoácidos), Pro+Glu 1 (100 µM Pro + 500 µM Glu), Pro+Glu 2 (300 µM Pro + 1000 µM Glu), Pro+Glu 3 (500 µM Pro + 1500 µM Glu) e Pro+Glu 4 (700 µM Pro + 2000 µM Glu). Depois, o sêmen foi resfriado a 5 °C por 2 h, após esse período, envasado em palhetas de 0,5 mL e então acondicionado sob vapor de nitrogênio líquido (N2L), a 8 cm da lâmina líquida por 15 min, e depois imersos no N2L. As amostras foram analisadas quanto à motilidade espermática, integridade da membrana plasmática e da membrana acrossomal, atividade mitocondrial e teste de ligação. As variáveis foram submetidas aos testes de normalidade (Teste de Lilliefors) e homocedacidade (Teste de Cochran e Bartlett), posteriormente as variáveis de distribuição normal foram submetidas à análise de variância e as médias comparadas pelo teste de Tukey com nível de significância de 5%. O grupo Pro+Glu 3 exibiu espermatozoides com uma maior (P<0,05) motilidade após o descongelamento. Além disso o maior percentual de integridade da membrana plasmatica e acrossomal foram obtidos utilizando Pro+Glu 1, Pro+Glu 2 e Pro+Glu 3; e Pro+Glu 2 e Pro+Glu 3, respectivamente. Os aminoácidos também mantiveram alta a atividade mitocondrial em comparação com o controle, com Pro+Glu 3 resultando numa maior atividade (P<0,05). A viabilidade dos espermatozoides foi maior (P<0,05) com o uso de Pro+Glu 2 e Pro+Glu 3 do que no controle. O número de espermatozoides que apresentaram à capacidade de ligação a membrana perivitelina da gema de ovo foi maior (P<0,05) no sêmen tratado com aminoácidos. Conclui-se que, a adição dos aminoácidos sintéticos no sêmen de ovinos antes da criopreservação melhora a qualidade espermática e o potencial fecundante, podendo assim serem adicionados em protocolos de criopreservação.

Efecto del zumo de sandía (citrullus lanatus) en el estrés oxidativo en espermatozoides humanos / Effect of watermelon juice (citrullus lanatus) on oxidative stress in human sperm

INTRODUCCIÓN: Las alteraciones reproductivas de causa masculina relacionadas con el estrés oxidativo son cada día más estudiadas y dan cuenta de causas de infertilidad diagnosticada como idiopáticas. El objetivo del presente trabajo fue evaluar el efecto del zumo de sandía sobre los parámetros seminales convencionales y funcionales in vitro e in vivo. MATERIALES Y MÉTODOS: Cinco muestras de espermatozoides puros fueron incubados con peróxido de hidrógeno (H2O2, 5mM) y 0,45% de extracto de sandía, se determinó la movilidad espermática al tiempo 0, 30 y 60 minutos. En los ensayos in vivo se incluyeron 20 individuos a los cuales se les determinaron los parámetros espermáticos convencionales, funcionales y la capacidad antioxidante del plasma seminal por microscopía, citometría y espectrofotometría en los días 0, 7 y 15 después de iniciar el consumo diario de 16 onzas de zumo de sandía. RESULTADOS: El extracto de sandía protege a los espermatozoides del efecto deletéreo del H2O2 sobre la movilidad espermática. Además, el consumo regular de jugo de sandía disminuye la lipoperoxidación de la membrana espermática, la producción intracelular de especies reactivas del oxígeno, el índice de fragmentación del ADN el día 15 y la capacidad antioxidante el día 7 y 15. CONCLUSIONES: El extracto de sandía genera un efecto protector sobre los espermatozoides humanos in vitro, protegiendo su movilidad del efecto negativo del H2O2. Además, si bien el consumo regular de zumo de sandía no mejora los parámetros seminales convencionales, si mejora algunos parámetros funcionales relacionados con el estrés oxidativo.

OBJETIVE: Male reproductive alterations related to oxidative stress are increasingly studied and account for causes of infertility diagnosed as idiopathic. The aim of this work was to evaluate the effect of watermelon juice on conventional and functional seminal parameters in vitro and in vivo. MATERIALS AND METHODS: Five samples of pure sperm were incubated with hydrogen peroxide (H2O2, 5mM) and 0.45% watermelon extract, sperm motility was determined at time 0, 30 and 60 minutes. In vivo assays, 20 individuals were included. Conventional and functional sperm parameters, and antioxidant capacity of seminal plasma using microscopy, cytometry and spectrophotometry were determined on days 0, 7 and 15 after starting daily consumption of 16 ounces of watermelon juice. RESULTS: Watermelon extract protects sperm cells from the deleterious effect of H2O2 on sperm motility. In addition, regular consumption of watermelon juice decreases sperm membrane lipoperoxidation, intracellular production of reactive oxygen species, DNA fragmentation index on day 15 and antioxidant capacity on day 7 and 15. CONCLUSION: Watermelon extract generates a protective effect on human sperm in vitro, protecting sperm motility from the negative effect of H2O2. In addition, although regular consumption of watermelon juice does not improve conventional seminal parameters, it does improve some functional parameters related to oxidative stress.

Improvement of fertility parameters with tribulus terrestris and anacyclus pyrethrum treatment in male rats

ABSTRACT Objective Anacyclus Pyrethrum (AP) and Tribulus Terrestris (TT) have been reported as male infertility treatment in several studies; however, in Iranian traditional medicine these two plants are prescribed simultaneously. In this study, we aimed to determine the effects of AP and TT extracts both separately and simultaneously on the male Wistar rat fertility parameters. Materials and Methods 32 male Wistar rats were divided into 4 groups: Control, TT, AP, and AT treated groups. Treatment continued for 25 days and rats were weighed daily. Their testes were dissected for histological studies. Sperm analysis including sperm count, viability and motility were performed. Serum was obtained to evaluate testosterone, LH and FSH levels. Histological studies were conducted to study Leydig, and Sertoli cells, spermatogonia and spermatid cell numbers, and to measure seminiferous diameter and epithelium thickness. Results Sperm count increased in all the treatment groups. Sperm viability and motility in AT and AP groups were elevated. TT and AT groups showed significantly increased testosterone level compared to control group (P=004, P=0.000, respectively) and TT, AP and AT treatment groups showed increased LH level (P=0.002, P=0.03 and P=0.000, respectively) compared to control, while only AT group showed increased FSH (p=0.006) compared to control. Histological studies showed significant increase of spermatogonia, Leydig and Sertoli cell numbers and epithelial thickness in AT group compared to other groups. All the treatment groups had higher number of Leydig, spermatogonia and spermatid cells. Conclusion TT and AP improved sexual parameters; however, their simultaneous administration had higher improving effects on studied parameters.

Antioxidant action of alpha lipoic acid on the testis and epididymis of diabetic rats: morphological, sperm and immunohistochemical evaluation

ABSTRACT Introduction Chronic hyperglycemia is caused by diabetes mellitus-committed genital morphophysiology, and oxidative stress is one of the main factors involved in this process. Alpha lipoic acid (ALA) can prevent metabolic and morphological changes in diabetic individuals. Objectives In present study, we evaluated the effects of regular ALA consumption on the spermatogenesis and histoarchitecture in the male genital system of diabetic rats. Materials and Methods Thirty-two Wistar rats were divided into groups: Control (CG); Diabetic Control (DCG), receiving commercial diet: ALA Group (ALAG) and Diabetic ALA Group (DALAG), fed diets with added ALA (300 mg/Kg bw). The diabetic groups received a single injection of streptozotocin (60 mg/kg). After sixty days of the diet, the animals were euthanized, and semen, testis and epididymis samples were collected. A histomorphometric analysis was performed to determine the epithelial height, tubular and luminal diameter, tubular and luminal area of seminiferous tubules and each epididymal region. Sertoli cells were evidenced using the antivimenti antibody and were quantified. The results were statistically analyzed by the ANOVA test. Results At the end of the experiment, the DALAG glycemia was significantly lower than DCG. The histomorphometric parameters of the seminiferous and epididymal tubules did not show improvement in the DALAG. However, there was an improvement in the DALAG in terms of the concentration, motility and percentage of spermatic pathologies, as well as in the number of Sertoli cells (p<0.001). Conclusions The results demonstrated that supplementation with the ALA antioxidant retards testicular lesions and preserve the process of spermatogenesis in diabetes.

Protective Effect of Rosa Canina Extract Against Doxorubicin-induced Testicular Toxicity in Mice

Abstract The present study was aimed to investigate the in vivo effects of Rosa canina extract on doxorubicin-induced testicular toxicity in mice for the first time. Male NMRI mice were randomly divided into six treatment groups (10=per group) as follows: (I) vehicles, (II) doxorubicin alone (3 mg/kg, i.p. on days 7, 14 and 21), (III and IV): Rosa canina extract alone (100 mg/kg and 200 mg/kg per day, i.p. for 28 days), (V and VI) Rosa canina extract plus doxorubicin (each dose given 1 h post Rosa canina). Doxorubicin-treated mice displayed smaller body and testicular weights, decreased serum levels of testosterone, loss in the number of germ cells and Sertoli cells, and reduced sperm count, viability, morphology and motility. Doxorubicin treatment significantly decreased the mean testis diameter, seminiferous tubular diameter and seminiferous epithelial height and increased seminiferous luminal diameter. However, Rosa canina pretreatment could effectively improve all of these abnormalities in doxorubicin- treated mice. The treatment with a higher dose of the extract (200 mg/kg) was more effective compared to doxorubicin and the lower dose of the extract. These findings suggested that the Rosa canina extract has protective effects against doxorubicin-induced reproductive toxicity.

Effects of Leucaena leucocephala (lamk. ) shoot tips plus young leaf extract containing mimosine on reproductive system of male rats / Efectos de los brotes de Leucaena leucocephala (lamk. ) y del extracto de hojas jóvenes que contiene mimosina en el sistema reproductivo de ratas macho

This study aimed to determine the mimosine level and examine the male reproductive toxicity effects of Leucaena leucocephala (LL) shoot tips plus young leaf extract. Mimosine level in LL extract was determined by thin layer chromatography before administration in animals. Male rats were divided into control and LL (1,500 mg/KgBW) groups (n = 6). After 60 days of experiment, serum sex hormones, sperm quality, and testicular histopathology were assayed and observed. Malondialdehyde (MDA) level and expressions of steroidogenic acute regulatory (StAR) and phosphorylated proteins in testicular lysate were examined by western blotting. Results showed that mimosine levels in LL extract was 17.35 ± 1.12 % of dry weight. LL significantly decreased FSH & LH levels, sperm qualities, and seminiferous tubule diameter compared to the control (p<0.05). Seminiferous tubular atrophies, germ cell sloughing, and degenerations were observed in LL group. In addition, testicular MDA level and StAR protein expression were significantly decreased in LL group. LL extract could increase the expression of a 50 kDa phohorylated protein in testicular lysate. In conclusion, LL extract has mimosine and reproductive toxicity effects on males.

Este trabajo tuvo como objetivo determinar el nivel de mimosina y examinar los efectos de la toxicidad reproductiva de los brotes de Leucaena leucocephala (LL), más el extracto de hojas jóvenes, en ratas macho. El nivel de mimosina en el extracto de LL se determinó mediante cromatografía en capa fina antes de la administración en animales. Las ratas se dividieron en grupos de control y LL (1,500 mg / kgBW) (n = 6). Después de 60 días, se analizaron y observaron las hormonas sexuales séricas, la calidad de los espermatozoides y la histopatología testicular. A través de Western Blot se examinaron el nivel de malondialdehído (MDA), las expresiones de reguladores agudos esteroidogénicos (StAR) y las proteínas fosforiladas en el lisado testicular. Los resultados mostraron que los niveles de mimosina en el extracto de LL fueron 17.35 ± 1.12 % del peso seco. LL disminuyó significativamente los niveles de FSH y LH, la calidad de los espermatozoides y el diámetro de los túbulos seminíferos en comparación con el control (p <0,05). Se observaron atrofias en los túbulos seminíferos, desprendimiento de células germinales y degeneraciones en el grupo LL. Además, el nivel de MDA testicular y la expresión de la proteína StAR se redujeron significativamente en el grupo LL. El extracto de LL podría aumentar la expresión de la proteína fosforilada de 50 kDa en el lisado testicular. En conclusión, el extracto de LL tiene mimosina y efectos de toxicidad reproductiva en los hombres.

Antioxidant capacity of Momordica charantia extract and its protective effect on testicular damage in valproic acid-induced rats / Capacidad antioxidante del extracto de Momordica charantia y su efecto protector sobre el daño testicular en ratas inducidas con ácido valproico

Although Momordica charantia (MC) has preventive effects on tissue injuries, antioxidant capacity and protective effect of MC pulp and peel (MCP) on valproic acid (VPA)-testicular damage have never been reported. Fresh MCPs were aqueous extracted and determined for antioxidant capacity and momordicine I level by HPLC. Male rats were divided into 5 groups (control, VPA (500 mg/kgBW), MCP20/40/ or 80 mg/kgBW+VPA). In 30 experimental days, animals were pretreated with different doses of MCPs for 20 days before VPA injection for 10 consecutive days. Sperm concentration, testosterone hormone, and testicular histology of all groups were investigated. Expressions of testicular tyrosine phosphorylated and steroidogenic acute regulatory (StAR) proteins were examined by Western blot. Results showed that MCP contains TPC (39.24±0.65 ug/mg garlic acid), antioxidant capacities (FRAP=33.08±0.21 ug/ mg ascorbic acid equivalent, IC50 of DPPH=389.8±3.20 ug/ml), and momordicine I (404.9 mg/g MCP). Sperm concentration in MCP80+VPA group was increased as compared to VPA group. Testosterone level in MCP treated groups was significantly increased. MCP protected testicular damage and could prevent the decrease of StAR and a 50-kDa phosphorylated protein expression in VPAtreated testis. In conclusion, MCP has antioxidant activities and can prevent male reproductive toxicity in VPA-induced rats.

A pesar que la Momordica charantia (MC) tiene efectos preventivos sobre las lesiones en los tejidos, capacidad antioxidante y un efecto protector de la pulpa y la cáscara de MC (CMC) sobre el ácido valproico (AVP), aún no se ha informado efectos sobre el daño testicular. Las CMC frescas fueron extraídas de forma acuosa y se determinó la capacidad antioxidante y el nivel de Momordicina I por HPLC. Las ratas machos se dividieron en 5 grupos: control, AVP (500 mg/kg de peso corporal), CMC20 / 40 / u 80 mg/kg de peso corporal + AVP . En 30 días experimentales, los animales fueron pretratados con diferentes dosis de CMC durante 20 días antes de la inyección de AVP durante 10 días consecutivos. Se investigó la concentración de espermatozoides, la hormona testosterona y la histología testicular de todos los grupos. Las expresiones de proteínas reguladoras agudas (StAR) fosforiladas con tirosina y esteroidogénicas testiculares se examinaron mediante inmunotransferencia de tipo Western. Los resultados mostraron que CMC contiene TPC (39.24 ± 0.65 ug / mg de ácido de ajo), capacidades antioxidantes (FRAP = 33.08 ± 0.21 ug / mg de ácido ascórbico equivalente, IC50 de DPPH = 389.8 ± 3.20 ug / ml) y momordicina I (404.9 mg) / g CMC). La concentración de esperma en el grupo MCP80 + AVP aumentó en comparación con el grupo AVP. El nivel de testosterona en los grupos tratados con CMC aumentó significativamente. La CMC protegió el daño testicular y pudo prevenir la disminución de StAR y una expresión de proteína fosforilada de 50 kDa en los testículos tratados con AVP. En conclusión, la CMC tiene efectos antioxidantes y puede prevenir la toxicidad reproductiva en ratas machos inducidas por VPA.

Therapeutic effect of thymoquinone against methotrexate- induced damage on sperm parameters in mice / Efecto terapéutico de la tiroyoquinona contra el daño inducido por metotrexato en los parámetros espermáticos en ratones

Methotrexate drug is commonly used to treat cancer; it is known to cause reproductive damage. Thymoquinone, as a natural component of herbs has many healthy benefits shown in researches. The present study aimed to investigate probable therapeutic effects of Thymoquinone against Methotrexate-induced damage on sperm parameters in mice. In this experimental study, 30 male mice (25-30 g) were divided into five groups of six in each group. The mice were received normal saline (control group), Methotrexate (20 mg/kg), Methotrexate (20 mg/kg) + Thymoquinone (2, 10 and 20 mg/kg) by intraperitoneal injection. On the day after the last injection, the sperm parameters including motility, viability and count of sperms were assessed. Data analysis was performed using one-way ANOVA followed by Tukey test. Methotrexate alone showed a significant reduction in sperm parameters compared to the control group (P=0.00). In groups treated with Methotrexate and Thymoquinone, sperm parameters (motility ,viability, count sperm) did not show any significant differences with control group (P=0.00). Thymoquinone, as a potent antioxidant, could compensate for the toxicity induced by Methotrexate. These medical trends may be useful for diminishing the side effects of Methotrexate on the male reproductive system.

El metotrexato es un fármaco utilizado comúnmente para tratar el cáncer pero además causa daño en los órganos reproductivos. Durante las investigaciones se ha demostrado que la timoquinona, un componente natural de las hierbas, tiene numerosos beneficios. El objetivo del estudio fue investigar el probable efecto terapéutico de la timoquinona contra el daño inducido por metotrexato, en los parámetros espermáticos en ratones. En este estudio experimental, se dividieron 30 ratones machos (25-30 g) en cinco grupos de seis en cada uno. Los ratones recibieron solución salina normal (grupo control), metotrexato (20 mg / kg), metotrexato (20 mg / kg) + timoquinona (2, 10 y 20 mg / kg) por inyección intraperitoneal. El día después de la última inyección, se evaluaron los parámetros espermáticos, incluida la motilidad, la viabilidad y el recuento de espermatozoides. El análisis de los datos se realizó utilizando test de ANOVA seguido de la prueba de Tukey. Durante el uso exclusivo de metotrexato se observó una reducción significativa en los parámetros espermáticos en comparación con el grupo control (P = 0.00). En los grupos tratados con metotrexato y timoquinona, los parámetros espermáticos (motilidad, viabilidad, conteo de espermatozoides) no mostraron diferencias significativas con el grupo control (P = 0.00). Como potente antioxidante, la timoquinona podría compensar la toxicidad inducida por metotrexato. Estas tendencias médicas pueden ser útiles para disminuir los efectos secundarios de metotrexato en el sistema reproductivo masculino.

Methotrexate changes the testicular tyrosine phosphorylated protein expression and seminal vesicle epithelia of adult rats / El metotrexato cambia la expresión de la proteína testicular-tirosina fosforilada y el epitelio vesicular seminal en ratas adultas

Methotrexate (MTX) is commonly used as a chemotherapy agent and immune system suppressant but its adverse effect on male reproductive system is still limited. This study aimed to investigate the effect of MTX on structure and functional proteins of testis and seminal vesicle. Adult male rats were divided into control and MTX groups (n =12). In 30 experimental days, the treated animals were injected with MTX (tail i.v., 75 mg/KgBW) at days 8 and 15. Then, the reproductive parameters and histology of both groups were examined. Thickness of seminal seminal vesicle epithelia was analyzed. Also, the expressions of testicular tyrosine phosphorylated proteins and steroidogenic acute regulatory (StAR) protein were investigated. The results showed that MTX could significantly decrease epididymal sperm concentration. In addition, the germ cell degeneration, increased spaces of interstitial tissues, and low epididymal sperm mass density were observed in MTX group. The thickness of seminal vesicle epithelia in MTX group was significantly lower than that of control group. Moreover, the intensity of testicular phosphorylated proteins of 31, 32, 72, and 85 kDas was significantly increased while of 42 and 47 kDas in MTX group was decreased as compared to control. The expression of testicular StAR protein in MTX group was also significantly decreased as compared to the control. In conclusion, MTX affects testicular and seminal tissues and changes testicular functional proteins in adult rats.

El metotrexato (MTX) se usa comúnmente como agente de quimioterapia y supresor del sistema inmunitario, pero su efecto adverso en el sistema reproductor masculino sigue siendo limitado. Este estudio tuvo como objetivo investigar el efecto del MTX sobre la estructura y las proteínas funcionales del testículo y la vesícula seminal. Ratas macho adultas se dividieron en grupos control y grupo con MTX (n = 12). En 30 días experimentales, a los animales tratados se les inyectó MTX (cola i.v., 75 mg / KgBW) los días 8 y 15. Luego, se examinaron los parámetros reproductivos y la histología de ambos grupos. Se analizó el espesor del epitelio de la vesícula seminal. Además, se investigaron las expresiones de la proteína tirosina testicular fosforilada y de la proteína reguladora aguda esteroidogénica (StAR). Los resultados mostraron que el MTX podría disminuir significativamente la concentración de espermatozoides epididimarios. Además, se observó la degeneración de las células germinales, el aumento de los espacios de los tejidos intersticiales y la baja densidad de masa del espermatozoide epididimal en el grupo de MTX. El grosor del epitelio de la vesícula seminal en el grupo MTX fue significativamente menor que el del grupo control. Además, la intensidad de las proteínas testiculares fosforiladas de 31, 32, 72 y 85 kDas aumentó significativamente, mientras que la de 42 y 47 kDas en el grupo MTX disminuyó en comparación con el control. La expresión de la proteína StAR testicular en el grupo MTX también se redujo significativamente en comparación con el control. En conclusión, el MTX afecta los tejidos testiculares y seminales y cambia las proteínas funcionales testiculares en ratas adultas.

Effects of folic acid administration on testicular ischemia/reperfusion injury in rats

Abstract Purpose: To determine the effect of folic acid (FA) on experimental testicular ischemia/reperfusion (I/R) in rats. Methods: Sixty male Wistar rats were randomly divided into 6 groups. The control group received physiologic saline orally. The sham-operated group received physiologic saline orally then exposed to midline laparotomy without clamping the IR. The I/R rats received oral gavage of the saline then subjected to 1h ischemia /24h reperfusion, period. In folic acid (2mg/kg+IR) rats received oral gavage of the FA (2mg/kg) then subjected to 1h I/24h R. groups 5-6 received FA (5 and 10 mg/kg), then subjected to 1 h I/24 h, respectively. At the end of the study, semen samples were collected for spermatozoa characteristics. The left testis was removed for histological analysis and superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione peroxidase (GPx) measurement. Results: Spermatozoa mobility, mortality (%) significantly decreased in I/R group (P<0.05). Dose dependent increase observed on spermatozoa mobility, mortality (%) using different levels of the FA (2, 5 and 10 mg/kg) treated rat (P<0.05). Tissue MDA levels significantly increased in I/R rat (P<0.05) while FA (2, 5 and 10mg/kg) in a dose dependent manner decreased I/R-induced MDA (P<0.05). Experimental I/R significantly decreased SOD and GPx activity (P<0.05). Administration of the FA (2, 5 and 10mg/kg) significantly increased tissue SOD and GPx activity in I/R rat (P<0.05). Seminiferous tubules degenerated and loss of spermatogenesis with few spermatocytes was observed in degenerated testis tubules in I/R rat. Orally administration of the FA (5 and 10 mg/kg) improved testis characteristics with few normal seminiferous tubules and spermatocyte in seminiferous tubules in experimental I/R-induced rat. Conclusion: The treatment of folic acid had a benefit effect against ischemia-reperfusion.

Organophosphorous pesticides: their effects on biosentinel species and humans: control and application in Chile / Pesticidas organofosforados: sus efectos en especies biosentinelas y humanos: control y aplicación en Chile

The environment is negatively affected by the increasing accumulation of both natural and man-made waste and by-products. Organophosphorous pesticides -malathion, diazinon and methamidophos- are used worldwide in pest control. The aim of this report is to review the effects of organophosphates on the male reproductive tract of mice, rats and earthworms, and to evaluate their projection into the human population. Assessing failures in the male reproductive system is an excellent in vivo biomarker to determine the level of toxicity of suspected pollutants. In rodents organophosphates cause decreased testicular weight and sperm density, abnormal tubular plugging and increased teratozoospermia. In earthworms they cause a significant decrease in body weight and alter the spermatheca, with an initial significant increase in immature sperm followed by a significant decrease in sperm count with high frequency of metachromasia. Given the environmental impact of these pesticides -and their potential effects on human health-, international and regional organizations are warning about the correct handling and managing of these substances during work-related and domestic exposures and about their relation to water sources and food, placing a greater emphasis on the school children population due to its higher vulnerability, reduced detoxification capacity, and their endocrine and physiological effects.

El medio ambiente se ve afectado negativamente por la creciente acumulación de desechos y subproductos naturales y artificiales. Los plaguicidas organofosforados malatión, diazinón y metamidofos, son usados en todo el mundo en el control de plagas. El objetivo de este trabajo fue revisar los efectos de los organofosforados en el tracto reproductivo masculino de ratones, ratas y lombrices de tierra, y evaluar su proyección en la población humana. La evaluación de fallas en el sistema reproductor masculino es un excelente biomarcador in vivo para determinar el nivel de toxicidad de los contaminantes químicos. En roedores, los organofosforados causan disminución del peso testicular y de la densidad espermática, obstrucción tubular anormal y aumento de la teratozoospermia. En lombrices de tierra causan una disminución significativa en el peso corporal y alteran la espermateca, con un aumento inicial significativo en espermatozoides inmaduros, seguido de una disminución significativa en el recuento de espermatozoides con alta frecuencia de metacromasia. Dado el impacto medioambiental de estos plaguicidas y sus efectos potenciales en la salud humana, las organizaciones internacionales y regionales advierten sobre el manejo y uso correctos de estas sustancias durante exposiciones laborales y domésticas y sobre su relación con la contaminación de las fuentes de agua y alimentos, colocando énfasis en la población de niños en edad escolar, debido a su mayor vulnerabilidad, reducción de la capacidad de desintoxicación y sus efectos a nivel endocrino y fisiológico.

Antioxidant and hypoglycemic effects of Momordica cochinchinensis Spreng: (Gac) aril extract on reproductive damages in streptozotocin (STZ)-induced hyperglycemia mice / Efectos antioxidantes e hipoglucémicos del extracto de aril de Momordica cochinchinensis Spreng: (Gac) sobre los daños reproductivos en ratones con hiperglucemia inducida por streptozotocin (STZ)

The aim of present study was to investigate the effect of Momordica cochinchinensis (Gag) aril (GA) aqueous extract on male reproductive system of streptozotocin (STZ)-induced hyperglycemia (HG) mice. GA were extracted with distilled water (DW) and analyzed for in vitro antioxidant capacities. ICR male mice were divided into 7 groups: 1) control, 2) DW, 3) GA 1000 mg/kg BW, 4) HG, 5) HG + glibenclamide, 6 and 7) HG + GA 500 and 1000 mg/kg BW respectively (7 mice/ group). In HG groups, mice were induced by STZ at single dose (150 mg/kg BW). They were treated for consecutive 35 days. All groups were compared for blood glucose levels, weights and histopathologies of reproductive organs, sperm concentration including testicular tyrosine phosphorylation protein patterns by Immuno-Western blotting. The results showed that GA processed antioxidant activities and could significantly decrease blood glucose levels and increase sperm concentration in HG mice. Moreover, GA could change the density of a testicular 70 kDa protein in HG-GA groups. In conclusion, GA extract could improve hyperglycemia and male reproductive damages in STZ-induced HG mice.

El objetivo de este estudio fue investigar el efecto del extracto acuoso de Momordica cochinchinensis (Gag) aril (GA) en el sistema reproductor masculino de ratones hiperglucémicos inducidos por estreptozotocina (STZ). GA fue extraída con agua destilada (DW) y se analizaron las capacidades antioxidantes in vitro. Ratones ICR machos fueron divididos en 7 grupos: 1) control, 2) DW, 3) GA 1000 mg / kg PC, 4) HG, 5) HG + glibenclamida, 6 y 7) HG + GA 500 y 1000 mg / kg PC, respectivamente (7 ratones / grupo). En los grupos HG, los ratones fueron inducidos con STZ en dosis única (150 mg / kg BW). Fueron tratados durante 35 días consecutivos. En todos los grupos se compararon los niveles de glucosa en sangre, los pesos y las histopatologías de los órganos reproductores, la concentración de espermatozoides, incluídos los patrones testiculares de proteínas tirosina fosforilada por Inmuno-Western blot. Los resultados mostraron que GA procesaba actividades antioxidantes y podían disminuir significativamente los niveles de glucosa en sangre y aumentar la concentración de espermatozoides en ratones HG. Además, GA podría cambiar la densidad de una proteína testicular de 70 kDa en grupos HG-GA. En conclusión, el extracto de GA podría mejorar la hiperglucemia y los daños reproductivos masculinos inducidos por STZ en ratones HG.

High exposure to boron in drinking water and sperm parameters in Chilean young people / Exposición elevada a boro en el agua potable y parámetros de esperma en gente joven chilena

Our aim was to describe sperm parameters in residents from Northern Chile. We evaluated in 101 volunteers (18 and 30 years old) urinary and drinking water Boron levels using Inductively Coupled Plasma Mass Spectrometry; semen parameters were measured with standardized methods. Each individual was categorized in 3 levels of exposure: low (B levels in urine 2.94 mgL-1 or tap water 3.0 mgL-1), medium (urinary B between 2.95-7.4 mgL-1 and B in tap water with 3.0-7.0 mgL-1) and high (urinary B > 7.4 mgL-1 or tap water > 7.0 mgL-1). We found no significant differences among groups by pH, sperm concentration (45.1; 48.2 and 38 million/mL), motility 1th hour (38.1; 40.0 and 45.5 %) and vitality 1th hour (88.6; 88.0 and 76.9 %) respectively. Abnormal morphology was significant different (83.3; 90 and 83 %). Young men exposed to B in drinking water present sperm variations associated with the level of exposure. Most of these changes are positive at intermediate levels of B. For the highest exposures were observed negative changes in sperm morphology, concentration, motility and vitality, all relevant parameters of fertility. Beneficial effect is observed at medium exposure, like a "U curve".

El objetivo de este trabajo fue describir los parámetros espermáticos en residentes del norte de Chile. Se evaluaron en 101 voluntarios (18 y 30 años), los niveles urinarios y de agua potable de boro, usando "Inductively Coupled Plasma Mass Spectrometry". Los parámetros del semen se midieron con métodos estandarizados. Cada individuo se clasificó en 3 niveles de exposición: bajo (niveles B en la orina 2,94 mgL-1 o agua potable 3,0 mgL-1), medio (B urinario entre 2,95-7,4 mgL-1 y B en agua de beber con 3,0- 7,0 mgL-1) y alto (B urinario >7,4 mgL-1 o agua potable > 7,0 mgL-1). No se encontraron diferencias significativas entre los grupos por pH, concentración de espermatozoides (45,1; 48,2 y 38 millones/mL), motilidad a 1 hora (38,1; 40,0 y 45,5%) y vitalidad 1 hora (88,6; 88,0 y 76,9%) respectivamente. La morfología anormal fue significativamente diferente (83,3; 90 y 83%). Los hombres jóvenes expuestos a B en el agua potable presentan variaciones espermáticas asociadas con el nivel de exposición. La mayoría de estos cambios son positivos en niveles intermedios de B. Para las exposiciones más altas se observaron cambios negativos en la morfología, concentración, motilidad y vitalidad del esperma, parámetros relevantes de la fertilidad. Un efecto beneficioso se observa en la exposición media, como una "curva U".

Combined protective effect of zinc oxide nanoparticles and melatonin on cyclophosphamide-induced toxicity in testicular histology and sperm parameters in adult Wistar rats

Background: Cyclophosphamide [CP] has been known as an anticancer drug with several side effects on various organs such as a male reproductive system that can cause infertility

Objective: To evaluate the possible combined effects of zinc oxide nanoparticles [nZno] and melatonin [Mel] on sperm parameters and histopathological changes of the testis in CP-treated rats

Materials and Methods: 42 adult male Wistar rats were divided into six groups. GI: control, GII: 60 mg/kg/wk CP, GIII and GIV, 10 mg/kg/wk Mel and 5mg/kg/wk nZno and GV: 5 mg/kg/wk nZno and 10 mg/kg/wk Mel were given 2 hr prior to CP injection, respectively, GVI: 5mg/kg/wk nZno and 10 mg/kg/wk Mel simultaneously. After 8 wk of treatment, rats were sacrificed and testis and epididymis were harvested for further evaluation

Results: The CP-treated group showed significant decreases in the body, testes and epididymis weights and sperm parameters [sperm count, viability, motility] with an increase abnormal sperms when compared with the control [p<0.001], as well as many histological alterations included decreased diameters of seminiferous tubules and Johnsen's Testicular Score [with degeneration, desquamation, multi-nucleated giant cell formation], whereas combined treatment [GV], showed more protective effects on CP-induced reproductive system damage compared with groups III or IV [p<0.001]

Conclusion: These results suggest simultaneous administration of Mel and nZno have more effectively protections against CP-induced reproductive damage than Mel or nZno alone

Wheat sprout effects on histological and histometrical structure and sperm parameters in testis of rat exposed to lead

Background: Wheat sprout contains a high amount of antioxidants, vitamins [especially vitamin E], minerals and phytoestrogen compounds. Use of medicinal herbs in reducing heavy metal toxicities has increased worldwide. In recent years, negative effects of lead on the male reproductive system and sperm fertility parameters have been shown broadly

Objectives: This study investigated the effects of wheat sprout extract [WSE] and vitamin E on sperm parameters and testicular oxidative stress in rats exposed to lead acetate

Methods: Thirty-five rats were divided randomly into seven groups: Gl [control group] received 1 ml/kg/day of normal saline, G2 received 20 mg/kg/day of lead acetate, G3 and G4 received 100 mg/kg/day and 200 mg/kg/day of WSE respectively, G5 and G6 received 100 mg/kg/day and 200 mg/kg/day of WSE respectively with 20 mg/kg/day of lead acetate, and G7 received 100 mg/kg/day of vitamin E with 20 mg/kg/day of lead acetate. After 35 days, rats were sacrificed and blood, sperm, liver and testicle tissue samples were collected for histomorphological and histochemical studies

Results: Results showed that count, motility and viability of sperms increased following the administration of 200 mg/kg/day of WSE [p<0.01]. Histomorphological studies showed a significant increase in tubular differentiation index [TDI], Repopulation index [RI], number of Sertoli cells, and epithelium of seminiferous tubules in groups receiving 200 mg/kg/day of WSE [p<0.00l]

Conclusions: Results of the current study show that dose dependent WSE significantly prevents testicular toxicity and oxidative stress effects of lead acetate