Efecto de la Vitamina E en Células Peritubulares de Testículos de Ratones Mus musculus Tratados con Ácido Valproico Durante el Estado Fetal y Pubertad / Effect of Vitamin E on Peritubular Cells from Testis of Mus musculus Mice Treated with Valproic Acid During the Fetal Stage and Puberty

El ácido valproico (VPA) es un fármaco antiepiléptico teratógenico que, al ser administrado durante etapas tempranas del embarazo, puede producir alteraciones en el desarrollo embriofetal, las que se manifiestan tanto a nivel del sistema nervioso como del testículo. No obstante, se ha reportado que la administración de vitamina E (VE) podría revertir dichas alteraciones. El objetivo del presente estudio fue determinar el efecto protector de la VE a nivel testicular en fetos y ratones púberes expuestos a VPA durante la fase embrionaria de su desarrollo. Se utilizó un total de 30 ratones hembra adultas gestantes (Mus musculus) cepa BALB/c, las cuales se dividieron en 6 grupos. El estudio contempló el análisis de fetos machos a los 17,5 días post-coital (dpc) y machos juveniles a las 6 semanas post-natal. A los grupos 1 y 4 se les administró 0,3 mL de solución fisiológica (grupos control para 17,5 dpc y 6 semanas postnatal, respectivamente). A los grupos 2 y 5 se les suministró la cantidad de 600 mg/kg de VPA (grupos VPA), en tanto que a los grupos 3 y 6 se les aplicó la misma dosis de VPA complementada con 200 UI de VE (grupos VPA+VE). Se describió la histología normal y patológica del compartimento peritubular del testículo. En los grupos VPA se evidenció una degeneración de la pared peritubular, y atrofia de túbulos seminíferos, así como exfoliación de las células germinales. Por el contrario, en los grupos VPA+VE tales signos no fueron observados y la morfología presentó aspecto normal solo con algunas alteraciones focales. Estos resultados corroboran el hecho que la administración de VE contrarresta en parte, los efectos deletéreos que ocasiona el VPA.

SUMMARY: Valproic acid (VPA) is a teratogenic antiepileptic drug that, when administered during the early stages of pregnancy, can produce alterations in embryo-fetal development, which manifest both at the level of the nervous system and the testicle. However, it has been reported that the administration of vitamin E (VE) could reverse these alterations. The study aimed to determine the protective effect of VE at the testicular level in fetuses and pubertal mice exposed to VPA during the embryonic phase of their development. 30 pregnant adult female mice (Mus musculus) BALB/c strain were used, which were divided into 6 groups. The study included the analysis of male fetuses at 17.5 days post-coital (dpc) and juvenile males at 6 weeks post-natal. Groups 1 and 4 were administered 0.3 mL of physiological solution. Groups 2 and 5 were given 600 mg/kg of VPA (VPA groups), while groups 3 and 6 were given the same dose of VPA supplemented with 200 IU of VE (VPA+VE). The normal and pathological histology of the peritubular compartment of the testis was described. In the VPA groups, degeneration of the peritubular wall, and atrophy of the seminiferous tubules, as well as exfoliation of the germ cells, were evident. On the contrary, in the VPA+VE groups such signs were not observed and the morphology presented a normal appearance with only some focal alterations. These results corroborate the fact that the administration of VE partially counteracts the deleterious effects caused by VPA.

Xiaokang Liuwei Dihuang decoction ameliorates the immune infertility of male rats induced by lipopolysaccharide through regulating the levels of sex hormones, reactive oxygen species, pro-apoptotic and immune factors.

Male immune infertility is a kind of disease that damages family life and happiness. The development of novel methods treating male immune infertility is of great significance. This study aimed to investigate the therapeutic effects of Chinese medicine Xiaokang Liuwei Dihuang decoction on immune infertility of male rats and explored the involved mechanisms. Model rats were established by lipopolysaccharide (LPS) injection. Anti-sperm antibody (AsAb) was detected by ELISA assay and testicular cell apoptosis was evaluated by TUNEL staining to verify the successful model establishment and screen suitable doses of Xiaokang Liuwei Dihuang decoction. Thirty rats were then divided into five groups (n = 6 per group): Control, LPS, Xiaokang Liuwei Dihuang decoction (15.12 g/kg, 30.24 g/kg and 45.36 g/kg). Results of HE staining showed that compared with LPS group, Xiaokang Liuwei Dihuang decoction treatments gradually improved the morphology of seminiferous tubules and elevated the number of spermatogenic cells as the doses increased. The sperm number and the levels of testosterone, luteinizing hormone (LH) and follicle stimulating hormone (FSH) in the serum of 15.12 g/kg, 30.24 g/kg and 45.36 g/kg Xiaokang Liuwei Dihuang decoction groups were much higher than those in LPS group. Results of TUNEL staining, ELISA assay and western blot showed that compared with LPS group, the testicular cell apoptosis and the levels of interleukin 1ß (IL-1ß), tumor necrosis factor α (TNF-α), AsAb, malondialdehyde (MDA) and toll-like receptor 2 (TLR2) in the testicular tissue significantly decreased in three Xiaokang Liuwei Dihuang decoction groups. Compared with LPS group, Bax expression in the 30.24 g/kg and 45.36 g/kg Xiaokang Liuwei Dihuang decoction groups was significantly down-regulated as well. In conclusion, Xiaokang Liuwei Dihuang decoction might ameliorate the immune infertility of male rats induced by LPS through regulating the levels of sex hormones, reactive oxygen species, pro-apoptotic and immune factors.

Effects of Corchorus olitorius and protacatechuic acid on diabetic rat testis tissue / Efectos de Corchorus olitorius y ácido protocatéquico en el tejido testicular de rata diabética

SUMMARY: The aim of this study is to investigate the effects of Protocatechuic acid and Corchorus olitorius on streptozotocin (STZ) induced diabetic rat testis tissue. Randomly selected Wistar Albino rats were divided into five groups as; Diabetes Mellitus, Diabetes Mellitus treated with Corchorus Olitorus (STZ+CO), Diabetes Mellitus treated with Protacatechuic acid (STZ+PCA), Corchorus olitorius (CO), Protocatechuic acid (PCA) and Control. Diabetic model was generated by intraperitoneal injection of 60 mg/kg Streptozotosin. After 48 hours of the STZ injection, blood samples were collected from tail vein in order to measure blood glycose levels. Over 250 mg/dL accepted as diabetic subjets and fed with 250 mg/kg Corchorus olitorius or 20 mg/kg PCA by oral gavage for three weeks. At the end of the experiment, right testes were removed and fixed in 10 % neutral formaldehyde for paraffine embedding. Sections were stained by HE, Masson trichrome, PAS and TUNEL for microscopic evaluation. Control, PCA-only and Corchorus olitorius-only treated group testes tissues showed a normal tissue organization, when degeneration in seminiferous tubules, the vacuolization, seperations in spermatogenic cell series, outpouring of cell groups in the lumen, vesicular body formation, liquid accumulation in the interstitial region and edema were observed in STZ induced diabetic models and untreated groups. Besides, higher amount of TUNEL (+) stained cells were determined in STZ group. On the other hand, blood glucose level and number of TUNEL (+) stained cells were decreased as a result of PCA and Corchorus olitorius treatment. Because of the reduction of blood glucose level and apoptotic cell numbers, PCA and Corchorus olitorius decreace the complications of diabetes mellitus induced rat testis.

RESUMEN: El objetivo de este estudio fue investigar los efectos del ácido protocatéquico y Corchorus olitorius sobre el tejido testicular de rata diabética inducida por estreptozotocina (STZ). Las ratas Wistar Albino fueron seleccionadas al azar y se dividieron en cinco grupos; Diabetes Mellitus, Diabetes Mellitus tratada con Corchorus olitorius (STZ + CO), Diabetes Mellitus tratada con ácido protocatéquico (STZ + PCA), Corchorus olitorius (CO), ácido protocatéquico (PCA) y Control. El modelo diabético se generó por inyección intraperitoneal de 60 mg/kg de estreptozotosina. Después de 48 horas de la inyección de STZ, se recogieron muestras de sangre de la vena de la cola para medir los niveles de glucosa. Niveles mayores a 250 mg/dL fueron considerados como especímenes diabéticos y alimentados con Corchorus olitorius de 250 mg/kg o PCA de 20 mg/kg por sonda oral durante tres semanas. Al final del experimento, se extirparon los testículos derechos y se fijaron en formaldehído neutro al 10 % para la inclusión en parafina. Las secciones se tiñeron con HE, tricromo de Masson, PAS y TUNEL para evaluación microscópica. Los tejidos de los testículos de los grupos control, tratados solo con PCA y con Corchorus olitorius mostraron una organización tisular normal. En cambio en modelos diabéticos inducidos por STZ y grupos no tratados se observó degeneración en los túbulos seminíferos, vacuolización, separaciones en series de células espermatogénicas, efusión de grupos celulares en la luz, formación del cuerpo vesicular, acumulación de líquido en la región intersticial y edema. Además, se determinó una mayor cantidad de células teñidas con TUNEL (+) en el grupo STZ. Por otro lado, el nivel de glucosa en sangre y el número de células teñidas con TUNEL (+) disminuyeron como resultado del tratamiento con PCA y Corchorus olitorius. Debido a la reducción del nivel de glucosa en sangre y el número de células apoptóticas, se observó que PCA y Corchorus olitorius disminuyen las complicaciones de los testículos de rata inducidos por diabetes mellitus.

Green tea consumption increases sperm concentration and viability in male rats and is safe for reproductive, liver and kidney health.

Green tea is a popularly consumed beverage worldwide and contains polyphenols, whose antioxidant activities could improve sperm parameters and fertility thereof. We investigated the effect of green tea on the male rat reproductive system as well as its safety. Male Wistar rats were administered 2 and 5% aqueous extract of green tea for 52 days' ad libitum, while the control group received tap water. Total polyphenol, flavanol, flavonol and soluble solids significantly increased in a concentration-dependent manner in vitro (P < 0.01). Weights of body, testis, epididymis, prostate gland, seminal vesicles, and liver, serum levels of testosterone, ferric reducing antioxidant power, creatinine, and sperm motility, remained unchanged (P > 0.05). Kidney weight, sperm concentration and vitality, spontaneous acrosome reaction increased (P < 0.05), while alanine transaminase and aspartate transaminase levels decreased (P < 0.05). Catalase, superoxide dismutase, glutathione and lipid peroxidation remained unchanged in the testes, liver and kidney (P > 0.05). Histological sections of testis, epididymis, kidney and liver showed no conspicuous alteration. Diameter and epithelial height of seminiferous tubule decreased, while caudal epididymis epithelial height increased (P < 0.01). Consumption of green tea in the conditions used in the present study seems to be safe and improved sperm parameters. However, subtle structural changes observed in the decreased diameter and epithelial height of the seminiferous tubule and increased acrosome reaction needs further investigation.

Deleterious effects of Pfaffia glomerata (Spreng.) Pedersen hydroalcoholic extract on the seminiferous epithelium of adult Balb/c mice.

Several plant species such as Pfaffia glomerata are widely used in traditional Brazilian medicine as stimulants and aphrodisiacs. In this regard, the aim of our study was to explore the effects of the long-term intake of the hydro-alcoholic root extract of P glomerata on the germ and somatic cells within the seminiferous tubules in adult Balb/c mice. The experimental groups were placed as: controls (water and DMSO), and treated with 300 and 400 mg/kg of the root extract. The number of germ and somatic cells, the proportion of pathological seminiferous tubules, and the germ cell apoptotic levels were evaluated. The volume and proportion of the seminiferous epithelium was decreased after the extract intake due to the increased germ cell apoptotic levels. Vacuolization of Sertoli cell cytoplasm was observed widely in pathological tubules, along with fully disorganized epithelia, showing multinucleated cells, which lead to decreased daily sperm production. Taken together, our results indicate that long-term intake of the P glomerata caused deleterious effects on spermatogenesis by inducing apoptosis and altering the seminiferous tubule's epithelial dynamics.

N-(3-Methozybenzyl)-(9Z,12Z,15Z)-octadecatrienamide from maca (Lepidium meyenii Walp.) ameliorates corticosterone-induced testicular toxicity in rats.

This study investigated the protective effects of maca ethanol extract (EEM) and N-(3-methozybenzyl)-(9Z,12Z,15Z)-octadecatrienamide (M 18:3) on corticosterone (CORT)-induced testicular toxicity. Male Wistar rats were divided into 5 groups. Except for the control group, CORT (40 mg per kg·bw) was injected subcutaneously for 21 consecutive days to induce testicular toxicity. 1 h before CORT injection, the rats were treated with EEM (400 mg per kg·bw) and M 18:3 (5 mg per kg·bw, 25 mg per kg·bw) by gavage, except for the control group and model group. Epididymal sperm and biochemical, and histological parameters were evaluated for the protective effects of the drugs. EEM (400 mg per kg·bw) and M 18:3 (5 mg per kg·bw, 25 mg per kg·bw) increased the sperm concentration and sperm motility, decreased the production of abnormal sperms, and increased the number of spermatogonia and primary spermatocytes in the seminiferous tubules of CORT-induced rats. Moreover, EEM and M 18:3 decreased the MDA levels and the positive expression rates of TUNEL, whereas they increased the activities of SOD, CAT, GSH-Px, and GST, and the contents of GSH in the testicles of CORT-induced rats. Furthermore, EEM and M 18:3 alleviated CORT-induced reduction in the positive expression rates of PCNA and Ki67 in the testicles of rats. Besides, EEM and M 18:3 reduced the expression levels of Keap-1 and increased the expression levels of Nrf2, HO-1, γ-GCS, and NQO1 in the testicles of CORT-induced rats. In summary, the protective effects of EEM and M 18:3 may be attributed to their anti-oxidative and anti-apoptotic properties.

Evaluation of male infertility treatment following Rhus coriaria extract administration on rats exposed to morphine.

Morphine is the most common analgesic drug that is widely used in post-operative interventions. This drug causes free radical accumulation leading to spermatogenesis failure. Antioxidant agents like Sumach (Rhus coriaria) neutralize cellular free radicals. In this study, the properties of antioxidative, modulative of inflammatory cytokines, and apoptotic genes following Sumach extract administration on morphine-induced fertility destruction in male Wistar rats was evaluated. Sixty-four animals were grouped (n = 8) including; 1: control, 2: morphine, 3-5: Sumach (200, 400, 800 mg/kg), and 6-8: morphine + Sumach. Hydroalcoholic extract of Sumach seeds was prepared. Treatments with Sumach extract were applied orally and intraperitoneally daily for 8 weeks. The P53, Bcl2 and caspase-3 genes expression were measured by real-time PCR. Cytokines involved in inflammation were evaluated by ELISA. Sperm parameters, total antioxidant capacity (TAC), testosterone, and germinal layer height (GLH) were assessed. All parameters (investigated in this study) in Morphine group reduced significantly than the control group (P ˂ 0.01) (except P53 and caspase-3 genes expression and inflammatory cytokine which were improved). All factors in Sumach and Sumach + Morphine groups were significantly enhanced compared to the Morphine group (P ˂ 0.01) (except P53 and caspase-3 genes expression and inflammatory cytokine which were declined). Morphine disrupted the physiological function of male fertility system. Besides, all doses of Sumach showed no therapeutic changes compared to the control group. Sumach with anti-infertility features compensates the toxic effect of Morphine administration.

Chestnut polysaccharides benefit spermatogenesis through improvement in the expression of important genes.

Recently there has been a continuing worldwide decrease in the quality of human spermatozoa, especially in spermatozoa motility and concentration. Many factors are involved in this decline, and great efforts have been made to rescue spermatogenesis; however, there has been little progress in the improvement of sperm quality. Chestnuts are used in traditional Chinese medicine; their major active components are chestnut polysaccharides (CPs). CPs have many biological activities but their effects on spermatogenesis are unknown. The current investigation was designed to explore the impact of CPs on spermatogenesis and the underlying mechanisms. We demonstrated that CPs significantly increased sperm motility and concentration (4-fold and 12-fold, respectively), and improved seminiferous tubule development by increasing the number of germ cells after busulfan treatment. CPs dramatically rescued the expression of important genes and proteins (STRA8, DAZL, SYCP1, SYCP3, TNP1 etc.) in spermatogenesis. Furthermore, CPs increased the levels of hormone synthesis proteins such as CYP17A1 and HSD17ß1. All the data suggested that CPs improved the testicular microenvironment to rescue spermatogenesis. With CPs being natural products, they may be an attractive alternative for treating infertile patients in the future. At the same time, the deep underlying mechanisms of their action need to be explored.

Can maternal exposure to paracetamol impair reproductive parameters of male rat offspring?

Paracetamol is a widely used medication during gestation and lactation periods for the treatment of pain and fever. Several studies have shown that exposure to paracetamol can increase the incidence of cryptorchidism and decrease testosterone production. Therefore, the present study aimed to evaluate if maternal treatment with paracetamol during gestation and gestation/lactation periods can alter reproductive and behavioral parameters in male offspring. Female Wistar rats were treated daily by gavage with water or paracetamol (350 mg/kg/day) during gestation (CTRG and PARG) or gestation/lactation periods (CTRGL and PARGL). There were significant differences in histomorphometry (increased volume and total length of the seminiferous tubules) and weight of testes (PARG group) and copulatory behavior and testosterone levels (PARG and PARGL groups) at PND 120. Therefore, the present study showed that maternal exposure to paracetamol has an impact on the reproductive system and sexual behavior of male adult offspring suggesting an impaired in sexual hypothalamic differentiation at the beginning of the development of the brain.

Vitamin E but Not GSH Decreases Reactive Oxygen Species Accumulation and Enhances Sperm Production during In Vitro Maturation of Frozen-Thawed Prepubertal Mouse Testicular Tissue.

Freezing-thawing procedures and in vitro culture conditions are considered as a source of stress associated with increased reactive oxygen species (ROS) generation, leading to a damaged cell aerobic metabolism and consequently to oxidative stress. In the present study, we sought to investigate whether vitamin E (Vit E) or reduced glutathione (GSH) enhances sperm production by decreasing ROS accumulation during in vitro maturation of prepubertal mice testes. Testes of prepubertal mice were cryopreserved using a freezing medium supplemented or not supplemented with Vit E and were cultured after thawing. In presence of Rol alone in culture medium, frozen-thawed (F-T) testicular tissues exhibited a higher ROS accumulation than fresh tissue during in vitro culture. However, Vit E supplementation in freezing, thawing, and culture media significantly decreased cytoplasmic ROS accumulation in F-T testicular tissue during in vitro maturation when compared with F-T testicular tissue cultured in the presence of Rol alone, whereas GSH supplementation in culture medium significantly increased ROS accumulation associated with cytolysis and tissue disintegration. Vit E but not GSH promoted a better in vitro sperm production and was a suitable ROS scavenger and effective molecule to improve the yield of in vitro spermatogenesis from F-T prepubertal mice testes. The prevention of oxidative stress in the cytoplasmic compartment should be regarded as a potential strategy for improving testicular tissue viability and functionality during the freeze-thaw procedure and in vitro maturation.

A biochemical and histological approach to study antifertility effects of methanol leaf extract of Asplenium dalhousiae Hook. in adult male rats.

The study was designed to investigate the antifertility properties of methanol leaf extract of Asplenium dalhousiae in adult male rats. Forty adult male Sprague Dawley rats (150 ± 10 g) divided into four groups (n = 10 animals/group) were administered with different doses (0, 50, 100, 150 mg/kg) of plant extract for 28 days. On day 29th, rats were decapitated, trunk blood and reproductive tissues were collected, and blood plasma was separated and stored until use for measuring reproductive hormones, while epididymis and testis were used for assessment of sperm parameters, oxidative stress status and morphometric analysis. Sperm motility, viability and sperm production rates were lowered in high dose treatment groups. Levels of catalase (CAT), sodium dismutase (SOD) and peroxidase (POD) decreased while stress biomarkers including reactive oxygen species (ROS) and thiobarbituric acid reactive substances (TBARS) were increased among all treatment groups. Concentrations of plasma testosterone and follicle stimulating hormone (FSH) were decreased while levels of luteinizing hormone (LH) increased in high extract treated groups. Histological examination of testis showed disorganisation of seminiferous tubule and reduced spermatocytes number. The findings of current study revealed that methanol leaf extract of A. dalhousiae might induce antifertility effects via oxidative stress and interfering with testicular architecture leading to spermatogenic arrest.

Evaluation of ameliorating effect of lycopene against testicular toxicity due to diethylnitrosamine using biochemical, spermatological and histopathological data.

The aim of this study was to evaluate the possible therapeutic or protective effects of lycopene on diethylnitrosamine (DEN)-induced testicular lipid peroxidation and on the associated changes in spermatological parameters and histopathological architecture of rat testis. DEN is a carcinogenic substance that can be derived from chemicals used in agriculture, such as insecticides and nitrate. The rats were assigned to control, lycopene, DEN(1), DEN(2), lycopene + DEN(1), lycopene + DEN(2), DEN(1) + lycopene and DEN(2) + lycopene groups. During the study, lycopene was administered by oral gavage at a dose of 10 mg kg-1  bw-1 every other day for 10 days and DEN was administered at a dose of 200 mg  kg-1  bw-1 as a single dose intraperitoneally. DEN was applied for 30 days in group DEN(1) and for 90 days in group DEN(2). Malondialdehyde (MDA) and reduced glutathione (GSH) levels, antioxidant enzymes activities, spermatological parameters, the weight of the reproductive organs (v. seminalis, prostate, testis and epididymis) and the histopathological structure were determined. MDA levels significantly increased, while GSH and antioxidant enzymes' activities decreased in DEN groups (p < 0.001). There was an increase in the rate of abnormal spermatozoa and a decrease in sperm density and motility, and reproductive organ weight (the weight of the right and left testis) in both DEN groups. Lycopene has normalised biochemical and spermatological parameters and reproductive organ weight. The histopathological examination of testicular tissue showed that the most significant histopathological change in DEN groups was the seminiferous tubule dilatation. These results suggest that besides the protective effects, the therapeutic effect of lycopene is possibly due to its antioxidant effects on DEN-induced testicular toxicity.

The effect of l-arginine and flaxseed on plasma testosterone concentration, semen quality and some testicular histology parameters in old broiler breeder roosters.

The objective of this study was to investigate the effect of dietary supplementation of l-arginine and flaxseed on testosterone and lipid parameters of serum and semen quality, as well as histological and morphometric parameters of testes in old broiler breeder rooster. Thirty six 52-week-old Ross 308 broiler breeder roosters with similar weights (4900 g ±â€¯210) were used for a 8-weeks trial period in a 3 × 3 factorial arrangement of dietary treatments that three levels of l-arginine (0.52, 0.68 and 0.83%) and flaxseed (0, 2 and 4%) with four replications in each. The ratio of left testicle weight/total testicular weight was significantly higher in A68F0 group. The maximum and minimum seminiferous tubule diameters were recorded for roosters fed no flaxseed and F2 flaxseed, respectively. The roosters fed A68 showed the highest serum testosterone. Seminal volume and sperm concentration, were not significantly affected by l-arginine × flaxseed interaction; however, sperm forward motility were significantly affected by arginine × flaxseed interaction (P < 0.05). A68F0 had significantly greatest tubular differentiation indices (TDI) in all of the treatments. Also, repopulation indices (RI) and spermiogenesis indices (SI) in A68F0 were significantly higher than other groups, but A52F0 and A52F4. However, testis index, testis density, serum lipids (cholesterol, LDL, HDL and HDL: LDL ratio), epithelium height of seminiferous tubules and number of Sertoli cells were not affected by the treatments. It is concluded that dietary supplementation of arginine (0.68%) has positive effects on blood testosterone, semen quality and spermatogenesis index of aged roosters.

Histomorphometric evaluation of seminiferous tubules and stereological assessment of germ cells in testes following administration of aqueous leaf-extract of Lawsonia inermis on aluminium-induced oxidative stress in adult Wistar rats.

OBJECTIVES: This study aimed to investigate the 'Cytoprotective effect of Lawsonia inermis aqueous leaf-extract on aluminium-induced Oxidative stress in Histomorphometric of the Seminiferous tubule and Stereology of Germ Cells of adult male Wistar rats', assessing its effect on the Histomorphometry of the Seminiferous tubule and Stereology of Germ Cells. METHODS: Thirty-five adult male Wistar rats, weighing between 100-196g, and fifteen mice of the same weight range were used. Lawsonia inermis extracts and aluminum chloride (AlCl3) were administered for a period of three (3) weeks, with Five (5) rats per group. Group 1 (control), received rat pellets and distilled water. Group 2 received 60mg/kg/d aqueous extract. Group 3 received 0.5mg/kg/d of AlCl3. Group 4 received 0.5mg/kg/d of AlCl3 and 60mg/kg/d of aqueous extract orally. Group 5 received 0.5mg/kg/d of AlCl3 and 75mg/kg/d of aqueous extract orally. Group 6 received 0.5mg/kg/d of AlCl3 and 100mg/kg/d of aqueous extract orally. Group 7 received 0.5mg/k/d of AlCl3 and 5mg/Kg/d of ascorbic acid orally. Twenty-four hours after the last administration, the animals were weighed, sedated with chloroform and blood was collected. The testes were removed and weighed. RESULTS: There were statistically significant changes in the percentage of seminiferous tubular and seminiferous ductal diameter within the experimental animals in all the groups (p<0.05). Stereological findings revealed increase in spermatogonia, primary spermatocytes, round Spermatids and elongated spematids, spermatozoa, Sertoli cells population of the control rats while the rats given 0.5mg of aluminum chloride per kg of body weight had the lowest value (p<0.05). CONCLUSION: In this study, we demonstrated the affected histomorphometry of the seminiferous tubule and stereology of germ cells in testes, where stress impacts were most felt and subsequently translated into drastic reproductive dysfunction and distortion of spermatogenesis.

Tribulus terrestris Protects against Male Reproductive Damage Induced by Cyclophosphamide in Mice.

Tribulus terrestris (TT) has been considered as a potential stimulator of testosterone production, which has been related with steroidal saponins prevailing in this plant. Cyclophosphamide (CP) is the most commonly used anticancer and immunosuppressant drug, which causes several toxic effects, especially on the reproductive system. Patients who need to use CP therapy exhibit reduced fertility or infertility, which impacts both physically and emotionally on the decision to use this drug, especially among young men. We hypothesized that the treatment with TT dry extract would protect the male reproductive system against CP toxicity. Mice received dry extract of TT (11 mg/kg) or vehicle by gavage for 14 days. Saline or CP was injected intraperitoneally at a single dose (100 mg/kg) on the 14th day. Animals were euthanized 24 h after CP administration, and testes and epididymis were removed for biochemical and histopathological analysis and sperm evaluation. The dry extract of TT was evaluated by HPLC analysis and demonstrated the presence of protodioscin (1.48%, w/w). CP exposure increased lipid peroxidation, reactive species, and protein carbonylation and altered antioxidant enzymes (SOD, CAT, GPx, GST, and GR). Moreover, acute exposure to CP caused a reduction on 17 ß-HSD activity, which may be related to the reduction in serum testosterone levels, histopathological changes observed in the testes, and the quality of the semen. The present study highlighted the role of TT dry extract to ameliorate the alterations induced by CP administration in mice testes, probably due to the presence of protodioscin.

[Lipoic acid protects spermatogenesis in male rats with ornidazole-induced oligoasthenozoospermia].

OBJECTIVE: To study the protective effect of lipoic acid (LA) on the spermatogenic function of the male rats with oligoasthenozoospermia induced by ornidazole (ORN). METHODS: Seventy male SD rats were equally randomized into groups A (solvent control: 1 ml 0.5% CMC-Na + 1 ml olive oil), B (low-dose ORN model: 400 mg/kg ORN suspension + 1 ml olive oil), C (low-dose ORN + low-dose LA treatment: 400 mg/kg ORN + 50 mg/kg LA), D (low-dose ORN + high-dose LA treatment: 400 mg/kg ORN + 100 mg/kg LA), E (high-dose ORN model: 800 mg/kg ORN suspension + 1 ml olive oil), F (high-dose ORN + low-dose LA treatment: 800 mg/kg ORN + 50 mg/kg LA), and G (high-dose ORN + high-dose LA treatment: 800 mg/kg ORN + 100 mg/kg LA), and treated respectively for 20 successive days. Then all the rats were sacrificed and the weights of the body, testis, epididymis and seminal vesicle obtained, followed by calculation of the organ index, determination of epididymal sperm concentration and motility, and observation of the histomorphological changes in the testis and epididymis by HE staining. RESULTS: Compared with group A, group E showed significantly decreased body weight (ï¼»117.67 ± 11.53ï¼½ vs ï¼»88.11 ± 12.65ï¼½ g, P < 0.01) and indexes of the testis (ï¼»1.06 ± 0.12ï¼½ vs ï¼»0.65 ± 0.13ï¼½ %, P < 0.01) and epididymis (ï¼»0.21 ± 0.03ï¼½ vs ï¼»0.17 ± 0.01ï¼½ %, P < 0.01). In comparison with group E, group F exhibited remarkable increases in the epididymal index (ï¼»0.17 ± 0.01ï¼½ vs ï¼»0.20 ± 0.02ï¼½ %, P < 0.01), and so did group G in the body weight (ï¼»88.11 ± 12.65ï¼½ vs ï¼»102.70 ± 16.10ï¼½ g, P < 0.05) and the indexes of the testis (ï¼»0.65 ± 0.13ï¼½ vs ï¼»0.95 ± 0.06ï¼½ %, P < 0.01) and epididymis (ï¼»0.17 ± 0.01ï¼½ vs ï¼»0.19 ± 0.02ï¼½ %, P < 0.05), but no obvious difference was observed in the index of seminal vesicle among different groups. Compared with group A, group B manifested significant decreases in sperm motility (ï¼»74.12 ± 8.73ï¼½ vs ï¼»40.25 ± 6.08ï¼½ %, P < 0.01), and so did group E in sperm count (ï¼»38.59 ± 6.40ï¼½ vs ï¼»18.67 ± 4.59ï¼½ ×105/100 mg, P < 0.01) and sperm motility (ï¼»74.12 ± 8.73ï¼½ vs ï¼»27.58 ± 8.43ï¼½ %, P < 0.01). Sperm motility was significantly lower in group B than in C and D (ï¼»40.25 ± 6.08ï¼½ vs ï¼»58.13 ± 7.62ï¼½ and ï¼»76.04 ± 8.44ï¼½%, P < 0.01), and so were sperm count and motility in group E than in F and G (ï¼»18.67 ± 4.59ï¼½ vs ï¼»25.63 ± 9.66ï¼½ and ï¼»29.92 ± 4.15ï¼½ ×105/100 mg, P < 0.05 and P < 0.01; ï¼»27.58 ± 8.43ï¼½ vs ï¼»36.56 ± 11.08ï¼½ and ï¼»45.05 ± 9.59ï¼½ %, P < 0.05 and P < 0.01). There were no obvious changes in the histomorphology of the testis and epididymis in groups A, B, C and D. Compared with group A, group E showed necrotic and exfoliated spermatogenic cells with unclear layers and disorderly arrangement in the seminiferous tubules and remarkably reduced sperm count with lots of noncellular components in the epididymal cavity, while groups F and G exhibited increased sperm count in the seminiferous tubules and epididymis lumen, also with exfoliation, unclear layers and disorderly arrangement of spermatogenic cells, but significantly better than in group E. CONCLUSIONS: LA can reduce ORN-induced damage to the spermatogenetic function of rats, improve sperm quality, and protect the reproductive system.

Effect of Lead Acetate on Basement Membrane of Seminiferous Tubules of Adult Rat Testis and Protective Effects of Ficus Carica: A Histological Study.

OBJECTIVE: To determine the effects of lead acetate and Ficus carica on disruption of basement membrane in seminiferous tubules of adult rat testis. STUDY DESIGN: An experimental study. PLACE AND DURATION OF STUDY: Department of Anatomy, Army Medical College, Rawalpindi in collaboration with National Institute of Health (NIH), Islamabad, from March to November 2017. METHODOLOGY: Thirty male adult Sprague Dawley rats were selected and divided into three groups, each with ten animals. All treatments were given once daily for a period of eight weeks. Control was labelled as group A. Group B was administered lead acetate at a dose of 30 mg/kg body weight. Group C was treated with lead acetate at a dose of 30 mg/kg body weight and Ficus carica at a dose of 80 mg/kg body weight. Animals were dissected 24 hours after the last dose. Testis were treated, fixed and stained for histological study. Disruption of basement membrane in seminiferous tubules was scored morphometrically on a scale of 0 (normal) to 3 (>70% tubules showing disruption) and statistically analysed. RESULTS: Significant number of seminiferous tubules showed disruption of basement membrane in group B (20%) as compared to group A (0%). Less severe disruption of membrane was seen in group C as compared to group B, which was statistically not significant (p=0.082). CONCLUSION: Lead acetate causes significant disruption of basement membrane in seminiferous tubules of testis of adult rats but subsequent administration of Ficus carica reduces the effects on short term.

Protective effect of melatonin versus montelukast in cisplatin-induced seminiferous tubule damage in rats.

We compared the protective effects of melatonin and montelukast against cisplatin-induced testicular damage. Adult male rats were assigned to one of four groups: a control group, a cisplatin (Cis) group treated with a single intraperitoneal injection of 7 mg/kg cisplatin, a cisplatin + melatonin group (Cis-Mel) and a cisplatin + montelukast group (Cis-Mon) each treated with the same dose of cisplatin together with either oral melatonin (20 mg/kg) or oral montelukast (10 mg/kg) in 2 ml water from day 1 to day 10 starting on the day of the cisplatin injection. Cisplatin-induced oxidative stress, with a significant increase in testicular malonedialdehyde (MDA), decreased testicular glutathione (GSH), histological testicular damage and body weight loss. Additionally, increased abnormal sperm forms and decreased count and motility were noted. Melatonin and montelukast both rescued GSH concentrations, increased sperm count and motility and decreased abnormal forms. Montelukast resulted in better rescue of weight loss, while greater improvement in sperm count and testicular pathology, and a trend for decreased MDA were noted with melatonin. These findings suggest that melatonin and montelukast protect against different aspects of cisplatin-induced toxicity. Future studies should assess whether both drugs may have additive benefit when used in combination.

Protective effects of the hydroalcoholic extract of Fumaria parviflora on testicular injury induced by torsion/detorsion in adult rats.

This study was designed to determine the effects of daily oral administration (250 mg/kg) of the hydroalcoholic extract of Fumaria parviflora (FP) for 14 days on the sperm parameters, oxidative stress parameters, serum testosterone levels, expression of Bax and Bcl-2 genes, and apoptosis index of germ cells after testicular torsion-detorsion (ischaemia-reperfusion, IR) injury model in rats. Twenty-eight adult male Wistar rats were divided randomly into four groups of seven each: sham operation, torsion-detorsion (TD), TD plus the hydroalcoholic extract FP (TDFP) and only FP without TD application (FP). Testicular torsion was created by rotating the left testis 720° in a counterclockwise direction; then, after 4 hr, detorsion was performed. The Johnson's score, mean seminiferous tubule diameter (MSTD) and height (thickness) of seminiferous tubule epithelium (HST) were significantly increased in TDFP and FP groups as compared to TD group. The gene expression of Bcl-2, level of serum testosterone hormone and antioxidant parameters-GPx and SOD-were significantly higher in TDFP and FP groups than TD group. The index of apoptosis, the gene expression of Bax and the level of MDA were significantly higher in TD group than TDFP and FP groups. Therefore, F. parviflora could decrease oxidative stress induced by testicular torsion-detorsion.

Oxidative stress burden inhibits spermatogenesis in adult male rats: testosterone protective effect.

In this study, we aimed to investigate the protective effects of androgens, using letrozole (LET; an aromatase inhibitor), grape seed extract (GSE; a naturally occurring aromatase inhibitor and antioxidant), and testosterone propionate (Tp), against methotrexate (MTX)-induced testicular toxicity in adult male rats. MTX has been shown to induce oxidative stress and exhibit antiproliferative effects in the testes. Adult male rats received oral saline gavage (control group with no treatment), the potential protective agents (LET, GSE, or Tp) alone, MTX alone, or a combination of one of the potential protective agents and MTX. The testicular levels of oxidative stress markers and cytokines (tumor necrosis factor-α and interleukin-1ß) were measured. Spermatogenesis and sperm viability were microscopically evaluated. Administration of LET and GSE 7 days before MTX improved spermatogenesis and sperm viability, as well as reduced the levels of oxidative stress markers and cellular cytokines. Exogenous testosterone exhibited anti-inflammatory and antioxidant activities, similar to GSE and LET. We also showed that enhancing the endogenous androgenic activity by LET and GSE protected spermatogenesis against MTX-induced testicular toxicity via reduction of inflammation and oxidative stress in the testes. Our data suggest that testosterone protected spermatogenesis owing to its antioxidant and anti-inflammatory properties.