The effects of caffeine on ovarian tissue in rats.

Caffeine consumption increases during early adulthood, which has adverse effects on the reproductive system. This study aimed to assess the impact of embryonic caffeine exposure on rat ovary in adulthood. Female Wistar rats (240-270 g) were divided into 5 groups (n = 7): experimental groups were exposed to 26, 45, 100, and 150 mg/kg of caffeine via drinking water during pregnancy and the control group only received drinking water. The ovaries of the offspring were taken out on days 7, 14, 28, 60, 90, and 120 of postnatal development, and then, they were fixed in 10% formaldehyde solution. Ovarian follicles were studied using stereological methods, and data were analyzed using one-way ANOVA followed by the Tukey test in SPSS software. A value of p < 0.05 was considered significant. The body weight, the weight of the ovaries, the ovarian volume, and the number of primordial follicles decreased significantly (p < 0.05) in 45 and 100 mg/kg, and (p < 0.001) in 150 mg/kg caffeine-treated groups at all stages of postnatal development. Significant decreases were observed in the number of primary and secondary follicles in 45 and 100 mg/kg (p < 0.05) and (p < 0.001) in 150 mg/kg caffeine-treated groups on days 7, 14, 28, and 60 compared to the control group. The number of Graafian follicles also decreased significantly (p < 0.001) in 45, 100, and 150 mg/kg caffeine-treated groups on days 14 and 28. Moreover, the mean volume of the oocyte in Graafian follicles reduced considerably in 45, 100, and 150 mg/kg caffeine-treated groups compared to other groups (p < 0.05). The thickness of the zona pellucida (ZP) in the secondary follicles (p < 0.02) and Graafian follicles (p < 0.05) showed a significant reduction in 100 and 150 mg/kg caffeine-treated groups on the 14th, 28th, and 60th days. In conclusion, high-dose caffeine consumption during gestation affects all stages of ovarian follicle development in rat offspring.

Protective effect of vitamin E on oxidative stress and sperm apoptosis in diabetic Mice.

BACKGROUND: Generation of free radicals and oxidative stress are a major contributor to diabetes. These factors lead to the development of diabetic testicles disorders. OBJECTIVE: In this study, the protective effect of vitamin E on functional disorders associated with diabetes induced oxidative stress in male reproductive systems has been investigated. MATERIALS AND METHODS: Thirty-three adult male Mice were divided into control, diabetic, and untreated diabetic groups. Streptozotocin was used to induce diabetes. In the treated group, vitamin E was given to the Mice intraperitoneally for 30 days. Then, animals were anesthetized and sacrificed. Animal testicles were isolated and homogenized in phosphate buffer and used for measuring sperm count, motility and survival of sperm, MDA concentration and antioxidant capacity (TAC). Apoptosis was also performed with the TUNEL test. RESULTS: The results of reduction (12.03 ± 98.11) TAC, MDA concentration (-28.5 ± 2.58), sperm motility (unstable sperma= 86.4 ± 7.48), sperm count (171.51), Sperm morphology (natural morphology= 49.69 ± 31.93) and abnormal morphology (9.77 ± 49.7) with increased oxidative damage. These changes were statistically significant in comparison with the control group for all variables other than MDA (p= 0.05). Treatment of vitamin E diabetic Mice improved the ability of antioxidants to prevent oxidative damage in the testicles, restore the sperm movement, and increase the number of normal sperm as well as TAC. The level of apoptosis in the treated group has decreased compared to the untreated group. CONCLUSION: Vitamin E protects the reproductive system against diabetes mellitus. Therefore, it was concluded that vitamin E may be a suitable agent for protecting the sperm and testicular parameters against undesirable effects of diabetes.

Effects of different doses of tamoxifen on the sperm parameters and chromatin quality in mice: An experimental model.

BACKGROUND: Tamoxifen (TX) is widely used for the treatment of male factor and idiopathic infertility. It has been shown that TX induces sperm production and so improves male fertility. OBJECTIVE: This study evaluated the effects of different doses of TX on the sperm parameters and chromatin quality in mice. MATERIALS AND METHODS: In this research, 24 male NMRI mice were divided into three groups including group A: control animal receiving vehicle; group B: the group receiving basal diet and TX 0.4 mg/kg/day; and group C: the group receiving basal diet and TX 0.6 mg/kg/day for 35 days. Thereafter, epididymal spermatozoa were analyzed for standard parameters and nuclear chromatin quality using Aniline Blue (AB) and Toluidine Blue (TB) staining. RESULTS: The results indicated that although the TX did not affect the sperm count, motility, and viability parameters, it could elevate the percentage of sperm cells with abnormal morphology and abnormal chromatin at both doses. In addition, in comparison with the control mice, a significant elevation was observed in spermatozoa with residual histones (assessed by AB staining) at high doses of TX. CONCLUSION: Our experimental data in mice suggested that the use of TX for treating male infertility might increase the rates of spermatozoa with abnormal chromatin in a dose-dependent manner.

Effects of Tamoxifen on DNA Integrity in Mice.

BACKGROUND: Tamoxifen (TX) is widely used to treat idiopathic infertility in men. Using TX has been shown to produce sperm in patients with oligospermia and azoospermia and improve male fertility. The aim of this study was to evaluate the effects of TX on DNA and chromatin quality in mice regarding the importance of chromatin quality and sperm DNA at all stages of reproduction. METHODS: 24 male NMRI mice were divided into 3 groups including dose 0.4 mg/kg/day that received basal diet and TX, dose 0.6 mg/kg/day that received basal diet and TX, and group 3 that received vehicle for 35 days as the control. After that, epididymal spermatozoa were analyzed for nuclear DNA quality. One-way ANOVA was performed with a Tukey test to compare sperm DNA fragmentation at different times. The p<0.05 was considered significant. RESULTS: The use of different doses of TX may have detrimental effects on sperm chromatin protamination and DNA integrity in mice. According to Acridine Orange (AO) staining, the rate of increased single-stranded DNA damage was observed at 0/6 mg/kg/day TX dose (p<0.05). CONCLUSION: The use of different studied TX doses in the animal sample was found to increase the amount of protamine deficiency and DNA defect in treated mice.

Stereological Method for Assessing the Effect of Vitamin C Administration on the Reduction of Acrylamide-induced Neurotoxicity.

INTRODUCTION: Acrylamide (ACR) consumption is increasing all over the world. There are some evidence on the literature about its neurotoxic effect on mature animals, but the effects of ACR on postnatal development have been less studied. The purpose of this study was to evaluate the effects of ACR on development of cortical layer, white matter, and number of Purkinje cells of the cerebellum in rat newborns. METHODS: This study was carried out on 20 female Wistar rats (average weight: 180 g, aged: two months). The rats were divided into four groups. Pregnant rats were orally fed with ACR 10 mg/kg and vitamin C 200 mg/kg. In this study, 6 infants of each group (weighting 32-35 g) were randomly selected at day 21 after birth and placed under deep anesthesia and transcardial perfusion. Their cerebellums were fixed and histopathological changes were evaluated with Hematoxylin and Eosin (H&E) staining and cresyl violet method. The volume of cerebellar cortical layers and number of Purkinje cells were investigated by Cavalieri's principle and physical dissector methods. The obtained data were analyzed by 1-way ANOVA and LSD test using SPSS. P<0.05 considered as statistically significant. RESULTS: The results showed that newborns of ACR-treated female rats have decreased cerebellar weight (P≤0.05) and lower than average number of Purkinje cells (P≤0.001). ACR also decreased the volume of granular and molecular layer and increased the volume of white matter. While the results showed decreased in white matter volume in vitamin C group (P≤0.001). CONCLUSION: ACR induces structural changes in the development of the cerebellar cortical layers in rat newborns, but these changes may be prevented by vitamin C as an antioxidant.

Saccharin consumption increases sperm DNA fragmentation and apoptosis in mice.

BACKGROUND: Saccharin is an artificial non-caloric sweetener that used to sweeten products such as drinks, candies, medicines, and toothpaste, but our bodies cannot metabolize it. Sodium saccharin is considered as an important factor in tumor promotion in male rats but not in humans. OBJECTIVE: The objective of this study was to investigate the effect of saccharin consumption on sperm parameters and apoptosis in adult mice. MATERIALS AND METHODS: Totally 14 adult male mice were divided into 2 groups. Group 1 served as control fed on basal diet and group 2 or experimental animals received distilled water containing saccharin (0.2% w/v) for 35 days. After that, the left cauda epididymis of each mouse was cut and placed in Ham's F10. Swimmed-out spermatozoa were used to analyze count, motility, morphology (Pap-staining) and viability (eosin-Y staining). Sperm DNA integrity, as an indicator of apoptosis, was assessed by SCD (sperm chromatin dispersion) and terminal deoxynucleotidyl transferase (TUNEL) assay. RESULTS: Following saccharin consumption, we had a reduction in sperm motility with respect to control animals (p=0.000). In addition, the sperm count diminished (17.70±1.11 in controls vs. 12.80±2.79 in case group, p=0.003) and the rate of sperm normal morphology decreased from 77.00±6.40 in control animals into 63.85±6.81 in saccharin-treated mice (p=0.001). Also, we saw a statistically significant increase in rates of sperm DNA damage and apoptosis in experimental group when compared to control one (p=0.001, p=0.002 respectively). CONCLUSION: Saccharin consumption may have negative effects on sperm parameters, and increases the rate of sperm DNA fragmentation and apoptosis in mice.