Objective(s): Increased levels of nitric oxide (NO) in the testicular veins of people suffering from varicocele have already been reported. However, the role of NO-synthase (NOS) isozymes and their inhibitors have not been extensively studied. We aimed to evaluate the inhibitory effects of aminoguanidine (AG), on sperm motility, vitality, and mitochondrial membrane potential (MMP) in varicocelized rats. Materials and Methods: Twenty fore male Wister rats were divided into control, sham, varicocele, and treatment groups. Varicocele and treatment groups underwent partial ligation of left renal vein. Rats in the sham group underwent the same procedures as the varicocele group with the exception of vein ligation. 10 weeks after varicocele induction, sperm parameters were evaluated in all groups. The treatment group received 50 mg/kg AG injection daily for 10 weeks after which they were sacrificed prior to assessment of the parameters. Sperm viability and MMP were assessed by flow cytometry using propidium iodide (PI) and rhodamine 123 (Rh123), respectively. Results: The results of this study show a decrease in sperm viability, motility and MMP in the varicocele group compared with the other groups. After AG injection, we observed that all the parameters were significantly enhanced in the treatment group compared with the other groups. Rh123 staining revealed a positive relation between MMP and sperm motility, whereas PI staining showed a positive relation between sperm motility and viability. Conclusion: The findings of our study show that AG improves sperm motility and MMP, and thus, might be useful in the management of varicocele-related infertility.

Background: Unique omega-3 fatty acids may contribute to the membrane fluidity which is necessary for the motility of sperm tails and also dietary fish oil (FO) can improve sperm quality. The positive effects of L-carnitine (LCAR) on male reproductive performance have been reported in some species. This study has been focused on the plasma membrane integrity and mitochondrial membrane potential in Iranian ram fed FO and/or LCAR, that till now, there is no report this issue.Materials and Methods: Nineteen Zandi rams were randomly assigned to experimental groups and were offered an isoenergetic and isonitrogenous ration. The treatments consisted of control (CTR; n=4); fish oil (FO); 35 g/d/ram; n=4); L-carnitine (LCAR; 220 ppm/d/ram; n=4) and FO+LCAR (n=4) with constant level of Vitamin E. In addition, three rams were used without Vitamin E.Diets were fed to the rams for 70 days and semen was collected once a week. Sperm routine parameters were analyzed by CASA. Sperm plasma membrane integrity and mitochondrial membrane potential (MMP) assessed by hypo-osmotic swelling test (HOST) and JC-1 respectively.Data were analyzed by SPSS 16.Results: The results showed that all sperm characteristics increased with FO. HOST-positive spermatozoa were significantly different between control and FO+LCAR (87 vs.82 vs.77 and 76% for FO+LCAR, FO, LCAR and CTR, respectively; p<0.05). MMP was affected and increased by inclusion FO+LCAR in diet (85.5 vs.76 and 75 vs.64%, FO+LCAR, FO, LCAR and CTR, respectively; p<0.05).Conclusion: Addition of FO in combination with LCAR could improve sperm functional test significantly.On the other hand, lonely LCAR at this supplemental level did not improve plasma membrane integrity in comparison with CTR, but it is effective on mitochondrial membrane potential. FO and LCAR treatments have similar effects on mitochondrial membrane potential.

Aim: This study was performed to investigate if silymarin can prevent the adverse effects of aluminum chloride on viability, motility and mitochondrial membrane potential in ram sperm. Material and Methods: Epididimal spermatozoa from Farahani's ram were divided into five groups: 1. Sperm at 0 hour, 2. Sperm at 180 minutes (control), 3. Sperm treated with aluminum chloride (0. 5 mM) for 180 minutes, 4. Sperm treated with silymarin (0. 5 μ M) + aluminum chloride (0. 5 mM) for 180 minutes and 5. Sperm treated with silymarin (0. 5 μ M) for 180 minutes. In order to evaluate viability and mitochondrial membrane potential in the groups, MTT ((3-4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltenyltetrazolium bromide) assay and Rhodamine 123 staining were used respectively. Sperm motility was done according to World Health Organization (WHO) protocol. Data were analysed using one-way ANOVA fallowed by Tukey's test. Results: The percentage of sperm viability, progressive motility and intact mitochondrial membrane potential were significantly decreased in the aluminum chloride group compared to the control. This pollutant also significantly increased the percentage of non-progressively motile and non-motile sperm compared to the control group. In silymarin + aluminum chloride group, silymarin could significantly compensate the adverse effect of aluminum chloride on the sperm parameters (except to non-progressively motile) compared to the aluminum chloride group. Conclusions: Aluminum chloride has toxic effect on ram sperm viability, motility and intact mitochondrial membrane potential and silymarin is able to compensate the adverse effect of aluminum chloride on these sperm parameters.

Background: Acrylamide (AA) is a compound used in the industrial production of polyacrylamide. AAs affects by creating oxidative stress. It produces reactive oxygen species and leads to lipid peroxide. Lipid peroxidation in the cell membrane is one of the most important oxidations in the sperm, which can disrupt the fluidity and permeability of cell membranes and damage all cells. Objective: To investigate the different concentrations of AA on human sperm parameters based on the World Health Organization standard and its impact on mitochondrial membrane potential and sperm glutathione levels. Materials and Methods: In this laboratory study, we examined the different concentrations of AA on human sperm parameters based on the World Health Organization standard and its impact on mitochondrial membrane potential by flow cytometry and sperm glutathione levels by ELISA assay. Results: The results were reported as the mean fluorescence intensity of JC and the index was observed to decrease following the effect of AA in mitochondrial membrane potential (Δ Ψ m). The results of ELISA test to study the level of intracellular glutathione showed that with the increase in the concentration of AA exposed to sperms, there was a significant reduction in the level of intracellular glutathione. Conclusion: AA destroys the sperm membrane integrity under apoptotic and oxidative inductions with a negative impact on mitochondrial function and antioxidative enzyme in sperm such as glutathione.

Cytotoxicity of depleted uranium, as a byproduct of military has been came to spotlight in recent decades. DU is known as a chemical rather than radioactive hazard and efforts to illustrating its mechanism is undergo, but the precise complete molecular mechanisms are still unclear. Recent studies showed that uranium induces biological changes in many different target tissues, such as the kidney, brain and skin. The aim of this study was to assess the impact of depleted uranium exposure at the cellular level in the human dermal fibroblast primary cells. The human dermal fibroblast primary cells incubated with different concentration (250-750 mM) of depleted uranium.Cytotoxicity and mitochondrial function in this cell lines were determined with the LDH leakage assay and the MTT test respectively. MDA levels were measured for determination of Lipid peroxidation in DU treated cells. Besides glutathione depletion and apoptosis phenotype detection were also assessed to complete the mechanistic screening. Results showed that the cell viability ameliorates in concentration and time dependent manners following in 24, 48and 72 h incubation with DU. Moreover the significant increase in lipid peroxidation and significant decrease in cellular GSH recorded in DU treated human dermal fibroblast primary cells suggesting the preoxidant effect of uranyl ions. Cytoprotective effects of N-acetylcysteine (NAC) and dramatic decrease of cell viability in buthionin sulfoxamid (BSO) pretreated cells indicated the possibility of a critical role for glutathione system in DU detoxification. Death pattern, in fibroblast cells following DU treatment was varied from apoptosis to necrosis while the time and concentration increased.Since ROS formation is the initiation step for cell apoptosis, the present studies suggestUranyl-induced toxicity in the human dermal fibroblast primary cells originated from oxidative stress and lead to occurrence of programmed cell death.

Background: Varicocele is a major cause of male infertility, but its pathophysiology is unclear. Recent studies declare that fertile varicocele people with normal semen analysis are also at risk of loss of infertility. The exact mechanism by which varicocele damages spermatogenesis is still unknown. Some studies have reported increased Reactive Oxygen Species (ROS) is a major factor in semen of men with varicocele. The aim of this study was to investigate whether the source of elevated ROS is intracellular or not. In addition, we studied Mitochondrial Membrane Potential (MMP), viability, antioxidant activity, sperm count and motility in these rats.Methods: The study group consisted of 28 male rats divided in four groups: control, sham, varicocele 1, varicocele 2, Experimental varicocele was established by partial ligation of the left renal vein in last two groups. Animals were sacrificed two and six months after surgery and dilation of the internal spermatic veins was observed. Then, superoxide anion production and Mitochondrial Membrane Potential were evaluated by Flow cytometry sperm characteristics were evaluated by Flow cytometry. Sperm superoxide anion production was assessed by the dihydroethidium and Mitochondrial Membrane Potential with rhodamin 123.Results: Our results showed intracellular superoxide anion production significantly increased and Mitochondrial Membrane Potential, viability, sperm count and motility significantly decreased in rats with experimental left varicocele. However, there was no significant difference for seminal plasma antioxidant activity between all groups.Conclusions: Consequently, our findings suggest that one of the main sources of ROS production is intracellular and we must consider it in treatment.