Evaluation of the effect of mitoquinone on functional parameters, DNA structure, and genes expression related to the apoptotic and antioxidants of human sperm after freezing-thawing.

OBJECTIVE: Sperm freezing is considered as an effective way in assisted reproductive technology (ART) programs, it has detrimental effects on sperm function, due to the production of reactive oxygen species (ROS). This study aimed to investigate the potential of Mitoquinone (MitoQ) in inhibiting the production of mitochondrial ROS during sperm freezing. METHODS: A total of 20 human normozoosperm samples were collected for this study. The samples were divided into four groups, each containing different concentrations of MitoQ (0, 0.2, 2, and 20 nM), and then subjected to the freezing process. After thawing, the sperm suspensions were evaluated for parameters including motility, morphology, acrosome integrity, adenosine triphosphate (ATP) level, intracellular ROS, viability, chromatin packaging, DNA denaturation, DNA fragmentation, as well as the expression of antioxidants (GPX, SOD) and apoptotic (Bax, Bcl2) genes. RESULTS: The results showed that total and progressive mobility of sperms significantly increased in the 2 nM group, while significantly decreased in the 20 nM group (p ≤ 0.05). Sperm morphology did not significantly improve across all the tested concentrations (p ≥ 0.05). Intracellular ROS levels showed a significant decrease and increase in the concentrations of 2 and 20 nM, respectively (p ≤ 0.05). Furthermore, a significant increase was observed in viability, ATP, acrosome integrity, chromatin packaging, and non-denatured and non-fragmented DNA after treatment with 2 nM of MitoQ, compared with the control group (p ≤ 0.05). Regarding gene expressions, the relative expressions of oxidative stress genes were increased in the 2 nM group and decreased in the 20 nM group (p ≤ 0.05), while no significant difference was observed in the expressions of apoptotic genes compared with the control group (p ≥ 0.05). All the comparisons were made with respect to the control group. CONCLUSION: Adding the optimal concentration of MitoQ (2 nM) to the sperm freezing medium not only improves sperm functional parameters and reduces DNA damages, but also stimulates the expression of antioxidant genes, leading to even greater benefits for sperm cryopreservation.

Effects of BCc1 nanoparticle and its mixture with doxorubicin on survival of murine 4T1 tumor model.

Background: Our previous findings showed that BCc1, a nanoparticle designed based on nanochelating technology, can be considered a new anti-cancer nanoparticle if confirmed by complementary studies. Goal: In the present study, we investigated the effects of the BCc1 nanoparticle alone on some gene expressions influencing the apoptosis pathway, and also the effect of the mixture of BCc1 nanoparticle and doxorubicin on survival. Method: Using an in vitro study, the effects of the BCc1 nanoparticle on Bax, Bcl2, p53, Caspase7 and p21 gene expressions were assessed after a 24-h treatment using real-time PCR in MCF-7 and MEFs; in addition, using an in vivo study, 4T1 tumor-bearing female Balb/c mice were treated with different doses of the BCc1 nanoparticle and doxorubicin alone and together and then their mean and median survival was evaluated. Result: The results showed that the BCc1 nanoparticle increased gene expressions of RB, p53, Caspase7, p21, and Bax and decreased gene expressions of Bcl2 in MCF-7 significantly, but no change was observed in MEFs expressions. The findings revealed that the BCc1 nanoparticle, when used orally, had the highest mean and median survival time. A mixture of a high dose of the BCc1 nanoparticle (1 mg/kg) and a low dose of doxorubicin (0.1 mg/kg) showed synergistic effects on enhanced life span, while doxorubicin dose was prescribed approximately 50 times less than the murine applicable dose (5 mg/kg). Conclusion: Our results demonstrated that the BCc1 nanoparticle not only has the potential to become a novel nanomedicine for cancer therapy, but it can also provide the basis of a new medicine for cancer management when mixed with a lower applicable dose of doxorubicin.