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Introduction: Teratozoospermia is a common cause of human infertility. The percent of live sperm in each specimen is a critical factor for determination of fertility rate. It has been shown that sustained intracellular calcium overload plays a major role in apoptosis. We investigated the biological significance of apoptosis in normozoospermia relative to teratozoospermia in the presence of calcium ionophore. Materials and Methods: Teratozoospermic (n=8) and normozoospermic (n=10) semen samples were selected based on WHO criteria. Washed sperm incubated in sperm medium, DMSO, or calcium ionophore A23187 (10 μM) at 37o° for 1 h under 5% CO2. Apoptosis in all experimental groups were analyzed by flow cytometry using the Annexin V/propidium iodide (PI) binding assay. Results: Control and DMSO groups of teratozoospermia had significant higher level of necrotic and dead cells compared to normozoospermia; however, the percent of early and late apoptosis did not show any significant differences. The percent of early and late apoptosis increased in normozoospermia and teratozoospermia after incubation with calcium ionophore. No significant differences were found in the level of necrotic cells between control and ionophore treated groups. Conclusion: Our results suggest that human sperm are susceptible to apoptosis in the presence of sustained intracellular calcium overload; however, the percent of necrotic cells did not change. This revealed that apoptosis and necrosis in sperm have different inducers. [ABSTRACT FROM AUTHOR]

Polycystic ovary syndrome (PCOS) is accompanied with many disturbances in hormone synthesis and antioxidant defense. Previous reports have indicated that Vitamin D (vit.D) affects gene expression and have roles in normal follicular development. Therefore, we investigated the effects of vit.D on steroidogenesis, apoptosis, reactive oxygen species (ROS) production, and antioxidant defenses of human normal granulosa cells (N-GCs) and granulosa cells from polycystic ovaries (PCO-GCs). Ovarian GCs were obtained during oocyte retrieval procedure from 120 women with PCOS and from 100 healthy women who referred to Shiraz Fertility Center. The isolated GCs were cultured in the presence or absence of vit.D (100 nM), for 48 h. Concentration of sex steroids was measured by ELISA. Glutathione peroxidase (GPx) and superoxide dismutase (SOD) expression and activities were assessed by q-PCR and photometric methods, respectively. The amount of ROS production was estimated using chemiluminescence and fluorescence methods. Cell viability and apoptosis were detected by Annexin-V/propidium iodide detection kit. Basal estrone and progesterone secretion by N-GCs was significantly higher than that of PCO-GCs. Vit.D significantly increased aromatase and 3β-hydroxysteroid dehydrogenase activity in N-GCs and PCO-GCs. Basal expression and activity of GPx, in PCO-GCs were significantly lower than those of N-GCs. Treatment with vit.D significantly increased genes expression and enzyme activities in both groups. Basal ROS in PCO-GCs was markedly greater than that of N-GCs, which was attenuated by vit.D treatment. Cell apoptosis was directly correlated with ROS levels. We conclude that vit.D improved N-GCs and PCO-GCs functions through affecting steroidogenesis and enzymatic antioxidant defense. Under vit.D treatment, PCO-GCs could act more similar to N-GCs., Competing Interests: Declaration of Competing Interest The authors declare no competing financial interest., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Calcium (Ca(2+)) and reactive oxygen species (ROS) constitute the most influential intracellular signaling molecules participating in the regulation of different sperm functions. Elevating intracellular Ca(2+) and ROS in physiologic range regulate capacitation, motility, acrosome reaction, and sperm-oocyte fusion; whereas cytosolic Ca(2+) overload and ROS overproduction have pathologic effects. Our aim of this study was determination of antioxidant effects of α-tocopherol on sperm motility, viability, and DNA integrity in a condition where cytosolic calcium overload was induced by A23187 (a calcium ionophore). Our results indicated that, α-tocopherol has ability to prevent sperm mortality and save sperm rapid motility after 1 hour incubation. At the same time, A23187 reduced significantly percentage of rapid sperm motility and increased sperm mortality and DNA damage. Results of sperms incubation in the medium contain a combination of A23187 and α-tocopherol showed that α-tocopherol can reduce many of the deleterious effects of A23187. In conclusion, it seems that the harmful effects of A23187 are due to excessive ROS production, and α-tocopherol neutralizes these effects.