Bilateral varicocele leads to ferroptosis, pyroptosis and necroptosis of human spermatozoa and affects semen quality in infertile men.

Purpose: This study explored the effects of bilateral varicocele on male semen quality in infertile men and the molecular mechanisms involving ferroptosis, pyroptosis and necroptosis signaling pathways. Methods: Totally, 20 healthy males and 26 patients with bilateral varicocele receiving infertility treatment were enrolled. Semen samples were collected. Basic semen parameters, acrosome integrity and membrane integrity, mitochondrial membrane potential (MMP) and apoptosis rate were compared. Levels of reactive oxygen species (ROS), iron, glutathione (GSH), total superoxide dismutase (T-SOD), and, Catalase (CAT), were detected in human seminal plasma. Relative mRNA expression of Ca 2+-independent phospholipases A2 beta (iPLA 2ß), P53, Zinc finger E-box binding homeobox 1 (ZEB1) and GSH-dependent peroxidase 4 (GPX4) were evaluated. Relative protein expression was determined for GPX4, receptor interacting serine/threonine kinase 1 (RIPK1) and receptor interacting serine/threonine kinase 3 (RIPK3), as well as pyroptosis markers of Gasdermin E (GSDME) and heat shock protein 90 (HSP 90). Results: The results revealed that the bilateral varicocele group had significantly higher abnormalities (sperm progressive rate and sperm motility) compared to the control group. Meanwhile, compared to control group, GSH, T-SOD, and CAT levels were reduced in the bilateral varicocele group (p < 0.05). However, the level of ROS and iron were significantly increased (p < 0.05). Relative mRNA expression of P53, iPLA 2ß, ZEB1, and GPX4 were reduced. In addition, ROS exposure activated ferroptosis-related signal pathways. RIPK1, RIPK3, GSDME and HSP 90 were increased in bilateral varicocele group. ROS exposure affected signaling pathways related to ferroptosis, necrosis and pyroptosis in human spermatozoa. Conclusion: Bilateral varicocele leads to ferroptosis, pyroptosis and necroptosis of human spermatozoa and affects semen quality in infertile men.

The status and comparison of ovarian reserve between fertile and infertile healthy Chinese women of reproductive age.

ABSTRACT: We aimed to investigate ovarian reserve status, and explore differences in ovarian reserve between fertile and infertile healthy Chinese women of reproductive age.We recruited 442 fertile women aged 23 to 49 years (mean: 35.22 ±â€Š4.91 years) as subjects, and 196 infertile women aged 23 to 46 years (mean: 32.34 ±â€Š4.34 years) as controls. For all participants, a number of parameters were tested on days 2 to 4 of a spontaneous cycle, including basal serum follicle-stimulating hormone (FSH), estradiol (E2), luteinizing hormone (LH), total testosterone, anti-Müllerian hormone (AMH), ovarian response prediction index (ORPI), and antral follicle count (AFC).There were significant differences in terms of AFC, serum AMH levels, and ORPI among subject subgroups (10.58 ±â€Š5.80; 2.533 ±â€Š2.146 ng/mL; 1.28 ±â€Š1.87; respectively), and among control subgroups (12.44 ±â€Š5.69; 3.189 ±â€Š2.551 ng/mL; 1.88 ±â€Š2.68; respectively) (P < .01 for all). For both subjects and controls, AFC, AMH levels, and ORPI decreased gradually with increasing age, and presented with similar age-related trends; there were positive correlations between AMH and AFC (P < .001), and negative correlations between age and AFC, AMH, ORPI (P < .05 for all). There was a significant difference in age (P < .001), serum E2 (P < .01), and AMH (P < .01) levels between subjects and controls; however, when controlling for confounding factors (age, body mass index, total testosterone, and LH), we found no differences between the 2 groups with regards to the serum levels of AMH, FSH, E2, and AFC (P > .05 for all). Moreover, receiver operating characteristic curve analysis indicated that the significant variables of subjects and controls for evaluating ovarian reserve included age, AMH and ORPI, and ORPI was more valuable than other variables.A diminished ovarian reserve was one of the manifestations caused by female aging. When confounding factors were controlled for, we found no differences in ovarian reserve when compared between fertile and infertile women, and no correlation with infertility.

High anti-Müllerian hormone levels might not reflect the likelihood of clinical pregnancy rate in IVF/ICSI treatment.

OBJECTIVE: To investigate if high anti-Müllerian hormone (AMH) concentration is a useful tool to predict the outcome of assisted reproductive treatment. METHODS: Retrospective cohort study involving 520 patients who underwent IVF/ICSI procedures in a university hospital. We measured the serum AMH level on day 3 of the menstrual cycle. Based on AMH levels, we divided the patients into three groups as follows: low (<25th percentile) AMH group, average (25th to 75th percentile) AMH group and high (>75th percentile) AMH group. We recorded the fertilization rate (FR), the number of oocytes retrieved, the number of good quality embryos (GQEs) and the clinical pregnancy rate (CPR). RESULTS: There was no difference between the three AMH groups in terms of maternal age, body mass index (BMI), follicle-stimulating hormone (FSH), estradiol (E2), luteinizing hormone (LH) and testosterone (T) in the IVF/ICSI cycles. The women in the high serum AMH group had a higher number of retrieved oocytes than those in the low or average AMH groups (p < 0.01) in the IVF/ICSI cycles. Compared with the low or average AMH groups, the women with high AMH levels had a higher number of good quality embryos (GQEs) in the IVF/ICSI cycles (p < 0.01). However, high AMH women had no significantly higher clinical pregnancy rate (CPR) compared to the women in the low or average AMH groups. In addition, for the prediction of CPR, the AMH levels alone were not an independent predictor of CPR for IVF and ICSI cycles in the ROC curve analysis. CONCLUSIONS: High anti-Müllerian hormone levels are an independent predictor of the number of retrieved oocytes and good quality embryos (GQEs), but might not reflect the likelihood of higher clinical pregnancy rates (CPR) in IVF/ICSI treatment.

Melatonin Inhibits Oxidative Stress and Apoptosis in Cryopreserved Ovarian Tissues via Nrf2/HO-1 Signaling Pathway.

In the field of assisted reproductive technology, female fertility preservation, particularly ovarian tissue cryopreservation in adolescent cancer patients, has attracted much attention. Melatonin (MLT) is well known for its antioxidative and anti-apoptotic properties; however, whether it can ameliorate the cryoinjury and inhibit the generation of reactive oxygen species (ROS) in cryopreserved ovarian tissues (OTs) has not yet been reported. Here, we demonstrated that MLT could protect follicular integrity; prevent cell apoptosis; decrease ROS, malondialdehyde (MDA), and nitric oxide (NO) levels; and increase activities of glutathione peroxidases (GSH-Px), glutathione (GSH), catalase (CAT), and superoxide dismutase (SOD) in cryopreserved OTs. Furthermore, these effects may be related with the activation of the nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway, as evidenced by increased mRNA levels of Nrf2 downstream genes, including heme oxygenase-1 (HO-1), glutathione S-transferase M1 (GSTM1), SOD, and CAT. In summary, MLT can not only directly scavenge ROS but also significantly induce the activation of antioxidative enzymes via the Nrf2 signaling pathway, which is a new mechanism underlying the protection effects of MLT on cryopreserved OTs.