International Journal of Reproductive Biomedicine最新文献

Background: Despite the extensive use of the gonadotropin-releasing hormone (GnRH) antagonist protocol in treating infertile women, particularly those with polycystic ovary syndrome (PCOS), there have not been sufficient evidence to compare the flexible and fixed variants in in vitro fertilization (IVF) cycles.

Objective: This study aims to assess the treatment outcomes of flexible and fixed types of GnRH-antagonist protocol for IVF in women with PCOS.

Materials and methods: In this randomized clinical trial, 150 infertile women with PCOS, who were candidates for IVF, and referred to the Yazd Research and Clinical Center for Infertility, Yazd, Iran between October 2023 and February 2024 were included. Participants were divided into 2 groups (n = 75/each) based on the type of antagonist protocol (fixed or flexible). GnRH antagonist administration started on the 5 ${\hspace{0.5em}}^{\text{th}}$ day of gonadotropin treatment in the fixed group. In the flexible group when there was at least one follicle 12-14 mm, GnRH antagonist was started. Finally, the number of metaphase II oocyte, the quality of embryos, the duration of the stimulation cycle, the dose of gonadotropin, the number of GnRH-antagonist, and the rate of ovarian hyperstimulation syndrome were evaluated.

Results: No statistically significant difference was observed in terms of cycle length and the total dose of gonadotropin between groups. Nevertheless, a notable distinction was observed in the total number of oocytes (17.84 vs. 15.5, p = 0.023) and mature oocytes (13.64 vs. 11.83, p = 0.019) in the flexible group compared to the fixed group.

Conclusion: In conclusion, the IVF outcomes are more favorable in women with PCOS undergoing the flexible GnRH-antagonist protocol compared to the fixed protocol.

Background: Carbon monoxide (CO), influences ovarian function, pregnancy, and placental health. Heme oxygenase (HO)-1 and its products, including CO, exhibit protective and anti-inflammatory properties.

Objective: This study investigates the protective effects of CO released by the carbon dioxide-releasing molecule (CORM)-2 against oxidative stress, functional and structural changes of the ovaries, and HO-1 expressions in female rats suffering from polycystic ovary syndrome (PCOS).

Materials and methods: In this experimental study, 24 Rattus norvegicus var. Albinus female rats (180-200 gr, 8 wk) were randomly divided into 4 groups (n = 6/each): control, CORM-2 (10 mg/kg), PCOS (induced by 4 mg/kg, intramuscular injection and a single dose of estradiol valerate), PCOS + CORM-2. Ovary histological changes were evaluated by crystal violet staining. Malondialdehyde (MDA) level and superoxide dismutase (SOD) activity of ovarian tissue were assessed using enzyme-linked immunosorbent assay. HO-1 expression was evaluated using Western blot.

Results: Corpus luteal formation significantly decreased in the PCOS group and was significantly restored with CORM-2 administration compared to the control group (p $<$ 0.05). The expression of ovarian HO-1 protein was reduced in the PCOS group compared to controls (p $<$ 0.01), and administration of CORM in PCOS rats significantly increased its expression (p $<$ 0.0001). In addition, CORM administration markedly reduced ovarian MDA levels and restored SOD activity (p $<$ 0.0001).

Conclusion: CORM-2 administration to PCOS rats created protective effects by reducing oxidative stress (reducing MDA level and restoring SOD activity) and increasing ovarian HO-1 protein.

Background: Melatonin and L-carnitine are free radical scavengers with antiapoptotic and antioxidant properties that improve oocyte development.

Objective: This study aimed to find the possible effect of combining 2 antioxidant agents of melatonin and L-carnitine on oocyte morphology, maturation, apoptosis, and expression of bone morphogenetic protein 15 (BMP-15) and growth differentiation factor 9 (GDF-9) genes in a mice model.

Materials and methods: To overstimulation, 60 female NMRI mice were injected intraperitoneally using mare serum gonadotropin. On day 2 post-injection, 70 cumulus-oocyte complexes were collected from each mouse. The collected oocytes randomly were then divided into 4 groups including, the control, melatonin, L-carnitine, and melatonin + L-carnitine groups. The morphology and maturation rate of the oocytes was evaluated using a light microscope. Apoptosis was identified by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay and the expression of BMP-15 and growth and differentiation factor GDF-9 genes was also evaluated by real-time polymerase chain reaction.

Results: Oocyte diameter significantly was increased in combination treatment of L-carnitine and melatonin compared to other groups (p $<$ 0.05). L-carnitine group showed the highest mean percentage of oocyte cytoplasmic pattern. Results of the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling indicated that the lowest apoptosis rate belonged to the melatonin + L-carnitine group. Moreover, the combination groups showed the highest number of oocytes and maturation rate. The BMP-15 and GDF-9 genes were significantly upregulated in all treatment groups compared to the control group.

Conclusion: Our results suggested a combination of melatonin + L-carnitine administration as a more effective choice for in vitro promotion of oocyte maturation.

Background: Follicle-stimulating hormone receptor (FSHR) and luteinizing hormone/choriogonadotropin receptor (LHCGR) are integral to ovarian function, facilitating follicle development and maturation through their respective hormonal interactions. The influence of receptor polymorphisms on the outcomes of freeze-all cycles remains unclear.

Objective: This study investigates the impact of FSHR N680S and LHCGR N312S polymorphisms on clinical outcomes in freeze-all cycles.

Materials and methods: Women undergoing controlled ovarian stimulation for assisted reproductive technology participated in this study. They were administered a gonadotropin-releasing hormone antagonist protocol, with recombinant follicle-stimulating hormone (rFSH) dosages adjusted according to age, body mass index, antral follicle count, and individual hormonal responses. Additionally, human menopausal gonadotropin dosages were tailored based on the LHCGR N312S genetic variant.

Results: Analysis revealed no significant differences in age, body mass index, antral follicle count, or marital status across the genotypes of FSHR N680S and LHCGR N312S. However, notable differences were observed in the rFSH dosage required daily and in total among the FSHR polymorphism genotypes. Genotypes of the LHCGR polymorphism correlated with fewer stimulation days. A significant interaction was observed between the 2 polymorphisms concerning total rFSH dosage.

Conclusion: The presence of serine in the FSHR polymorphism was associated with higher rFSH dosage requirements. Both FSHR N680S and LHCGR N312S polymorphisms significantly influenced clinical pregnancy and live birth outcomes in freeze-all cycles, underscoring the potential of a pharmacogenomic approach to optimize hormone supplementation in controlled ovarian stimulation protocols during assisted reproductive technology treatments.

Background: Opioid analgesics like morphine and methadone are widely used for managing severe pain; however, concerns over their potential misuse and adverse effects on the brain and reproductive system are significant.

Objective: We aimed to investigate their impacts on spermatogenesis and cognitive function in male Norway rats.

Materials and methods: In this experimental study, 36 male Norway rats (250-300 gr, 6 months old) were divided into 6 groups: low-dose morphine, high-dose morphine, low-dose methadone, high-dose methadone, positive control (received normal saline at 5 mg/kg), and negative control (received no treatment). Morphine and methadone were administered intraperitoneally over 30 days at doses of 3 mg/kg and 7 mg/kg, respectively. Behavioral assessments evaluated anxiety, stress, and short- and long-term memory. Sperm parameters (viability, motility, morphology), hormonal analysis (testosterone, luteinizing hormone, follicle-stimulating hormone, estradiol), and gene expressions (Tp53, CatSper1) were assessed.

Results: A significant reduction in rat weight was observed in the high-dose morphine group (p = 0.0045), while testicular weights remained unchanged. Sperm abnormalities were observed with high doses of methadone and morphine. High-dose methadone significantly reduced offspring count (p = 0.0004). Levels of follicle-stimulating hormone, luteinizing hormone, testosterone, and estradiol varied significantly across treatment groups. Gene expression was altered in response to treatments (p $<$ 0.05).

Conclusion: Prolonged exposure to methadone and morphine resulted in memory dysfunction, chronic stress, hormonal disturbances, altered gene expression, and fertility complications. These effects were more pronounced at higher doses, highlighting the importance of careful dosage management in opioid therapy.

Background: The lack of improvement in some endometriotic people's pain after surgery even while using hormone treatment may suggest an inappropriate response to routine hormonal therapies.

Objective: This study aimed to determine a cut-off point for selecting the most appropriate treatment based on the hormone receptors of endometriotic lesions.

Materials and methods: In this cross-sectional study, by reviewing the medical records of participants and testing their archive samples and phone interviews (if needed), 86 symptomatic women after endometriosis surgery who were operated into governmental hospitals, Shahid Faghihi and Hazrate Zeinab Shiraz Iran were enrolled between March 2017 and March 2019. Women were divided into 2 groups: responsiveness (n = 73 for dysmenorrhea, n = 60 for dyspareunia) to medical treatment and surgery, and unresponsiveness (n = 13, n = 7). We examined the pathological slides of 86 women to determine the amount of hormone receptors and the relationship between the type of medical treatment and the level of hormone receptors on pain relief within 1 yr after surgery.

Results: Based on the receiver operating characteristic curve, dysmenorrhea in the presence of tissue estrogen receptors $>$ 60% (p = 0.1065), and dyspareunia in the presence of tissue progesterone receptors $>$ 80% (p = 0.0001) responded well to medical treatment after surgery. In the presence of endometrioma-dysmenorrhea showed the best response to oral contraceptive pills (69.4%), while in deep infiltrative endometriosis-dyspareunia showed the best response to progesterone treatment (75%).

Conclusion: Prescribing an appropriate hormone therapy based on a specific immunohistochemistry staining pattern can improve the life quality of postoperative endometriosis individuals.

Background: Methadone is a substance widely used in the substitution treatment of opiate addiction in pregnancy. The placental transfer of methadone influences oxidative stress processes. Melatonin is a hormone with antioxidant activity.

Objective: This study aimed to evaluate the protective effects of melatonin on oxidative stress induced by the transfer of transplacental methadone in mice.

Materials and methods: In this experimental study, 36 female mice (2 months old, 20 $\pm$ 2 gr) were divided into 6 groups (n = 6/each) of control, methadone (0.3 mg/kg intraperitoneal, single dose) and melatonin (2, 4, and 6 mg/kg/day gavage) were administered 30 min before methadone, and one group received melatonin alone (0.6 mg/kg with single injection). Administration for 10 consecutive days of the pregnancy period was done. After baby mice were born, all neonatal mice were killed by beheading or sacrificing after anesthesia. The liver tissues were extracted. The samples were then sent for studying oxidative stress markers such as lipid peroxidation, glutathione, and protein carbonyl contents. Also, we have used the immunohistochemistry method for apoptotic markers such as: BAX, Bcl2, and Caspase3 for assaying apoptosis.

Results: This study has shown that methadone caused a significant decrease in glutathione concentration (p = 0.035). Also, we observed a significant increase in lipid peroxidation and protein carbonyl contents (p = 0.015, 0.025 respectively). However, melatonin treatment significantly inhibited oxidative stress markers (p = 0.025). Also, apoptosis assay has shown that melatonin could decrease BAX and Caspase 9 as apoptotic and increase Bcl2 as an antiapoptotic proteins (p = 0.015).

Conclusion: Our findings have shown that melatonin has a protective effect against oxidative stress and apoptosis induced by the placental transfer of methadone via its antioxidant effects.

Background: Natural killer (NK) cells play a critical role in the pathogenesis of endometriosis. Moreover, a normal vitamin D level is remarkably associated with an optimal immune response. So, there may be a probable relationship between these factors and the endometriotic women.

Objective: This study aimed to evaluate the percentage of NK cells and their subsets and their relationship with serum levels of vitamin D and interferon-gamma (IFN-γ) in women with endometriosis.

Materials and methods: In this case-control study, 29 women with stage III-IV endometriosis and 30 healthy controls were enrolled. The study was conducted in the Immunology Department of Isfahan University of Medical Sciences, Isfahan, Iran between November 2021 and June 2022. The percentage of NK cells and their subsets, including CD56 ${\hspace{0.5em}}^{\text{dim}}$ CD16 ${\hspace{0.5em}}^{+}$ , CD56 ${\hspace{0.5em}}^{\text{bright}}$ CD16 ${\hspace{0.5em}}^{-}$ and CD56 ${\hspace{0.5em}}^{\text{bright}}$ CD16 ${\hspace{0.5em}}^{\text{bright}}$ were measured in the peripheral blood samples using flow cytometry. Serum levels of vitamin D and IFN-γ were also measured using the enzyme-linked immunosorbent assay.

Results: The mean percentage of NK cells in women with endometriosis increased significantly compared to the control group (p = 0.03). The percentage of CD56 ${\hspace{0.5em}}^{\text{dim}}$ CD16 ${\hspace{0.5em}}^{+}$ (p = 0.007) and CD56 ${\hspace{0.5em}}^{\text{bright}}$ CD16 ${\hspace{0.5em}}^{\text{bright}}$ (p = 0.043) increased significantly in women with endometriosis in comparison with the control group, but the percentage of CD56 ${\hspace{0.5em}}^{\text{bright}}$ CD16 ${\hspace{0.5em}}^{-}$ subset was not significantly different. No relationship was observed between NK cells and their subsets with vitamin D and IFN-γ in the studied groups.

Conclusion: The study of NK cell subsets and their related factors can be useful in assessing and treating women suffering from endometriosis. However, more comprehensive studies are required to draw definitive conclusions about these observations.