F&S science最新文献

Objective: Stearoyl-CoA desaturase (SCD1) is an enzyme that catalyzes the conversion of saturated delta-9 fatty acids to monounsaturated fatty acids. SCD1 is highly expressed in various cancers and facilitates cancer cell survival, tumor growth, and metastasis. This study aimed to assess SCD1 expression and function in uterine leiomyoma and matched myometrial tissue and evaluate the impact of SCD1 inhibition on leiomyoma cell viability and apoptosis.

Design: Gene set enrichment analysis was performed to determine whether lipid metabolism pathways are dysregulated in leiomyoma. To assess the function of SCD1, primary leiomyoma and myometrial cells, as well as a CRISPR-engineered leiomyoma-relevant MED12 mutant human uterine smooth muscle (UtSM) cell line, were treated with SCD1 small interfering RNA or a small molecule inhibitor of SCD1, CAY10566. Cell viability and apoptosis assays, real-time quantitative polymerase chain reaction, and immunoblot analyses were performed to evaluate cell function in response to treatment.

Subjects: Leiomyoma and myometrial tissues were obtained from premenopausal individuals designated female at birth (n = 30) undergoing myomectomy or hysterectomy.

Exposures: SCD1 inhibition by small interfering RNA and CAY10566 treatment.

Main outcome measures: Messenger RNA (mRNA) and protein levels and cell viability and apoptosis.

Results: Gene set enrichment analysis revealed that the cholesterol homeostasis pathway was significantly different in leiomyoma vs. adjacent myometrial tissues. Among the genes in this pathway, SCD1 mRNA levels were found to be significantly higher in leiomyoma than in matched myometrium. SCD1 inhibition by small interfering RNA or CAY10566 decreased antiapoptotic BCL2 mRNA and protein levels and cell viability in primary leiomyoma but not myometrial cells. SCD1 protein levels were significantly higher in the mutant MED12 UtSM cell line than in the wild-type MED12 UtSM cell line. CAY10566 treatment specifically decreased cell viability and increased apoptosis in mutant MED12 UtSM cells, with increased protein levels of cleaved caspase 3, cleaved PARP, and DDIT3 in mutant MED12 UtSM but not in wild-type MED12 UtSM cells.

Conclusion: SCD1, an enzyme involved in lipid homeostasis, may play an important role in promoting leiomyoma growth and represents a novel target for the treatment of leiomyoma.

Objective: To determine the effect of an obesogenic Western-style diet and hyperandrogenemia on ovarian outcomes.

Design: Experimental, controlled animal study SUBJECTS: Post-pubertal rhesus macaque females EXPOSURE: A Western-style diet (T+WSD: 36% fat, 45% carbohydrate, 18% protein) combined with exogenously administered testosterone versus a standard chow diet (Control, CTRL; 15% fat, 59% carbohydrate, 27% protein). Animals underwent controlled ovarian stimulations to assess ovarian follicle development.

Main outcome measures: Cycle length, the proportion of ovulatory cycles, and daily levels of estradiol (E2), progesterone (P4), AMH, luteinizing hormone (LH), and follicle-stimulating hormone (FSH) were compared between CTRL and T+WSD groups through one menstrual cycle. Follicular fluid was assessed for cytokine and steroid content, and retrieved oocytes were evaluated for meiotic maturation and underwent in vitro fertilization. Granulosa cells were analyzed for differential gene expression. Ovaries were removed in early luteal phase (4 days post midcycle estradiol surge) and analyzed for morphological differences.

Results: The T+WSD group demonstrated significantly decreased luteal P4 levels. We found no differences in cycle length, proportion of ovulatory cycles, day of E2 surge, total E2 synthesis, FSH, LH, or AMH. Analysis of follicular fluid retrieved from animals undergoing an ovarian stimulation protocol revealed increased vascular endothelial growth factor-A (VEGFA), elevated cortisol:cortisone ratio, and increased testosterone and progesterone levels in the treatment group (p<0.05). Granulosa cells from T+WSD demonstrated significantly up- or down-regulated genes relative to controls, including those related to cell differentiation and migration. The ovarian morphology of treatment animals demonstrated enlarged cystic follicles reminiscent of polycystic ovaries.

Conclusion: Similar to prior studies assessing long-term exposure (5-6 years) to T+WSD in female rhesus macaques beginning before menarche, a 1-year T+WSD treatment in adult, regularly cycling females led to reduced luteal-phase progesterone levels and polycystic ovarian morphology. Additionally, short-term T+WSD exposure resulted in altered granulosa cell gene expression. While 1 year of T+WSD exposure leads to altered luteal progesterone, follicular fluid steroid and cytokine content, and granulosa cell gene expression changes, insults of longer duration are required to exert additional negative effects on ovarian function.

Objective: To study the effects of generally considered safe doses of antioxidant micronutrient supplementation on semen parameters, systemic redox balance, sperm DNA structural integrity, and fertility.

Design: Given ethical limitations in human studies, this dose escalation study examined the effects of common water-soluble antioxidant micronutrients (Vitamin C, Zinc, Folate, and Carnitine) on semen parameters, redox status, DNA integrity, and fertility outcomes in healthy male mice over one spermatogenic cycle. The study was partially repeated at the highest carnitine dose for pregnancy outcomes and comparatively assessed in subfertile, oxidatively stressed mice.

Subjects: "Fertile/healthy" (CD1) and "Subfertile/oxidatively stressed" (gpx5^-/-) mice.

Exposure: Water-soluble micronutrients (Vitamin C, Zinc, Folate, and Carnitine).

Intervention: N/A.

Main outcome measures: Sperm parameters included count, motility, viability, and acrosome integrity. Systemic redox status was evaluated in blood, measuring malondialdehyde, thiol levels and total antioxidant capacity. Sperm DNA parameters were examined for oxidation (8-OHdG staining), fragmentation (TUNEL), and decondensation (Toluidine Blue). Pregnancy outcomes were also assessed in CD1 mice fed carnitine.

Results: In healthy mice, increasing doses of individual micronutrients had minimal effects on semen parameters. However, high doses of all four micronutrients significantly disrupted the redox balance in blood plasma and compromised sperm DNA integrity in an ingredient-specific manner. Moderate to high doses of carnitine caused severe DNA fragmentation, a finding confirmed in a subsequent experiment using the highest carnitine dose. In this follow-up experiment, male mice supplemented with carnitine and mated with females showed decreased pregnancy rates and fewer total pups born. Conversely, in oxidatively stressed mice, high-dose carnitine had the opposite, beneficial effect of improving sperm DNA integrity.

Conclusion: At high doses, antioxidants can induce reductive stress, damaging vital molecular components of sperm cells such as DNA. While strong evidence supports the use of preconception antioxidants to boost semen quality, healthcare professionals should assess oxidative stress levels when possible and recommend personalized antioxidant doses to avoid reductive stress and prevent adverse reproductive outcomes.

Objective: To assess the effects of the active metabolite of Bupropion (BUP), hydroxybupropion (OH-BUP), on human spermatozoa in vitro.

Design: Laboratory based in vitro study.

Subjects: Human sperm samples were collected from 45 normozoospermic patients (smokers and non-smokers).

Exposure: Sperm samples were incubated at 37°C with 5% CO₂ for two hours with and without the IC₅₀ concentration of OH-BUP (1.9 μM).

Main outcome measures: Several sperm analysis were performed, including vitality, motility, chromatin condensation, DNA fragmentation, oxidative stress, mitochondria membrane potential, and acrosome integrity. High-speed video microscopy and Computer-Assisted Sperm Analysis provided a detailed evaluation of sperm motility.

Results: OH-BUP exposure resulted in significant impairments in sperm vitality and motility, particularly in progressive motility. Chromatin condensation was significantly reduced, while DNA fragmentation, especially in the equatorial region, significantly increased. Mitochondria membrane potential, acrosomal integrity, and reactive oxygen species production also significantly decreased following exposure.

Conclusion: The findings indicate potential harm to sperm parameters, which may contribute to infertility. Understanding how BUP affects reproductive function is crucial for clinicians and patients to make informed decisions regarding the use of this medication.

Objective: To clearly elaborate the causal relationships between specific immunocyte phenotypes and male infertility.

Design: Mendelian randomization using genome-wide association study data.

Setting: Publicly available genome-wide association study data.

Subjects: Large cohorts of European ancestry.

Exposure: 731 immunocyte phenotypes or male infertility.

Main outcomes measures: Genetic variants were used as instrumental variables to infer causality, minimizing confounding and bias. The causal associations were assessed using the inverse variance-weighted (IVW) method for primary analysis and validated the findings using MR-Egger, Weighted Median, Simple Mode, and Weighted Mode approaches. Additional sensitivity analyses were performed to validate the robustness of findings.

Results: Our analysis identified significant causal associations between specific immunocyte phenotypes and male infertility. Phenotypes such as Naive-mature B cell %lymphocyte (OR = 1.257, P = 0.009) and IgD- CD38dim %B cell (OR = 1.100, P = 0.021) were positively associated with increased infertility risk, while phenotypes like CD39+ CD8br %T cell (OR = 0.856, P = 0.021) and BAFF-R on transitional (OR = 0.833, P = 0.002) were negatively associated, suggesting a protective effect. Additionally, reverse MR analysis revealed that male infertility might causally affect certain immunocyte phenotypes, including CD14- CD16+ monocyte %monocyte (OR = 1.049, P = 0.007).

Conclusions: This study provides robust evidence for the causal role of specific immunocyte phenotypes in male infertility and highlights the bidirectional relationship between immune function and reproductive health. These findings provide new insights into the immunological factors contributing to male infertility and suggest potential biomarkers and therapeutic targets for future research and clinical interventions.

Objective: To identify key genes and potential drug targets for ovarian-related diseases through genome-wide Mendelian randomization (MR) and colocalization analyses.

Design: We conducted a comprehensive two-sample MR analysis to estimate the causal effects of blood expression quantitative trait loci (eQTLs) on ovarian-related diseases, followed by colocalization analyses to verify the robustness of the expression instrumental variables (IVs). Phenome-wide association studies (PheWAS) were also performed to evaluate the horizontal pleiotropy of potential drug targets and possible side effects.

Setting: Publicly available genome-wide association study data.

Subjects: Large cohorts of European ancestry.

Exposure: The exposure in this study was the genetic variants (eQTLs) associated with gene expression levels, considered a form of lifelong exposure. eQTL data were obtained from the eQTLGen Consortium, encompassing 16,987 genes and 31,684 cis-eQTLs derived from blood samples of healthy individuals of European ancestry.

Main outcome measure: The primary outcome measures were the identification of genes causally associated with ovarian-related diseases and the validation of these genes as potential therapeutic targets.

Results: Our study revealed that specific genes such as CD163L1, PPP3CA, MTAP, F12, NRM, BANK1, ZNF66, GNA15, and SLC6A9 were associated with ovarian endometriosis, ovarian cysts, and PCOS. Through MR and colocalization analyses, we identified potential drug targets, including CTNNB1, PTPN7, and ABCB4, with strong evidence of colocalization with ovarian-related diseases. Sensitivity analyses confirmed the robustness of our findings, showing no evidence of horizontal pleiotropy or heterogeneity.

Conclusion: This research highlights the significance of precision medicine approaches in identifying genetic factors underlying ovarian-related diseases and provides a foundation for developing targeted therapies, enhancing diagnostic accuracy, and improving treatment strategies for ovarian-related diseases.

Objective: To determine if microRNAs that are altered in the circulation of women with endometriosis affect metabolic gene expression in hepatic cells.

Design: In vitro study.

Setting: Academic Medical Center. HUMAN HEPG2 CELLS: MiR-let-7b, miR-125b-5p, miR-150-5p, and miR-3613-5p, their inhibitors or respective controls were transfected in human HepG2 cells.

Intervention(s): MicroRNAs were used to induce or suppress target genes in hepatic cells.

Main outcome measure(s): Effect of the microRNAs that are aberrantly expressed in endometriosis on hepatic cell gene expression using qPCR.

Result(s): Prior microarray studies on serum of women with endometriosis showed differential expression of microRNAs miR-Let-7b, miR-125b-5p, miR-150-5p, and miR-3613-5p. Bioinformatic analyses revealed that these microRNAs have predicted binding sites in multiple genes that are involved in liver metabolism. Transfection of these miRs in HepG2 cells followed by qRT-PCR showed that miR-Let-7b mimic increased the expression of Igfbp1 by 8-fold (p = 0.004) and reduced the expression of Mrc1 by 3.2-fold (p = 0.003) while its inhibitor reduced Igfbp1 by 2.8-fold (p = 0.0004) and increased Mrc1 by 5.2-fold (p = 0.04). MiR-3613-5p mimic reduced the expression of Cyp2r1 by 2.2-fold (p = 0.04) and Mrc1 by 4-fold (p= 0.0001). MiR-125b-5p mimic increased the expression of Fabp4 by 4.1-fold (p = 0.020) while miR-150-5p mimic increased the expression of Mrc1 by 1.8-fold (p = 0.01) and Cyp2r1 by 2.5-fold (p = 0.008). Inhibitors of both miR-125b-5p and miR-150-5p did not show any effect on any of the genes.

Conclusion(s): Circulating microRNAs known to be aberrant in endometriosis regulated hepatic gene expression, likely contributing to the metabolic defects seen in this disease. Treatment with miR-Let-7b and miR-3613-5p, which are downregulated in endometriosis, reversed the effect of endometriosis on the expression of IGFBP1, MRC1 and CYP2r1 genes. Therefore, miR-Let-7b and miR-3613-5p may be novel candidate therapies for endometriosis, potentially correcting the metabolic changes seen in endometriosis patients.