F&S science最新文献

Objective: To determine if engineered high-mobility group AT-hook 2 (HMGA2) overexpressing uterine primary myometrial cells recapitulate the transcriptional and epigenomic features of HMGA2-subtype leiomyomas.

Design: Isolated primary, "normal" myometrial cells from three patients were engineered to overexpress HMGA2 to determine how HMGA2 establishes transcriptomic and epigenomic features of HMGA2-overexpressing leiomyoma.

Setting: Academic research laboratory.

Patient(s): Primary myometrial cells were isolated from normal myometrium obtained from three patients undergoing hysterectomy.

Intervention(s): Not applicable.

Main outcome measure(s): Determined genome-wide transcriptomic and epigenomic features of engineered HMGA2-overexpressing uterine primary myometrial cells.

Result(s): Engineered HMGA2-V5-overexpressing primary myometrial cells approximated the HMGA2 expression level observed in HMGA2-overexpression subtype leiomyoma. High-mobility group AT-hook 2-V5 expression resulted in differential expression of 1,612 genes (false discovery rate [FDR] < 0.05) that were found to be enriched in pathways associated with leiomyoma formation, including extracellular matrix organization. Comparative gene expression analysis between HMGA2-V5 engineered primary cells and HMGA2-overexpression subtype leiomyoma revealed significant overlap of differentially expressed genes. Mechanistically, HMGA2-V5 overexpression resulted in 41,323 regions with differential H3K27ac deposition (FDR < 0.05) and 205,605 regions of altered chromatin accessibility (FDR < 0.05). Transcription factor binding site analysis implicated the AP-1 family of transcription factors.

Conclusion(s): High-mobility group AT-hook 2 overexpression induces leiomyoma-like transcriptomic and epigenomic modulations in myometrial cells.

Objective: To study the role of PGE₂ in regulating plasminogen activator inhibitor-1 (PAI-1) and tissue plasminogen activator (tPA) in human primary endometrial endothelial cells (HEECs) from women with normal menstrual bleeding (NMB) and heavy menstrual bleeding (HMB).

Design: In vitro study using endometrial endothelial cells.

Setting: Research laboratory setting.

Patients: Women with NMB and HMB provided endometrial biopsy samples.

Interventions: Prostaglandin E₂ and PGE₂ receptor-selective agonists were administered to cultured HEECs.

Main outcome measures: Levels of PAI-1 and tPA in NMB-HEECs and HMB-HEECs after treatment with PGE₂ and receptor-selective agonists.

Results: Prostaglandin E₂ increased total PAI-1 levels in NMB-HEECs, but not in HMB-HEECs, which had higher baseline PAI-1 levels. PGE₂ receptors (PTGER)1 and PTGER2 agonists increased PAI-1 in NMB-HEECs, whereas PTGER3 and PTGER4 did not. Prostaglandin E₂ had no effect on tPA levels in either NMB-HEECs or HMB-HEECs.

Conclusions: Prostaglandin E₂, through PTGER1 and PTGER2, regulates the plasminogen activator system in NMB-HEECs, suggesting a role in reducing fibrinolytic activity during normal menstrual cycles. The lack of PGE₂ effect and elevated baseline PAI-1 in HMB-HEECs support using this in vitro model to further understand prostaglandin pathways in NMB and HMB.

Objective: To examine the effect of skoochies, an illicit cocktail drink, on testicular and sperm function in male rats.

Design: Twenty-five adult male Wistar rats were assigned randomly into five groups (n = 5) as follows: normal saline; skoochies; Cannabis sativa; codeine; and tramadol. The cocktail (skoochies) used in this study was formulated with the following composition: codeine (5 mg/kg); tramadol (20 mg/kg); and cannabis extract (2 mg/kg). These doses are as previously reported. Administration was performed once daily for 28 days.

Setting: University.

Animal(s): Twenty-five (25) male Wistar rats.

Intervention(s): Skoochies, tramadol, Codeiene, Cannabis.

Main outcome measure(s): Skoochies and its components induced testicular and sperm damage via increased generation of reactive oxygen species and impairment of glutathione system in rats.

Result(s): Skoochies increased reactive oxygen species generation and impaired the antioxidant system resulting in inflammation that eventually damaged the testicular tissue. Skoochies caused oxidoinflammatory injury to this tissue, leading to impaired testicular function. This was evident by the distorted cytoarchitecture, reduced sperm count and motility, and impaired testicular deoxyribonucleic acid integrity.

Conclusion(s): Thus, our results infer that skoochies impaired the testicular and sperm function through the increased generation of reactive oxygen species and impairment of the glutathione system.

Objective: To evaluate the impact of coenzyme Q-10 (CoQ-10) on the dysregulated synthesis of extracellular matrix proteins mediated by transforming growth factor beta 3 (TGF-β3) in uterine leiomyomas.

Design: Laboratory study.

Setting: University.

Patients: None.

Interventions: Treatment of immortalized uterine myometrial and leiomyoma cells to TGF-β3 and CoQ-10.

Main outcome measures: The protein concentrations of collagen 1A1 (COL1A1), collagen 3A1 (COL3A1), collagen 11A1 (COL11A1), and fibronectin (FN1) were assessed through western blot analysis after treatment of immortalized uterine myometrial and leiomyoma cells with both transforming growth factor beta (TGF-β) 3 and concentrations of CoQ-10 at 10, 50, and 100 μM concurrently for 24 hours.

Results: Immortalized uterine leiomyoma and myometrial cells exposed to TGF-β3 for 24 hours demonstrated a significant up-regulation of COL1A1, COL3A1, COL11A1, and FN1 compared with untreated cells. In leiomyoma cells, concurrent treatment with CoQ-10 over the same timeframe revealed a dose-dependent decrease in these protein concentrations compared with those in cells treated with TGF-β3 alone. At the highest concentration of 100 μM of CoQ-10, significant decreases in the amounts of COL1A1 (0.59 ± 0.10-fold), COL3A1 (0.46 ± 0.09-fold), COL11A1 (0.53 ± 0.09-fold), and FN1 (0.56 ± 0.09-fold) were observed. Similarly, myometrial cells exposed to both TGF-β3 and CoQ-10 demonstrated a dose-responsive decline in the amount of extracellular matrix protein compared with cells exposed to TGF-β3 alone. Significant reductions in the amounts of COL1A1 (0.75 ± 0.03-fold), COL3A1 (0.48 ± 0.06-fold), COL11A1 (0.38 ± 0.06), and FN1 (0.69 ± 0.04-fold) were appreciated at 100-μM CoQ-10.

Conclusion: Coenzyme Q-10 mitigated the aberrant production of key biomarkers of the extracellular matrix mediated by TGF-β3 in uterine leiomyomas. Our findings highlight a promising nonhormonal compound that can counteract the fibroproliferative process inherent to leiomyomas.