Investigating the impact of local inflammation on granulosa cells and follicular development in women with ovarian endometriosis

Objective: To investigate the possible impact of local inflammation on granulosa cells (GCs) and follicular development in endometriosis patients. Design: Prospective study with related paired design. Setting: Reproductive medicine center. Patient(s): A total of 80 endometriosis patients and 104 controls, with cultured GCs collected from control participants younger than 35 years. Intervention(s): Tumor necrosis factor-alpha (TNF-alpha) and nuclear factor kappa B (NF-kappa B) inhibitor. Main Outcome Measure(s): Intrafollicular concentrations of cytokines measured with ELISA, NF-kappa B binding levels with electrophoretic mobility shift assay (EMSA), and telomerase activity (TA) with quantitative-telomeric repeat amplification protocol (Q-TRAP) assay, and protein and mRNA expression with Western blot and polymerase chain reaction analyses, respectively. Result(s): Patients with endometriosis exhibited a statistically significantly lower antral follicle count (11.48 +/- 8.11 vs. 15.68 +/- 8.56), lower number of retrieved oocytes (8.28 +/- 6.69 vs. 10.87 +/- 6.26), and lower number of mature oocytes (6.67 +/- 6.09 vs. 8.53 +/- 5.69). The GCs from endometriosis patients showed higher NF-kappa B binding activity and increased expression of inhibitor of NF-kappa B kinase subunit beta (IKK beta, 2.743-fold) and NF-kappa B inhibitor alpha (I kappa B alpha, 5.017-fold). Their NF-kappa B p65 expression was negatively associated with mature oocytes (bNF-kappa B' = -0.304, R-2 = 0.195, R = 0.442) but positively associated with intrafollicular TNF-alpha (r = 0.37); TA showed a negative relationship with NF-kappa B binding levels (r = -0.667). Tumor necrosis factor-alpha induced expression of I kappa B alpha (5.408-fold) and NF-kappa B p65 (1.400-fold) but lowered human telomerase reverse transcriptase (hTERT) and TA levels (0.0009 vs. 0.5619) in cultured GCs. However, inhibiting NF-kappa B obviously increased hTERT expression (1.988-fold). Conclusion(s): Endometriosis showed activated NF-kappa B pathways in GCs, which might negatively affect TA and oocyte quality. Intrafollicular TNF-alpha might down-regulate TA and hTERT via NF-kappa B pathway, but further studies are required. ((C) 2019 by American Society for Reproductive Medicine.)