Response of endometrial semaphorins in different COH treatment protocols: In vitro study

OBJECTIVE: Semaphorins have been identified as axonal guidance molecules and later found in non-neuronal tissues. The semaphorin family has many potential functions including angiogenesis, adhesion and invasive roles, and a guidance mechanism during epithelial-stromal communication and implantation. Adhesion, invasion and angiogenesis during implantation are major physiological changes, and semaphorin 3 family homologs have been shown to decrease in the implantation window. It is thought that SEMA reduction in endometrial cells may be beneficial in implantation-based infertility. For this purpose, we investigated in vitro conditions that letrozole (L), gonadotropin (Gn) and growth hormone (GH) drugs used in ovulation induction in our study altered class 3 semaphorins secreted in the endometrium. MATERIALS AND METHODS: In vitro drug efficacy was determined using the CRL-1671 human endometrial epithel cell line. A total of 6 groups were formed, the control group being the first group. Group 2 GH, Group 3 Gn, Group 4 GH + Gn, Group 5 L + Gn, Group 6 L + Gn + GH. Drug combinations were applied to the clinical applications in accordance with appropriate protocols on days 1, 4 and 8 on GH, 5 days on L, 8 days on Gn (reduced by 1/3 when applied with L and on day 3). Immunohistochemistry and immunofluorescence staining were performed and the samples were evaluated at 200x magnification. Staining intensity (i) of positive stained cells in every 100 cells in 5 randomly selected 5 sites for each preparation; HSCORE values were determined as being mild (1) (+), moderate (2) (++), and severe (3) (+++), [HSCORE: ∑Pi (i + 1) (Pi:% of cells stained positive; i: staining intensity)]. The results were evaluated by One-way ANOVA statistical test. Statistically, p values less than 0.05 were considered statistically significant (* p<0.05, ** p<0.01, *** p<0.001). RESULTS: The decrease in SEMA 3A, 3F and 3E in the GH and GH groups (GH + Gn and GH + Gn + L groups), in the CRL-1671 human endometrial epithel cell line was statistically significant when compared to the control group (p <0.001). When the group GH is compared with the group GH + Gn groups, no statistically significant difference was found (p> 0.05). When the group GH + Gn is compared with the group GH + Gn + L; there was no statistically significant difference (p> 0.05). CONCLUSION: Growth hormone has been shown to reduce semaphorin 3A, 3E and 3F in the human endometrial epithel cell line. Letrozole and gonadotropins have been shown to not contribute to this effect. It has also been shown that the growth hormone letrozole joint has no superiority at this point and that it is better to choose growth hormone instead of letrozole with gonadotropins. Reduction of the indicated semaphorins by implantation supports GH's implantation. On the other hand, if this result is supported by new studies, women with implantation-based infertility can provide preference in the selection of drug combinations used for ovulation induction, and can shed light on research for drug development and drug discovery. KEYWORDS: letrozole, growth hormone, gonadotropin, endometrium, semaphorin.