The Mechanism Exploration of Follicular Fluids on Granulose Cell Apoptosis in Endometriosis-Associated Infertility.

Lu Chen,Zailong Cai,Zhexin Ni,Chaoqin Yu,Jinyu Jinyu,Wen Cheng,Shuai Sun,Hai-Feng Pan

doi:10.1155/2021/6464686

Abstract

Objective To explore the mechanisms of follicular fluids (FFs) on granulose cell (GC) apoptosis in endometriosis-associated infertility. Materials and Methods 60 infertile women were enrolled. The FFs from 30 endometriosis-associated infertility (EI) patients were collected and processed by ELISA hormone assay and proteomic profiling. The ovary GCs collected from 30 tubal-associated infertility (TI) patients were cultured in follicular fluids of endometriosis-associated infertility patients (EI-FFs), and the apoptosis mechanisms were explored by flow cytometry assay, real-time PCR, Western blotting, and protein–protein interaction (PPI) network analysis. Results Our results showed that the expression of 22 specific proteins was significantly different in the FFs from EI and TI patients, and the level of testosterone and anti-Müllerian hormone was not obviously different between the two groups. EI-FFs could accelerate the apoptosis process of granulose cells of tubal-associated infertility patients (TI-GCs) by regulating the expression of 5 apoptosis-related proteins including BCL2, BAX, CASP3, CASP9, and TP53. The correlation of these 22 specific proteins and 5 apoptosis-related proteins was analyzed by PPI, and 5 protein biomarkers (INS, CXCL10, ICAM1, WIF1, and TNFRSF13C) and 5 signaling pathways (cytokine-cytokine receptor interaction, apoptosis, regulation of actin cytoskeleton, MAPK, and p53 signaling pathway) were predicted. Conclusion This research clarified the effect and explored the mechanisms of EI-FFs on the apoptosis of TI-GCs and indicated the protein biomarkers and signaling pathways for further study.

Highlights

Our study indicated that 9% endometriosis-associated infertility (EI)-Follicular fluids (FFs) intervention in tubal-associated infertility (TI)-granulose cell (GC) for 48 h had an obvious effect on apoptosis (Figure 4), and the apoptosis rate was significantly increased compared with 9% TI-FFs on TI-GCs (Figure 5)
The results indicated that EI-FFs induced the apoptosis of TI-GCs by regulating multiple biological processes (p < 0:01), and the top five of them were positive regulation of protein metabolic process (GO:0051247), cell surface receptor signaling pathway (GO:0007166), regulation of response to stimulus (GO:0048583), positive regulation of signaling (GO:0023056), and positive regulation of cell communication (GO:0010647) (Figure 8(a))
10%–15% of couples at reproductive age suffer from infertility, and a better understanding of the regulatory processes of reproduction could allow for the continuous improvement in the success of infertility treatment [15]

Summary

Introduction

Endometriosis is a common dynamic complex estrogendependent gynecological disorder characterized by the presence of functional endometrial-type mucosa outside the uterine cavity that affects up to 50% of infertile women of reproductive age [1]. Assisted Reproductive Technology (ART) is the main treatment for endometriosis-associated infertility, the number of eggs obtained and the rate of fertilization and pregnancy are lower compared with the nonendometriosis patients. In addition to pelvic structure changes and immune deficiency, the altered follicular microenvironment in endometriosis patients is one of the most important contributing factors to infertility [2]. Granulosa cells (GCs), which surround the oocytes to form the follicular structure, regulate the occurrence, growth, maturation, and atresia of the follicle. The morphological and functional changes of GCs are important markers of follicular development.

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Results

Conclusion