Document Type : Original Article
Authors
1
.. Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
2
.Department of Infertility, Shariati Hospital, Tehran University of Medical Sciences, Tehran, Iran
3
.Department of Molecular Medicine, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran
4
4.Islamic Azad University, Science and Research Branch, Tehran, Iran
5
.. Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran;5.Department of ART, Embryology Laboratory, Arash Women’s Hospital, Tehran University of Medical Sciences, Tehran, Iran
6
6.Fertility and Infertility Research Center, Health Technology Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran
7
.. Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran;.Department of Infertility, Shariati Hospital, Tehran University of Medical Sciences, Tehran, Iran
Abstract
Objective
Sulforaphane (SFN) is a natural free radical scavenger that can reduce oxidative stress (OS) through mediating nuclear factor (erythroid-derived 2)-like 2 (NF-E2-related factor 2 or NRF2)/antioxidant response element (ARE) signaling pathway and the downstream antioxidant enzymes. Here, we intended to study the role of SFN in OS- induced human granulosa cells (GCs) by investigating the intracellular levels of reactive oxygen species (ROS), cell death, and NRF2-ARE pathway.
Materials and Methods
This experimental study was conducted on GCs of 12 healthy women who had normal menstrual cycles with no history of polycystic ovary syndrome (PCOS), endometriosis, menstrual disorders, hyperprolactinemia, or hormonal therapy. After isolation of GCs, the MTT assay was performed to explore GCs viability after treatment with SFN in the presence or absence of H2O2. Flow cytometry was utilized to determine the intracellular ROS production and the apoptosis rate. Evaluation of the mRNA and protein expression levels of NRF2 and phase II enzymes including superoxide dismutase (SOD) and catalase (CAT) was performed by quantitative real-time polymerase chain reaction (PCR) and western blotting. Finally, the data were analyzed by SPSS software using One-way ANOVA and the suitable post-hoc test. Significance level was considered as P < 0.05.
Results
Pretreatment of GCs with SFN attenuated intracellular ROS production and apoptosis rate in the H2O2-exposed cells. Moreover, SFN treatment increased the mRNA expression level of NRF2, SOD, and CAT. Higher expression of NRF2 and SOD was also observed at the protein level.
Conclusion
Our study demonstrated that SFN protects human GCs against H2O2induced-OS by reducing the intracellular ROS production and the following apoptosis through a mechanism by which NRF2 increases the antioxidant enzymes such as SOD and CAT. This result may have a potential application in assisted reproduction cycles by improving the quality of GCs and the embedded oocyte, especially in PCOS patients.
Keywords
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