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Quercetin Protects Human Granulosa Cells Against Oxidative Stress Via Thioredoxin System Publisher Pubmed



Rashidi Z1 ; Aleyasin A2 ; Eslami M1 ; Nekoonam S1 ; Zendedel A3 ; Bahramrezaie M4 ; Amidi F1, 2
Authors
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Authors Affiliations
  1. 1. Department of Anatomy, School of Medicine, Tehran Universityof Medical Sciences, Pour Sina St, Tehran, 1417613151, Iran
  2. 2. Department of Infertility, Shariati Hospital, Tehran University of Medical Sciences, 1411713135, Tehran, Iran
  3. 3. Institute of Neuroanatomy, Faculty of Medicine, RWTH Aachen University, 52062, Germany
  4. 4. Cellular and Molecular Research Center, Research Institute for Health Development, Kurdistan University of Medical Sciences, Sanandaj, Iran

Source: Reproductive Biology Published:2019


Abstract

Granulosa Cells (GCs) are sensitive to excessive production of reactive oxygen species (ROS). Quercetin (QUR) is a free radical scavenger which can alleviate oxidative stress through nuclear factor (erythroid-derived 2)-like 2 (Nrf2)/antioxidant response element (ARE) pathway and thioredoxin (Trx) system. We aimed to explore the probable protective role of QUR on cultured human GCs treated with hydrogen peroxide (H2O2) as an inducer of oxidative stress. MTT assay was applied for evaluating the cell cytotoxicity of QUR and H2O2. The rate of apoptotic cells and intracellular ROS generation were determined by Annexin V-FITC/PI staining and 2′-7′-dichlorodihydrofluorescein diacetate fluorescent probes (DCFH-DA), respectively. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis and western blot analysis were used to evaluate the gene and protein expression of Nrf2 and kelch-like ech-associated protein 1 (Keap1)1. The Nrf2 and Trx activities were measured by Enzyme-linked Immunosorbent Assay (ELISA). The results indicated that QUR pretreatment can decrease ROS production and apoptosis induced by H2O2. In addition, QUR increased Nrf2 gene and protein expression, as well as its nuclear translocation. Moreover, in QUR-treated group, a lower level of Keap1 protein was observed, which was not reported as significant. The results also indicated a significant correlation between the expression of Nrf2 and Keap1 in QUR-treated group. Further, QUR protected GCs from oxidative stress by increasing Trx gene expression and activity. This study suggests that QUR as a supplementary factor may protect GCs from oxidative stress in diseases related to this condition. © 2019 Society for Biology of Reproduction & the Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn
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