Astaxanthin Protects Human Granulosa Cells against Oxidative Stress through Activation of NRF2/ARE Pathway and Its Downstream Phase II Enzymes

(Pages: 319-328)
Mojtaba Eslami, Ph.D., 1Sahar Esfandyari, D.V.M, 1Marzieh Aghahosseini, M.D., 2Zahra Rashidi, Ph.D., 3Shirzad Hosseinishental, Ph.D., 1Samane Brenjian, Ph.D., 1Aligholi Sobhani, Ph.D., 1Fardin Amidi, Ph.D., 1,*
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
Fertility and Infertility Research Center, Health Technology institute, Kermanshah University of Medical Sciences, Kermanshah, Iran
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
Fertility and Infertility Research Center, Health Technology institute, Kermanshah University of Medical Sciences, Kermanshah, Iran
*Corresponding Address: Department of Anatomy School of Medicine Tehran University of Medical Sciences Tehran Iran Email:famidi@sina.tums.ac.ir
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Eslami Mojtaba, Esfandyari Sahar, Aghahosseini Marzieh, Rashidi Zahra, Hosseinishental Shirzad, Brenjian Samane, Sobhani Aligholi, Amidi Fardin. Astaxanthin Protects Human Granulosa Cells against Oxidative Stress through Activation of NRF2/ARE Pathway and Its Downstream Phase II Enzymes . Cell J. 2021; 23(3): 319-328.

Abstract

Objective

Astaxanthin (AST) has been introduced as a radical scavenger and an anti-apoptotic factor that acts via regulating the nuclear factor-E2-related factor 2 (NRF2) and related factors. Here, we intended to examine the effect of AST on granulosa cells (GCs) against oxidative stress by examining NRF2 and downstream phase II antioxidant enzymes.

Materials and Methods

In this experimental study, we used cultured human primary GCs for the study. First, we performed the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test to evaluate cells viability after treatment with hydrogen peroxide (H2O2) and AST. The apoptosis rate and ROS levels were measured by flow cytometry. To determine NRF2 and phase II enzymes expression, we performed real-time polymerase chain reaction (PCR). Finally, we used western blot to measure the protein levels of NRF2 and Kelch-like ECsH-associated protein 1 (KEAP1). Enzyme activity analysis was also performed to detect NRF2 activity.

Results

This study showed that AST suppressed ROS generation (P<0.01) and cell death (P<0.05) in GCs induced by oxidative stress. AST also elevated gene and protein expression and nuclear localization of NRF2 and had an inhibitory effect on the protein levels of KEAP1 (P<0.05). Furthermore, when we used trigonelline (Trig) as a known inhibitor of NRF2, it attenuated the protective effects of AST by decreasing NRF2 activity and gene expression of phase II enzymes (P<0.05).

Conclusion

Our results presented the protective role of AST against oxidative stress in GCs which was mediated through up-regulating the phase II enzymes as a result of NRF2 activation. Our study may help in improving in vitro fertilization (IVF) outcomes and treatment of infertility.