Astaxanthin Protects Human Granulosa Cells against Oxidative Stress through Activation of NRF2/ARE Pathway and Its Downstream Phase II Enzymes
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 exami...
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| Format: | Article |
| Language: | English |
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Royan Institute (ACECR), Tehran
2021-08-01
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| Series: | Cell Journal |
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| Online Access: | https://celljournal.org/journal/article/fulltext/astaxanthin-protects-human-granulosa-cells-against-oxidative-stress-through-activation-of-nrf2are-pathway-and-its-downstream-phase-ii-enzymes.pdf |
| _version_ | 1819141961563504640 |
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| author | Mojtaba Eslami Sahar Esfandyari Marzieh Aghahosseini Zahra Rashidi Shirzad Hosseinishental Samane Brenjian Aligholi Sobhani Fardin Amidi |
| author_facet | Mojtaba Eslami Sahar Esfandyari Marzieh Aghahosseini Zahra Rashidi Shirzad Hosseinishental Samane Brenjian Aligholi Sobhani Fardin Amidi |
| author_sort | Mojtaba Eslami |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-22T12:02:46Z |
| format | Article |
| id | doaj.art-fcb8fec66b8e4a3398e6f4966df7d49e |
| institution | Directory Open Access Journal |
| issn | 2228-5806 2228-5814 |
| language | English |
| last_indexed | 2024-12-22T12:02:46Z |
| publishDate | 2021-08-01 |
| publisher | Royan Institute (ACECR), Tehran |
| record_format | Article |
| series | Cell Journal |
| spelling | doaj.art-fcb8fec66b8e4a3398e6f4966df7d49e2022-12-21T18:26:33ZengRoyan Institute (ACECR), TehranCell Journal2228-58062228-58142021-08-0123331932810.22074/cellj.2021.7222Astaxanthin Protects Human Granulosa Cells against Oxidative Stress through Activation of NRF2/ARE Pathway and Its Downstream Phase II EnzymesMojtaba Eslami0Sahar Esfandyari1Marzieh Aghahosseini2Zahra Rashidi3Shirzad Hosseinishental4Samane Brenjian5Aligholi Sobhani6Fardin Amidi7Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, IranDepartment of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, IranDepartment of Infertility, Shariati Hospital, Tehran University of Medical Sciences, Tehran, IranFertility and Infertility Research Center, Health Technology Institute, Kermanshah University of Medical Sciences, Kermanshah, IranDepartment of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, IranDepartment of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, IranDepartment of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, IranDepartment of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, IranObjective: 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.https://celljournal.org/journal/article/fulltext/astaxanthin-protects-human-granulosa-cells-against-oxidative-stress-through-activation-of-nrf2are-pathway-and-its-downstream-phase-ii-enzymes.pdfastaxanthingranulosa cellsnuclear factor-e2-related factor 2oxidative stress |
| spellingShingle | Mojtaba Eslami Sahar Esfandyari Marzieh Aghahosseini Zahra Rashidi Shirzad Hosseinishental Samane Brenjian Aligholi Sobhani Fardin Amidi Astaxanthin Protects Human Granulosa Cells against Oxidative Stress through Activation of NRF2/ARE Pathway and Its Downstream Phase II Enzymes Cell Journal astaxanthin granulosa cells nuclear factor-e2-related factor 2 oxidative stress |
| title | Astaxanthin Protects Human Granulosa Cells against Oxidative Stress through Activation of NRF2/ARE Pathway and Its Downstream Phase II Enzymes |
| title_full | Astaxanthin Protects Human Granulosa Cells against Oxidative Stress through Activation of NRF2/ARE Pathway and Its Downstream Phase II Enzymes |
| title_fullStr | Astaxanthin Protects Human Granulosa Cells against Oxidative Stress through Activation of NRF2/ARE Pathway and Its Downstream Phase II Enzymes |
| title_full_unstemmed | Astaxanthin Protects Human Granulosa Cells against Oxidative Stress through Activation of NRF2/ARE Pathway and Its Downstream Phase II Enzymes |
| title_short | Astaxanthin Protects Human Granulosa Cells against Oxidative Stress through Activation of NRF2/ARE Pathway and Its Downstream Phase II Enzymes |
| title_sort | astaxanthin protects human granulosa cells against oxidative stress through activation of nrf2 are pathway and its downstream phase ii enzymes |
| topic | astaxanthin granulosa cells nuclear factor-e2-related factor 2 oxidative stress |
| url | https://celljournal.org/journal/article/fulltext/astaxanthin-protects-human-granulosa-cells-against-oxidative-stress-through-activation-of-nrf2are-pathway-and-its-downstream-phase-ii-enzymes.pdf |
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