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

Overview

Journal Cell J

Specialty Cell Biology

Date 2021 Jul 26

PMID 34308575

Citations 12

Authors

Mojtaba Eslami

Sahar Esfandyari

Marzieh Aghahosseini

Zahra Rashidi

S Hirzad Hosseinishental

Samane Brenjian

Aligholi Sobhani

Fardin Amidi

Affiliations

Soon will be listed here.

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 (HO) 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 fertilization (IVF) outcomes and treatment of infertility.

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References

Yoshihisa Y, Rehman M, Shimizu T . Astaxanthin, a xanthophyll carotenoid, inhibits ultraviolet-induced apoptosis in keratinocytes. Exp Dermatol. 2014; 23(3):178-83. DOI: 10.1111/exd.12347. View

Zhu X, Chen Y, Chen Q, Yang H, Xie X . Astaxanthin Promotes Nrf2/ARE Signaling to Alleviate Renal Fibronectin and Collagen IV Accumulation in Diabetic Rats. J Diabetes Res. 2018; 2018:6730315. PMC: 5884145. DOI: 10.1155/2018/6730315. View

Li Z, Dong X, Liu H, Chen X, Shi H, Fan Y . Astaxanthin protects ARPE-19 cells from oxidative stress via upregulation of Nrf2-regulated phase II enzymes through activation of PI3K/Akt. Mol Vis. 2013; 19:1656-66. PMC: 3725964. View

Davinelli S, Nielsen M, Scapagnini G . Astaxanthin in Skin Health, Repair, and Disease: A Comprehensive Review. Nutrients. 2018; 10(4). PMC: 5946307. DOI: 10.3390/nu10040522. View

Mahmood T, Yang P . Western blot: technique, theory, and trouble shooting. N Am J Med Sci. 2012; 4(9):429-34. PMC: 3456489. DOI: 10.4103/1947-2714.100998. View

Saw C, Yang A, Guo Y, Kong A . Astaxanthin and omega-3 fatty acids individually and in combination protect against oxidative stress via the Nrf2-ARE pathway. Food Chem Toxicol. 2013; 62:869-75. DOI: 10.1016/j.fct.2013.10.023. View

Karuputhula N, Chattopadhyay R, Chakravarty B, Chaudhury K . Oxidative status in granulosa cells of infertile women undergoing IVF. Syst Biol Reprod Med. 2013; 59(2):91-8. DOI: 10.3109/19396368.2012.743197. View

Jia Y, Lin J, Mi Y, Zhang C . Quercetin attenuates cadmium-induced oxidative damage and apoptosis in granulosa cells from chicken ovarian follicles. Reprod Toxicol. 2011; 31(4):477-85. DOI: 10.1016/j.reprotox.2010.12.057. View

Devine P, Perreault S, Luderer U . Roles of reactive oxygen species and antioxidants in ovarian toxicity. Biol Reprod. 2011; 86(2):27. PMC: 3290661. DOI: 10.1095/biolreprod.111.095224. View

10.

Van Meerloo J, Kaspers G, Cloos J . Cell sensitivity assays: the MTT assay. Methods Mol Biol. 2011; 731:237-45. DOI: 10.1007/978-1-61779-080-5_20. View

11.

Akino N, Wada-Hiraike O, Terao H, Honjoh H, Isono W, Fu H . Activation of Nrf2 might reduce oxidative stress in human granulosa cells. Mol Cell Endocrinol. 2017; 470:96-104. DOI: 10.1016/j.mce.2017.10.002. View

12.

Eggler A, Gay K, Mesecar A . Molecular mechanisms of natural products in chemoprevention: induction of cytoprotective enzymes by Nrf2. Mol Nutr Food Res. 2008; 52 Suppl 1:S84-94. DOI: 10.1002/mnfr.200700249. View

13.

Sawai H, Domae N . Discrimination between primary necrosis and apoptosis by necrostatin-1 in Annexin V-positive/propidium iodide-negative cells. Biochem Biophys Res Commun. 2011; 411(3):569-73. DOI: 10.1016/j.bbrc.2011.06.186. View

14.

Kubista M, Andrade J, Bengtsson M, Forootan A, Jonak J, Lind K . The real-time polymerase chain reaction. Mol Aspects Med. 2006; 27(2-3):95-125. DOI: 10.1016/j.mam.2005.12.007. View

15.

Rashidi Z, Aleyasin A, Eslami M, Nekoonam S, Zendedel A, Bahramrezaie M . Quercetin protects human granulosa cells against oxidative stress via thioredoxin system. Reprod Biol. 2019; 19(3):245-254. DOI: 10.1016/j.repbio.2019.07.002. View

16.

Niu T, Xuan R, Jiang L, Wu W, Zhen Z, Song Y . Astaxanthin Induces the Nrf2/HO-1 Antioxidant Pathway in Human Umbilical Vein Endothelial Cells by Generating Trace Amounts of ROS. J Agric Food Chem. 2018; 66(6):1551-1559. DOI: 10.1021/acs.jafc.7b05493. View

17.

Furfaro A, Traverso N, Domenicotti C, Piras S, Moretta L, Marinari U . The Nrf2/HO-1 Axis in Cancer Cell Growth and Chemoresistance. Oxid Med Cell Longev. 2015; 2016:1958174. PMC: 4677237. DOI: 10.1155/2016/1958174. View

18.

Lee D, Kim C, Lee Y . Astaxanthin protects against MPTP/MPP+-induced mitochondrial dysfunction and ROS production in vivo and in vitro. Food Chem Toxicol. 2010; 49(1):271-80. PMC: 3010303. DOI: 10.1016/j.fct.2010.10.029. View

19.

Wu Q, Zhang X, Wang H, Zhang X, Yu Q, Li W . Astaxanthin activates nuclear factor erythroid-related factor 2 and the antioxidant responsive element (Nrf2-ARE) pathway in the brain after subarachnoid hemorrhage in rats and attenuates early brain injury. Mar Drugs. 2014; 12(12):6125-41. PMC: 4278222. DOI: 10.3390/md12126125. View

20.

Upadhyay S, Dixit M . Role of Polyphenols and Other Phytochemicals on Molecular Signaling. Oxid Med Cell Longev. 2015; 2015:504253. PMC: 4477245. DOI: 10.1155/2015/504253. View