N-Acetylcysteine improves oocyte quality through modulating the Nrf2 signaling pathway to ameliorate oxidative stress caused by repeated controlled ovarian hyperstimulation
Lijie Fan
A B # , Fengli Guan B # , Yucong Ma B , Yu Zhang B , Li Li C , Ying Sun B , Can Cao B , Huilan Du A B * and Ming He B *
+ Author Affiliations
- Author Affiliations
A College of Integrative Medicine, Hebei Medical University, Shijiazhuang, Hebei 050017, China.
B Hebei Key Laboratory of Integrative Medicine on Liver-Kidney Patterns, Institute of Integrative Medicine, College of Integrative Medicine, Hebei University of Chinese Medicine, Shijiazhuang, Hebei 050091, China.
C Reproductive Medicine Center, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
* Correspondence to: duhuilan@hebcm.edu.cn, heming@hebcm.edu.cn
# These authors contributed equally to this paper
Handling Editor: Ye Yuan
Reproduction, Fertility and Development 34(10) 736-750 https://doi.org/10.1071/RD22020
Published online: 6 May 2022
© 2022 The Author(s) (or their employer(s)). Published by CSIRO Publishing. This is an open access article distributed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (CC BY-NC-ND)
Abstract
Context: N-acetyl-cysteine (NAC) is a potent antioxidant that can be used for many gynecological diseases such as polycystic ovary syndrome and endometriosis. Controlled ovarian hyperstimulation (COH) is a critical step in infertility treatment. Our previous clinical studies have shown that repeated COH led to oxidative stress in follicle fluid and ovarian granulosa cells.
Aims: In this study, we investigated whether NAC could inhibit oxidative stress in mice caused by repeated COH and improve the mitochondrial function of oocytes.
Methods: Female Institute of Cancer Research (ICR) mice were randomly assigned into three groups: normal group, model (repeated COH) group, NAC group. We examined the morphology, number and quality of mitochondria. The mechanism of regulation of nuclear factor erythroid 2-related factor 2 (Nrf2) by NAC to ameliorate oxidative stress was also investigated.
Key results: Repeated COH caused oxidative damage in ovaries and oocytes and decreased oocyte quality, while NAC prevented oxidative damage and increased oocyte mitochondrial function. In in vitro experiments, it was verified that NAC can promote the nuclear translocation of Nrf2, which transcriptionally activates the expression of superoxide dismutase and glutathione peroxidase, which removed excessive reactive oxygen species that causes mitochondria damage.
Conclusions: The results suggest that NAC raises mitochondrial function in oocytes and improves oocyte quality through decreasing oxidative stress in mice with repeated COH. The underlying mechanism is related to the regulation of the Nrf2 signaling pathway.
Implication: This study provides a meaningful foundation for the future clinical application of NAC during repeated COH.
Keywords: assisted reproductive technology, controlled ovarian hyperstimulation (COH), infertility, mitochondria, N-acetylcysteine (NAC), Nrf2, oocyte quality, oxidative stress.
References
Adelusi, TI, Du, L, Hao, M, Zhou, X, Xuan, Q, Apu, C, Sun, Y, Lu, Q, and Yin, X (2020). Keap1/Nrf2/are signaling unfolds therapeutic targets for redox imbalanced-mediated diseases and diabetic nephropathy. Biomedicine & Pharmacotherapy 123, 109732.| Keap1/Nrf2/are signaling unfolds therapeutic targets for redox imbalanced-mediated diseases and diabetic nephropathy.Crossref | GoogleScholarGoogle Scholar |
Adeoye, O, Olawumi, J, Opeyemi, A, and Christiania, O (2018). Review on the role of glutathione on oxidative stress and infertility. JBRA Assisted Reproduction 22, 61–66.
| Review on the role of glutathione on oxidative stress and infertility.Crossref | GoogleScholarGoogle Scholar | 29266896PubMed |
Agarwal, A, Aponte-Mellado, A, Premkumar, BJ, Shaman, A, and Gupta, S (2012). The effects of oxidative stress on female reproduction: a review. Reproductive Biology and Endocrinology 10, 49.
| The effects of oxidative stress on female reproduction: a review.Crossref | GoogleScholarGoogle Scholar | 22748101PubMed |
Ambruosi, B, Uranio, MF, Sardanelli, AM, Pocar, P, Martino, NA, Paternoster, MS, Amati, F, and Dell’Aquila, ME (2011). In vitro acute exposure to DEHP affects oocyte meiotic maturation, energy and oxidative stress parameters in a large animal model. PLoS ONE 6, e27452.
| In vitro acute exposure to DEHP affects oocyte meiotic maturation, energy and oxidative stress parameters in a large animal model.Crossref | GoogleScholarGoogle Scholar | 22076161PubMed |
Augoulea, A, Mastorakos, G, Lambrinoudaki, I, Christodoulakos, G, and Creatsas, G (2009). The role of the oxidative-stress in the endometriosis-related infertility. Gynecological Endocrinology 25, 75–81.
| The role of the oxidative-stress in the endometriosis-related infertility.Crossref | GoogleScholarGoogle Scholar | 19253102PubMed |
Babayev, E, and Seli, E (2015). Oocyte mitochondrial function and reproduction. Current Opinion in Obstetrics & Gynecology 27, 175–181.
| Oocyte mitochondrial function and reproduction.Crossref | GoogleScholarGoogle Scholar |
Barrozo, LG, Paulino, LRFM, Silva, BR, Barbalho, EC, Nascimento, DR, Neto, MFL, and Silva, JRV (2021). N-acetyl-cysteine and the control of oxidative stress during in vitro ovarian follicle growth, oocyte maturation, embryo development and cryopreservation. Animal Reproduction Science 231, 106801.
| N-acetyl-cysteine and the control of oxidative stress during in vitro ovarian follicle growth, oocyte maturation, embryo development and cryopreservation.Crossref | GoogleScholarGoogle Scholar | 34252825PubMed |
Bentov, Y, and Casper, RF (2013). The aging oocyte—can mitochondrial function be improved? Fertility and Sterility 99, 18–22.
| The aging oocyte—can mitochondrial function be improved?Crossref | GoogleScholarGoogle Scholar | 23273985PubMed |
Cheraghi, E, Mehranjani, MS, Shariatzadeh, MA, Esfahani, MH, and Ebrahimi, Z (2016). N-acetylcysteine improves oocyte and embryo quality in polycystic ovary syndrome patients undergoing intracytoplasmic sperm injection: an alternative to metformin. Reproduction, Fertility and Development 28, 723–731.
| N-acetylcysteine improves oocyte and embryo quality in polycystic ovary syndrome patients undergoing intracytoplasmic sperm injection: an alternative to metformin.Crossref | GoogleScholarGoogle Scholar |
Combelles, CMH, Cekleniak, NA, Racowsky, C, and Albertini, DF (2002). Assessment of nuclear and cytoplasmic maturation in in-vitro matured human oocytes. Human Reproduction 17, 1006–1016.
| Assessment of nuclear and cytoplasmic maturation in in-vitro matured human oocytes.Crossref | GoogleScholarGoogle Scholar |
Costello, MF, Shrestha, B, Eden, J, Johnson, NP, and Sjoblom, P (2007). Metformin versus oral contraceptive pill in polycystic ovary syndrome: a cochrane review. Human Reproduction 22, 1200–1209.
| Metformin versus oral contraceptive pill in polycystic ovary syndrome: a cochrane review.Crossref | GoogleScholarGoogle Scholar | 17261574PubMed |
Dash, M, Dey, A, and Chattopadhyay, S (2020). Mitigation of arsenic driven utero-ovarian malfunction and changes of apoptotic gene expression by dietary NAC. Ecotoxicology and Environment Safety 199, 110675.
| Mitigation of arsenic driven utero-ovarian malfunction and changes of apoptotic gene expression by dietary NAC.Crossref | GoogleScholarGoogle Scholar |
Dröge, W (2002). Free radicals in the physiological control of cell function. Physiological Reviews 82, 47–95.
| Free radicals in the physiological control of cell function.Crossref | GoogleScholarGoogle Scholar | 11773609PubMed |
Fabbri, R, Sapone, A, Paolini, M, Vivarelli, F, Franchi, P, Lucarini, M, Pasquinelli, G, Vicenti, R, Macciocca, M, Venturoli, S, and Canistro, D (2015). Effects of N-acetylcysteine on human ovarian tissue preservation undergoing cryopreservation procedure. Histology and Histopathology 30, 725–735.
| Effects of N-acetylcysteine on human ovarian tissue preservation undergoing cryopreservation procedure.Crossref | GoogleScholarGoogle Scholar | 25541725PubMed |
Firuzi, O, Miri, R, Tavakkoli, M, and Saso, L (2011). Antioxidant therapy: current status and future prospects. Current Medicinal Chemistry 18, 3871–3888.
| Antioxidant therapy: current status and future prospects.Crossref | GoogleScholarGoogle Scholar | 21824100PubMed |
Galluzzi, L, Kepp, O, Trojel-Hansen, C, and Kroemer, G (2012). Mitochondrial control of cellular life, stress, and death. Circulation Research 111, 1198–1207.
| Mitochondrial control of cellular life, stress, and death.Crossref | GoogleScholarGoogle Scholar | 23065343PubMed |
Hu, X, Roberts, JR, Apopa, PL, Kan, YW, and Ma, Q (2006). Accelerated ovarian failure induced by 4-vinyl cyclohexene diepoxide in Nrf2 null mice. Molecular and Cellular Biology 26, 940–954.
| Accelerated ovarian failure induced by 4-vinyl cyclohexene diepoxide in Nrf2 null mice.Crossref | GoogleScholarGoogle Scholar | 16428448PubMed |
Hu, X, Cheng, L, Wang, X, Luo, G, Zhao, T, Tian, J, and An, L (2020). N-acetyl-l-cysteine protects porcine oocytes undergoing meiotic resumption from heat stress. Reproductive Toxicology 91, 27–34.
| N-acetyl-l-cysteine protects porcine oocytes undergoing meiotic resumption from heat stress.Crossref | GoogleScholarGoogle Scholar | 31698002PubMed |
Iwata, H, Goto, H, Tanaka, H, Sakaguchi, Y, Kimura, K, Kuwayama, T, and Monji, Y (2011). Effect of maternal age on mitochondrial DNA copy number, ATP content and IVF outcome of bovine oocytes. Reproduction, Fertility and Development 23, 424–432.
| Effect of maternal age on mitochondrial DNA copy number, ATP content and IVF outcome of bovine oocytes.Crossref | GoogleScholarGoogle Scholar |
Izzo, CR, Monteleone, PAA, and Serafini, PC (2015). Human reproduction: current status. Revista da Associacao Medica Brasileira 61, 557–559.
| Human reproduction: current status.Crossref | GoogleScholarGoogle Scholar | 26841167PubMed |
Kalthur, G, Salian, SR, Nair, R, Mathew, J, Adiga, SK, Kalthur, SG, Zeegers, D, and Hande, MP (2016). Distribution pattern of cytoplasmic organelles, spindle integrity, oxidative stress, octamer-binding transcription factor 4 (Oct4) expression and developmental potential of oocytes following multiple superovulation. Reproduction, Fertility and Development 28, 2027–2038.
| Distribution pattern of cytoplasmic organelles, spindle integrity, oxidative stress, octamer-binding transcription factor 4 (Oct4) expression and developmental potential of oocytes following multiple superovulation.Crossref | GoogleScholarGoogle Scholar |
Kim, EH, Ridlo, MR, Lee, BC, and Kim, GA (2020). Melatonin-Nrf2 signaling activates peroxisomal activities in porcine cumulus cell-oocyte complexes. Antioxidants 9, 1080.
| Melatonin-Nrf2 signaling activates peroxisomal activities in porcine cumulus cell-oocyte complexes.Crossref | GoogleScholarGoogle Scholar |
Lai, FN, Ma, JY, Liu, JC, Wang, JJ, Cheng, SF, Sun, XF, Li, L, Li, B, Nyachoti, CM, and Shen, W (2015). The influence of N-acetyl-l-cysteine on damage of porcine oocyte exposed to zearalenone in vitro. Toxicology and Applied Pharmacology 289, 341–348.
| The influence of N-acetyl-l-cysteine on damage of porcine oocyte exposed to zearalenone in vitro.Crossref | GoogleScholarGoogle Scholar | 26386189PubMed |
Li, Y, Ruan, X, Liebenthron, J, Montag, M, Zhou, Q, Kong, W, Du, J, Jin, F, Li, S, Cheng, J, Wang, H, and Mueck, AO (2019). Ovarian tissue cryopreservation for patients with premature ovary insufficiency caused by cancer treatment: optimal protocol. Climacteric 22, 383–389.
| Ovarian tissue cryopreservation for patients with premature ovary insufficiency caused by cancer treatment: optimal protocol.Crossref | GoogleScholarGoogle Scholar | 30676094PubMed |
Liu, J, Liu, M, Ye, X, Liu, K, Huang, J, Wang, L, Ji, G, Liu, N, Tang, X, Baltz, JM, Keefe, DL, and Liu, L (2012). Delay in oocyte aging in mice by the antioxidant N-acetyl-l-cysteine (NAC). Human Reproduction 27, 1411–1420.
| Delay in oocyte aging in mice by the antioxidant N-acetyl-l-cysteine (NAC).Crossref | GoogleScholarGoogle Scholar | 22357770PubMed |
Lord, T, and Aitken, RJ (2013). Oxidative stress and ageing of the post-ovulatory oocyte. Reproduction 146, R217–R227.
| Oxidative stress and ageing of the post-ovulatory oocyte.Crossref | GoogleScholarGoogle Scholar | 23950493PubMed |
Ma, R, Liang, W, Sun, Q, Qiu, X, Lin, Y, Ge, X, Jueraitetibaike, K, Xie, M, Zhou, J, Huang, X, Wang, Q, Chen, L, and Yao, B (2018). Sirt1/Nrf2 pathway is involved in oocyte aging by regulating cyclin B1. Aging 10, 2991–3004.
| Sirt1/Nrf2 pathway is involved in oocyte aging by regulating cyclin B1.Crossref | GoogleScholarGoogle Scholar | 30368232PubMed |
Ma, Y, Zhao, Z, Hao, G, Cui, N, Fan, Y, Cao, Y, Tan, Z, Geng, J, Fan, L, Du, H, and Gao, B (2021). Effects of multicycle gonadotropin-releasing hormone antagonist protocols on oxidative stress of follicular fluid and ovarian granulosa cells. Human Cell 34, 1324–1334.
| Effects of multicycle gonadotropin-releasing hormone antagonist protocols on oxidative stress of follicular fluid and ovarian granulosa cells.Crossref | GoogleScholarGoogle Scholar | 33963989PubMed |
Mahmoodi, M, Soleimani Mehranjani, M, Shariatzadeh, SM, Eimani, H, and Shahverdi, A (2015). N-acetylcysteine improves function and follicular survival in mice ovarian grafts through inhibition of oxidative stress. Reproductive BioMedicine Online 30, 101–110.
| N-acetylcysteine improves function and follicular survival in mice ovarian grafts through inhibition of oxidative stress.Crossref | GoogleScholarGoogle Scholar | 25458850PubMed |
Martin-Johnston, MK, Uhler, ML, Grotjan, HE, Lifchez, AS, Nani, JM, and Beltsos, AN (2009). Lower chance of pregnancy with repeated cycles with in vitro fertilization. Journal of Reproductive medicine 54, 67–72.
May-Panloup, P, Chrétien, MF, Jacques, C, Vasseur, C, Malthiéry, Y, and Reynier, P (2005). Low oocyte mitochondrial DNA content in ovarian insufficiency. Human Reproduction 20, 593–597.
| Low oocyte mitochondrial DNA content in ovarian insufficiency.Crossref | GoogleScholarGoogle Scholar | 15608038PubMed |
Mikolka, P, Kopincova, J, Mikusiakova, LT, Kosutova, P, Calkovska, A, and Mokra, D (2016). Antiinflammatory effect of N-acetylcysteine combined with exogenous surfactant in meconium-induced lung injury. Advances in Experimental Medicine and Biology 934, 63–75.
| Antiinflammatory effect of N-acetylcysteine combined with exogenous surfactant in meconium-induced lung injury.Crossref | GoogleScholarGoogle Scholar | 27283193PubMed |
Miyamoto, K, Sato, EF, Kasahara, E, Jikumaru, M, Hiramoto, K, Tabata, H, Katsuragi, M, Odo, S, Utsumi, K, and Inoue, M (2010). Effect of oxidative stress during repeated ovulation on the structure and functions of the ovary, oocytes, and their mitochondria. Free Radical Biology and Medicine 49, 674–681.
| Effect of oxidative stress during repeated ovulation on the structure and functions of the ovary, oocytes, and their mitochondria.Crossref | GoogleScholarGoogle Scholar | 20621580PubMed |
Pei, Y, Liu, H, Yang, Y, Yang, Y, Jiao, Y, Tay, FR, and Chen, J (2018). Biological activities and potential oral applications of N-acetylcysteine: progress and prospects. Oxidative Medicine and Cellular Longevity 2018, 2835787.
| Biological activities and potential oral applications of N-acetylcysteine: progress and prospects.Crossref | GoogleScholarGoogle Scholar | 29849877PubMed |
Salehpour, S, Akbari Sene, A, Saharkhiz, N, Sohrabi, MR, and Moghimian, F (2012). N-acetylcysteine as an adjuvant to clomiphene citrate for successful induction of ovulation in infertile patients with polycystic ovary syndrome. Journal of Obstetrics and Gynaecology Research 38, 1182–1186.
| N-acetylcysteine as an adjuvant to clomiphene citrate for successful induction of ovulation in infertile patients with polycystic ovary syndrome.Crossref | GoogleScholarGoogle Scholar |
Sanchez, F, Romero, S, De Vos, M, Verheyen, G, and Smitz, J (2015). Human cumulus-enclosed germinal vesicle oocytes from early antral follicles reveal heterogeneous cellular and molecular features associated with in vitro maturation capacity. Human Reproduction 30, 1396–1409.
| Human cumulus-enclosed germinal vesicle oocytes from early antral follicles reveal heterogeneous cellular and molecular features associated with in vitro maturation capacity.Crossref | GoogleScholarGoogle Scholar | 25904637PubMed |
Schieber, M, and Chandel, NS (2014). ROS function in redox signaling and oxidative stress. Current Biology 24, R453–R462.
| ROS function in redox signaling and oxidative stress.Crossref | GoogleScholarGoogle Scholar | 24845678PubMed |
Shapiro, BS, Richter, KS, Harris, DC, and Daneshmand, ST (2001). Dramatic declines in implantation and pregnancy rates in patients who undergo repeated cycles of in vitro fertilization with blastocyst transfer after one or more failed attempts. Fertility and Sterility 76, 538–542.
| Dramatic declines in implantation and pregnancy rates in patients who undergo repeated cycles of in vitro fertilization with blastocyst transfer after one or more failed attempts.Crossref | GoogleScholarGoogle Scholar | 11532478PubMed |
Sharma, M, Kaur, T, and Singla, SK (2016). Role of mitochondria and NADPH oxidase derived reactive oxygen species in hyperoxaluria induced nephrolithiasis: therapeutic intervention with combinatorial therapy of N-acetyl cysteine and apocynin. Mitochondrion 27, 15–24.
| Role of mitochondria and NADPH oxidase derived reactive oxygen species in hyperoxaluria induced nephrolithiasis: therapeutic intervention with combinatorial therapy of N-acetyl cysteine and apocynin.Crossref | GoogleScholarGoogle Scholar | 26779823PubMed |
Shaw, P, and Chattopadhyay, A (2020). Nrf2-are signaling in cellular protection: mechanism of action and the regulatory mechanisms. Journal of Cellular Physiology 235, 3119–3130.
| Nrf2-are signaling in cellular protection: mechanism of action and the regulatory mechanisms.Crossref | GoogleScholarGoogle Scholar | 31549397PubMed |
Shigenaga MK, Hagen TM, Ames BN (1994) Oxidative damage and mitochondrial decay in aging. Proceedings of the National Academy of Sciences of the United States of America 91, 10771–10778.
| Crossref |
Smith, ADAC, Tilling, K, Nelson, SM, and Lawlor, DA (2015). Live-birth rate associated with repeat in vitro fertilization treatment cycles. JAMA 314, 2654–2662.
| Live-birth rate associated with repeat in vitro fertilization treatment cycles.Crossref | GoogleScholarGoogle Scholar | 26717030PubMed |
Sohel, MMH, Akyuz, B, Konca, Y, Arslan, K, Sariozkan, S, and Cinar, MU (2019). Oxidative stress modulates the expression of apoptosis-associated microRNAs in bovine granulosa cells in vitro. Cell and Tissue Research 376, 295–308.
| Oxidative stress modulates the expression of apoptosis-associated microRNAs in bovine granulosa cells in vitro.Crossref | GoogleScholarGoogle Scholar | 30666538PubMed |
Subramanian, R, Tam, J, Aidasani, D, Reid, DL, and Skiles, GL (2011). Novel cytochrome p450 bioactivation of a terminal phenyl acetylene group: formation of a one-carbon loss benzaldehyde and other oxidative products in the presence of N-acetyl cysteine or glutathione. Chemical Research in Toxicology 24, 677–686.
| Novel cytochrome p450 bioactivation of a terminal phenyl acetylene group: formation of a one-carbon loss benzaldehyde and other oxidative products in the presence of N-acetyl cysteine or glutathione.Crossref | GoogleScholarGoogle Scholar | 21395287PubMed |
Torner, H, Brüssow, K-P, Alm, H, Ratky, J, Pöhland, R, Tuchscherer, A, and Kanitz, W (2004). Mitochondrial aggregation patterns and activity in porcine oocytes and apoptosis in surrounding cumulus cells depends on the stage of pre-ovulatory maturation. Theriogenology 61, 1675–1689.
| Mitochondrial aggregation patterns and activity in porcine oocytes and apoptosis in surrounding cumulus cells depends on the stage of pre-ovulatory maturation.Crossref | GoogleScholarGoogle Scholar | 15019463PubMed |
Van Blerkom, J, and Davis, P (2007). Mitochondrial signaling and fertilization. Molecular Human Reproduction 13, 759–770.
| Mitochondrial signaling and fertilization.Crossref | GoogleScholarGoogle Scholar | 17893093PubMed |
Van Blerkom, J, Davis, PW, and Lee, J (1995). ATP content of human oocytes and developmental potential and outcome after in-vitro fertilization and embryo transfer. Human Reproduction 10, 415–424.
| ATP content of human oocytes and developmental potential and outcome after in-vitro fertilization and embryo transfer.Crossref | GoogleScholarGoogle Scholar | 7769073PubMed |
Wakabayashi, N, Slocum, SL, Skoko, JJ, Shin, S, and Kensler, TW (2010). When NRF2 talks, who’s listening? Antioxidants & Redox Signaling 13, 1649–1663.
| When NRF2 talks, who’s listening?Crossref | GoogleScholarGoogle Scholar |
Wang, L, Zhang, J, Zhao, C, Jia, Z, and Feng, X (2021). Melatonin reverses 10-hydroxycamptothecin-induced apoptosis and autophagy in mouse oocyte. Reproductive Sciences 28, 1839–1849.
| Melatonin reverses 10-hydroxycamptothecin-induced apoptosis and autophagy in mouse oocyte.Crossref | GoogleScholarGoogle Scholar | 33104985PubMed |
Yamada, M, Ueno, T, Minamikawa, H, Sato, N, Iwasa, F, Hori, N, and Ogawa, T (2010). N-acetyl cysteine alleviates cytotoxicity of bone substitute. Journal of Dental Research 89, 411–416.
| N-acetyl cysteine alleviates cytotoxicity of bone substitute.Crossref | GoogleScholarGoogle Scholar | 20200411PubMed |
Zhao, F, Li, K, Zhao, L, Liu, J, Suo, Q, Zhao, J, Wang, H, and Zhao, S (2014). Effect of Nrf2 on rat ovarian tissues against atrazine-induced anti-oxidative response. International Journal of Clinical & Experimental Pathology 7, 2780–2789.
