Nrf2-mediated ferroptosis of spermatogenic cells involved in male reproductive toxicity induced by polystyrene nanoplastics in mice

Xufeng FU; Hang HAN; Hong YANG; Bo XU; Wenjie DAI; Ling LIU; Tiantian HE; Xing DU; Xiuying PEI

doi:10.1631/jzus.B2300138

Your Location：

Home >

Browse articles >

Nrf2-mediated ferroptosis of spermatogenic cells involved in male reproductive toxicity induced by polystyrene nanoplastics in mice

Scan for full text

Alert me when the article has been cited

Submit

Research Article | Updated：2024-04-07

Nrf2-mediated ferroptosis of spermatogenic cells involved in male reproductive toxicity induced by polystyrene nanoplastics in mice
Nrf2-mediated ferroptosis of spermatogenic cells involved in male reproductive toxicity induced by polystyrene nanoplastics in mice
Nrf2介导的生精细胞铁死亡参与了聚苯乙烯纳米塑料导致的小鼠雄性生殖毒性
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology) Vol. 25, Issue 4, Pages: 307-323(2024)
Affiliations：

1.Key Laboratory of Fertility Preservation and Maintenance of Ministry of Education, School of Basic Medical Sciences, Ningxia Medical University, Yinchuan 750004, China
2.Key Laboratory of Reproduction and Genetics in Ningxia, Ningxia Medical University, Yinchuan 750004, China
Funds：
DOI：10.1631/jzus.B2300138
CLC：
CSTR：
Published： 15 April 2024 ，

Received： 28 February 2023 ，

Revised： 29 July 2023 ，

付旭锋,韩杭,杨宏等.Nrf2介导的生精细胞铁死亡参与了聚苯乙烯纳米塑料导致的小鼠雄性生殖毒性[J].浙江大学学报（英文版）（B辑：生物医学和生物技术）,2024,25(04):307-323.

Xufeng FU, Hang HAN, Hong YANG, et al. Nrf2-mediated ferroptosis of spermatogenic cells involved in male reproductive toxicity induced by polystyrene nanoplastics in mice. [J]. Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology) 25(4):307-323(2024)
付旭锋,韩杭,杨宏等.Nrf2介导的生精细胞铁死亡参与了聚苯乙烯纳米塑料导致的小鼠雄性生殖毒性[J].浙江大学学报（英文版）（B辑：生物医学和生物技术）,2024,25(04):307-323. DOI： 10.1631/jzus.B2300138.

Xufeng FU, Hang HAN, Hong YANG, et al. Nrf2-mediated ferroptosis of spermatogenic cells involved in male reproductive toxicity induced by polystyrene nanoplastics in mice. [J]. Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology) 25(4):307-323(2024) DOI： 10.1631/jzus.B2300138.

摘要

由于大量塑料废弃物的排放和一次性口罩的广泛使用，所产生的微塑料（MPs）和纳米塑料（NPs）已被认为是有害物质，但它们对健康的具体影响仍不确定。本研究将荧光标记的聚苯乙烯纳米塑料（PS-NPs）注射到小鼠体内以确定NPs在体内的分布和潜在的毒性作用，通过动物活体成像发现PS-NPs在小鼠睾丸中有明显积累。因此，本文研究了PS-NPs对雄性小鼠生殖系统和对生精细胞的毒性作用及机制。通过雄性小鼠灌胃暴露50 nm和90 nm的PS-NPs后，其生精能力受到影响且生精细胞受损；在体外暴露发现，PS-NPs会影响精母细胞系GC-2的存活；利用RNA-seq进一步分析其毒理机制，发现PS-NPs通过铁死亡途径影响GC-2细胞；通过线粒体形态、Fe

水平、脂质过氧化、线粒体膜电位和不稳定铁等方面评价了PS-NPs引起GC-2细胞铁死亡的表型，进一步明确铁死亡抑制剂Fer-1可以逆转铁死亡表型。随后，发现Nrf2在PS-NPs诱导GC-2细胞铁死亡中起重要作用，并且抑制Nrf2后可加剧PS-NPs诱导的GC-2细胞铁死亡。最后，通过体内实验进一步证实了Nrf2在PS-NPs诱导的雄性生殖毒性中发挥保护作用。因此，本研究表明，PS-NPs通过引起Nrf2介导的生精细胞铁死亡进而导致小鼠雄性生殖功能障碍。

Abstract

Microplastics (MPs) and nanoplastics (NPs) have become hazardous materials due to the massive amount of plastic waste and disposable masks

but their specific health effects remain uncertain. In this study

fluorescence-labeled polystyrene NPs (PS-NPs) were injected into the circulatory systems of mice to determine the distribution and potential toxic effects of NPs in vivo. Interestingly

whole-body imaging found that PS-NPs accumulated in the testes of mice. Therefore

the toxic effects of PS-NPs on the reproduction systems and the spermatocytes cell line of male mice

and their mechanisms

were investigated. After oral exposure to PS-NPs

their spermatogenesis was affected and the spermatogenic cells were damaged. The spermatocyte cell line GC-2 was exposed to PS-NPs and analyzed using RNA sequencing (RNA-seq) to determine the toxic mechanisms; a ferroptosis pathway was found after PS-NP exposure. The phenomena and indicators of ferroptosis were then determined and verified by ferroptosis inhibitor ferrostatin-1 (Fer-1)

and it was also found that nuclear factor erythroid 2-related factor 2 (Nrf2) played an important role in spermatogenic cell ferroptosis induced by PS-NPs. Finally

it was confirmed in vivo that this mechanism of Nrf2 played a protective role in PS-NPs-induced male reproductive toxicity. This study demonstrated that PS-NPs induce male reproductive dysfunction in mice by causing spermatogenic cell ferroptosis dependent on Nrf2.

关键词

聚苯乙烯纳米塑料（PS-NPs）雄性生殖毒性铁死亡核因子e2相关因子2（Nrf2）

Keywords

Polystyrene nanoplastics (PS-NPs)Reproductive toxicityFerroptosisNuclear factor erythroid 2-related factor 2 (Nrf2)

references

Amereh F, Babaei M, Eslami A, et al., 2020. The emerging risk of exposure to nano(micro)plastics on endocrine disturbance and reproductive toxicity: from a hypothetical scenario to a global public health challenge. Environ Pollut, 261:114158. https://doi.org/10.1016/j.envpol.2020.114158https://doi.org/10.1016/j.envpol.2020.114158

Archibong AE, Rideout ML, Harris KJ, et al., 2018. Oxidative stress in reproductive toxicology. Curr Opin Toxicol, 7:95-101. https://doi.org/10.1016/j.cotox.2017.10.004https://doi.org/10.1016/j.cotox.2017.10.004

Bisht S, Faiq M, Tolahunase M, et al., 2017. Oxidative stress and male infertility. Nat Rev Urol, 14(8):470-485. https://doi.org/10.1038/nrurol.2017.69https://doi.org/10.1038/nrurol.2017.69

Chen X, Comish PB, Tang DL, et al., 2021. Characteristics and biomarkers of ferroptosis. Front Cell Dev Biol, 9:637162. https://doi.org/10.3389/fcell.2021.637162https://doi.org/10.3389/fcell.2021.637162

Cortés C, Domenech J, Salazar M, et al., 2020. Nanoplastics as a potential environmental health factor: effects of polystyrene nanoparticles on human intestinal epithelial Caco-2 cells. Environ Sci Nano, 7(1):272-285. https://doi.org/10.1039/C9EN00523Dhttps://doi.org/10.1039/C9EN00523D

Deng YF, Zhang Y, Lemos B, et al., 2017. Tissue accumulation of microplastics in mice and biomarker responses suggest widespread health risks of exposure. Sci Rep, 7:46687. https://doi.org/10.1038/srep46687https://doi.org/10.1038/srep46687

Dixon SJ, Lemberg KM, Lamprecht MR, et al., 2012. Ferroptosis: an iron-dependent form of nonapoptotic cell death. Cell, 149(5):1060-1072. https://doi.org/10.1016/j.cell.2012.03.042https://doi.org/10.1016/j.cell.2012.03.042

Dodson M, Castro-Portuguez R, Zhang DD, 2019. NRF2 plays a critical role in mitigating lipid peroxidation and ferroptosis. Redox Biol, 23:101107. https://doi.org/10.1016/j.redox.2019.101107https://doi.org/10.1016/j.redox.2019.101107

Du FN, Cai HW, Zhang Q, et al., 2020. Microplastics in take-out food containers. J Hazard Mater, 399:122969. https://doi.org/10.1016/j.jhazmat.2020.122969https://doi.org/10.1016/j.jhazmat.2020.122969

Du X, Zhang JJ, Liu L, et al., 2022. A novel anticancer property of Lycium barbarum polysaccharide in triggering ferroptosis of breast cancer cells. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 23(4):286-299. https://doi.org/10.1631/jzus.B2100748https://doi.org/10.1631/jzus.B2100748

Fendall LS, Sewell MA, 2009. Contributing to marine pollution by washing your face: microplastics in facial cleansers. Mar Pollut Bull, 58(8):1225-1228. https://doi.org/10.1016/j.marpolbul.2009.04.025https://doi.org/10.1016/j.marpolbul.2009.04.025

Fu XF, Liu L, Han H, et al., 2023. Integrated fecal microbiome and metabolome analysis explore the link between polystyrene nanoplastics exposure and male reproductive toxicity in mice. Environ Toxicol, 38(6):1277-1291. https://doi.org/10.1002/tox.23763https://doi.org/10.1002/tox.23763

Gasperi J, Wright SL, Dris R, et al., 2018. Microplastics in air: are we breathing it in? Curr Opin Environ Sci Health, 1:1-5. https://doi.org/10.1016/j.coesh.2017.10.002https://doi.org/10.1016/j.coesh.2017.10.002

Harada N, Kanayama M, Maruyama A, et al., 2011. Nrf2 regulates ferroportin 1-mediated iron efflux and counteracts lipopolysaccharide-induced ferroportin 1 mRNA suppression in macrophages. Arch Biochem Biophys, 508(1):‍101-109. https://doi.org/10.1016/j.abb.2011.02.001https://doi.org/10.1016/j.abb.2011.02.001

Hassannia B, Vandenabeele P, Berghe TV, 2019. Targeting ferroptosis to iron out cancer. Cancer Cell, 35(6):830-849. https://doi.org/10.1016/j.ccell.2019.04.002https://doi.org/10.1016/j.ccell.2019.04.002

Hernandez LM, Xu EG, Larsson HCE, et al., 2019. Plastic teabags release billions of microparticles and nanoparticles into tea. Environ Sci Technol, 53(21):12300-12310. https://doi.org/10.1021/acs.est.9b02540https://doi.org/10.1021/acs.est.9b02540

Huang C, Li BS, Xu KR, et al., 2017. Decline in semen quality among 30,636 young Chinese men from 2001 to 2015. Fertil Steril, 107(1):83-88.e2. https://doi.org/10.1016/j.fertnstert.2016.09.035https://doi.org/10.1016/j.fertnstert.2016.09.035

Imai H, Hakkaku N, Iwamoto R, et al., 2009. Depletion of selenoprotein GPx4 in spermatocytes causes male infertility in mice. J Biol Chem, 284(47):32522-32532. https://doi.org/10.1074/jbc.M109.016139https://doi.org/10.1074/jbc.M109.016139

Jiang L, Wang JM, Wang K, et al., 2021. RNF217 regulates iron homeostasis through its E3 ubiquitin ligase activity by modulating ferroportin degradation. Blood, 138(8):689-705. https://doi.org/10.1182/blood.2020008986https://doi.org/10.1182/blood.2020008986

Jin HB, Ma T, Sha XX, et al., 2021. Polystyrene microplastics induced male reproductive toxicity in mice. J Hazard Mater, 401:123430. https://doi.org/10.1016/j.jhazmat.2020.123430https://doi.org/10.1016/j.jhazmat.2020.123430

Jung BK, Han SW, Park SH, et al., 2020. Neurotoxic potential of polystyrene nanoplastics in primary cells originating from mouse brain. Neurotoxicology, 81:189-196. https://doi.org/10.1016/j.neuro.2020.10.008https://doi.org/10.1016/j.neuro.2020.10.008

Kedzierski M, Lechat B, Sire O, et al., 2020. Microplastic contamination of packaged meat: occurrence and associated risks. Food Packag Shelf Life, 24:100489. https://doi.org/10.1016/j.fpsl.2020.100489https://doi.org/10.1016/j.fpsl.2020.100489

Kerins MJ, Ooi A, 2018. The roles of NRF2 in modulating cellular iron homeostasis. Antioxid Redox Signal, 29(17):1756-1773. https://doi.org/10.1089/ars.2017.7176https://doi.org/10.1089/ars.2017.7176

Khan FR, Catarino AI, Clark NJ, 2022. The ecotoxicological consequences of microplastics and co-contaminants in aquatic organisms: a mini-review. Emerg Top Life Sci, 6(4):339-348. https://doi.org/10.1042/ETLS20220014https://doi.org/10.1042/ETLS20220014

Kim MJ, Yun GJ, Kim SE, 2021. Metabolic regulation of ferroptosis in cancer. Biology, 10(2):83. https://doi.org/10.3390/biology10020083https://doi.org/10.3390/biology10020083

Levine H, Jørgensen N, Martino-Andrade A, et al., 2017. Temporal trends in sperm count: a systematic review and meta-regression analysis. Hum Reprod Update, 23(6):646-659. https://doi.org/10.1093/humupd/dmx022https://doi.org/10.1093/humupd/dmx022

Li SB, He YP, Chen KX, et al., 2021. RSL3 drives ferroptosis through NF‍-‍κB pathway activation and GPX4 depletion in glioblastoma. Oxid Med Cell Longev, 2021:2915019. https://doi.org/10.1155/2021/2915019https://doi.org/10.1155/2021/2915019

Li YM, Liu ZQ, Yang Y, et al., 2021. Effects of nanoplastics on energy metabolism in the oriental river prawn (Macrobrachium nipponense). Environ Pollut, 268:115890. https://doi.org/10.1016/j.envpol.2020.115890https://doi.org/10.1016/j.envpol.2020.115890

Liu ZX, Lv XY, Yang BW, et al., 2021. Tetrachlorobenzoquinone exposure triggers ferroptosis contributing to its neurotoxicity. Chemosphere, 264:128413. https://doi.org/10.1016/j.chemosphere.2020.128413https://doi.org/10.1016/j.chemosphere.2020.128413

Lu J, Zhao YN, Liu MT, et al., 2021. Toward improved human health: Nrf2 plays a critical role in regulating ferroptosis. Food Funct, 12(20):9583-9606. https://doi.org/10.1039/d1fo01036khttps://doi.org/10.1039/d1fo01036k

Lusher A, Hollman P, Mendoza-Hill J, 2017. Microplastics in Fisheries and Aquaculture: Status of Knowledge on Their Occurrence and Implications for Aquatic Organisms and Food Safety. FAO Fisheries and Aquaculture Technical Paper No. 615, Food and Agriculture Organization of the United Nations, Madrid.

Meng XM, Zhang JW, Wang WJ, et al., 2022. Effects of nano- and microplastics on kidney: physicochemical properties, bioaccumulation, oxidative stress and immunoreaction. Chemosphere, 288(Pt 3):132631. https://doi.org/10.1016/j.chemosphere.2021.132631https://doi.org/10.1016/j.chemosphere.2021.132631

Miotto G, Rossetto M, di Paolo ML, et al., 2020. Insight into the mechanism of ferroptosis inhibition by ferrostatin-1. Redox Biol, 28:101328. https://doi.org/10.1016/j.redox.2019.101328https://doi.org/10.1016/j.redox.2019.101328

Nair AB, Jacob S, 2016. A simple practice guide for dose conversion between animals and human. J Basic Clin Pharm, 7(2):27-31. https://doi.org/10.4103/0976-0105.177703https://doi.org/10.4103/0976-0105.177703

Oßmann BE, Sarau G, Holtmannspötter H, et al., 2018. Small-sized microplastics and pigmented particles in bottled mineral water. Water Res, 141:307-316. https://doi.org/10.1016/j.watres.2018.05.027https://doi.org/10.1016/j.watres.2018.05.027

Park MW, Cha HW, Kim J, et al., 2021. NOX4 promotes ferroptosis of astrocytes by oxidative stress-induced lipid peroxidation via the impairment of mitochondrial metabolism in Alzheimer’s diseases. Redox Biol, 41:101947. https://doi.org/10.1016/j.redox.2021.101947https://doi.org/10.1016/j.redox.2021.101947

Ragusa A, Svelato A, Santacroce C, et al., 2021. Plasticenta: first evidence of microplastics in human placenta. Environ Int, 146:106274. https://doi.org/10.1016/j.envint.2020.106274https://doi.org/10.1016/j.envint.2020.106274

Riedelberger M, Penninger P, Tscherner M, et al., 2020. Type I interferon response dysregulates host iron homeostasis and enhances Candida glabrata infection. Cell Host Microbe, 27(3):454-466.e8. https://doi.org/10.1016/j.chom.2020.01.023https://doi.org/10.1016/j.chom.2020.01.023

Rist S, Baun A, Hartmann NB, 2017. Ingestion of micro- and nanoplastics in Daphnia magna‍ ‒‍ quantification of body burdens and assessment of feeding rates and reproduction. Environ Pollut, 228:398-407. https://doi.org/10.1016/j.envpol.2017.05.048https://doi.org/10.1016/j.envpol.2017.05.048

Schulte RT, Ohl DA, Sigman M, et al., 2010. Sperm DNA damage in male infertility: etiologies, assays, and outcomes. J Assist Reprod Genet, 27(1):3-12. https://doi.org/10.1007/s10815-009-9359-xhttps://doi.org/10.1007/s10815-009-9359-x

Shukla S, Khan R, Saxena A, et al., 2022. Microplastics from face masks: a potential hazard post Covid-19 pandemic. Chemosphere, 302:134805. https://doi.org/10.1016/j.chemosphere.2022.134805https://doi.org/10.1016/j.chemosphere.2022.134805

Song XH, Long DX, 2020. Nrf2 and ferroptosis: a new research direction for neurodegenerative diseases. Front Neurosci, 14:267. https://doi.org/10.3389/fnins.2020.00267https://doi.org/10.3389/fnins.2020.00267

Stockwell BR, Jiang XJ, Gu W, 2020. Emerging mechanisms and disease relevance of ferroptosis. Trends Cell Biol, 30(6):478-490. https://doi.org/10.1016/j.tcb.2020.02.009https://doi.org/10.1016/j.tcb.2020.02.009

Sun XF, Ou ZH, Chen RC, et al., 2016. Activation of the p62-Keap1-NRF2 pathway protects against ferroptosis in hepatocellular carcinoma cells. Hepatology, 63(1):173-184. https://doi.org/10.1002/hep.28251https://doi.org/10.1002/hep.28251

Tang DL, Chen X, Kang R, et al., 2021. Ferroptosis: molecular mechanisms and health implications. Cell Res, 31(2):107-125. https://doi.org/10.1038/s41422-020-00441-1https://doi.org/10.1038/s41422-020-00441-1

Tao SS, Liu PF, Luo G, et al., 2017. p97 negatively regulates NRF2 by extracting ubiquitylated NRF2 from the KEAP1-CUL3 E3 complex. Mol Cell Biol, 37(8):e00660-16. https://doi.org/10.1128/MCB.00660-16https://doi.org/10.1128/MCB.00660-16

Ursini F, Maiorino M, 2020. Lipid peroxidation and ferroptosis: the role of GSH and GPx4. Free Radic Biol Med, 152:175-185. https://doi.org/10.1016/j.freeradbiomed.2020.02.027https://doi.org/10.1016/j.freeradbiomed.2020.02.027

Wang LW, Wu WM, Bolan NS, et al., 2021. Environmental fate, toxicity and risk management strategies of nanoplastics in the environment: current status and future perspectives. J Hazard Mater, 401:123415. https://doi.org/10.1016/j.jhazmat.2020.123415https://doi.org/10.1016/j.jhazmat.2020.123415

Wang Z, An CJ, Chen XJ, et al., 2021. Disposable masks release microplastics to the aqueous environment with exacerbation by natural weathering. J Hazard Mater, 417:126036. https://doi.org/10.1016/j.jhazmat.2021.126036https://doi.org/10.1016/j.jhazmat.2021.126036

Xu DH, Ma YH, Han XD, et al., 2021. Systematic toxicity evaluation of polystyrene nanoplastics on mice and molecular mechanism investigation about their internalization into Caco-2 cells. J Hazard Mater, 417:126092. https://doi.org/10.1016/j.jhazmat.2021.126092https://doi.org/10.1016/j.jhazmat.2021.126092

Xu MK, Halimu G, Zhang QR, et al., 2019. Internalization and toxicity: a preliminary study of effects of nanoplastic particles on human lung epithelial cell. Sci Total Environ, 694:133794. https://doi.org/10.1016/j.scitotenv.2019.133794https://doi.org/10.1016/j.scitotenv.2019.133794

Yee MSL, Hii LW, Looi CK, et al., 2021. Impact of microplastics and nanoplastics on human health. Nanomaterials, 11(2):496. https://doi.org/10.3390/nano11020496https://doi.org/10.3390/nano11020496

Yin K, Wang Y, Zhao HJ, et al., 2021. A comparative review of microplastics and nanoplastics: toxicity hazards on digestive, reproductive and nervous system. Sci Total Environ, 774:145758. https://doi.org/10.1016/j.scitotenv.2021.145758https://doi.org/10.1016/j.scitotenv.2021.145758

Yin LS, Wen XF, Huang DL, et al., 2021. Interactions between microplastics/nanoplastics and vascular plants. Environ Pollut, 290:117999. https://doi.org/10.1016/j.envpol.2021.117999https://doi.org/10.1016/j.envpol.2021.117999

Yong CQY, Valiyaveettil S, Tang BL, 2020. Toxicity of microplastics and nanoplastics in mammalian systems. Int J Environ Res Public Health, 17(5):1509. https://doi.org/10.3390/ijerph17051509https://doi.org/10.3390/ijerph17051509

Yosri N, Khalifa SAM, Guo ZM, et al., 2021. Marine organisms: pioneer natural sources of polysaccharides/proteins for green synthesis of nanoparticles and their potential applications. Int J Biol Macromol, 193:1767-1798. https://doi.org/10.1016/j.ijbiomac.2021.10.229https://doi.org/10.1016/j.ijbiomac.2021.10.229

Zegers-Hochschild F, Adamson GD, Dyer S, et al, 2017. The international glossary on infertility and fertility care, 2017. Fertil Steril, 108(3):393-406. https://doi.org/10.1016/j.fertnstert.2017.06.005https://doi.org/10.1016/j.fertnstert.2017.06.005

More>

Views

Downloads

CSCD

Tools

Publicity Resources

Targeting ferroptosis and ferritinophagy: new targets for cardiovascular diseases

Novel perspective in transplantation therapy of mesenchymal stem cells: targeting the ferroptosis pathway

Iron metabolism, ferroptosis, and lncRNA in cancer: knowns and unknowns

A novel anticancer property of Lycium barbarum polysaccharide in triggering ferroptosis of breast cancer cells

Exosomal let-7a-5p derived from human umbilical cord mesenchymal stem cells alleviates coxsackievirus B3-induced cardiomyocytes ferroptosis via the SMAD2/ZFP36 signal axis

Related Author

No data

Related Institution

Henan Engineering Research Center for Application & Translation of Precision Clinical Pharmacy, Zhengzhou University

Clinical Systems Biology Research Laboratories, Translational Medicine Center, the First Affiliated Hospital of Zhengzhou University

State Key Laboratory for Artificial Microstructures and Mesoscopic Physics, School of Physics, Peking University

School of Laboratory Medicine, Xinxiang Medical University

Quality Management Department, Henan No. 3 Provincial People’s Hospital

Address：Journal of Zhejiang University-SCIENCE, 38 Zheda Road, Hangzhou, China Postal code：310027
Tel：+86-571-87952783 Email：cjzhang@zju.edu.cn
Technical support is provided by Beijing Founder electronics co., LTD 京ICP备09064830号-19 京公网安备11010802024621
It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.

⁰