Inhibition of ferroptosis attenuates oligospermia in male Nrf2 knockout mice

Ping Han; Xia Wang; Tianqiu Zhou; Jinmei Cheng; Chengniu Wang; Fei Sun; Xi Zhao

doi:10.1016/j.freeradbiomed.2022.10.314

Inhibition of ferroptosis attenuates oligospermia in male Nrf2 knockout mice

Free Radic Biol Med. 2022 Nov 20;193(Pt 1):421-429. doi: 10.1016/j.freeradbiomed.2022.10.314. Epub 2022 Oct 27.

Authors

Ping Han¹, Xia Wang², Tianqiu Zhou³, Jinmei Cheng¹, Chengniu Wang¹, Fei Sun⁴, Xi Zhao¹

Affiliations

¹ Institute of Reproductive Medicine, Medical School, Nantong University, Nantong, 226001, Jiangsu Province, China.
² Center for Reproductive Medicine, Affiliated Hospital of Nantong University, Nantong, 226001, Jiangsu Province, China.
³ Department of Ophthalmology, Affiliated Hospital of Nantong University, Nantong, 226001, Jiangsu Province, China.
⁴ Institute of Reproductive Medicine, Medical School, Nantong University, Nantong, 226001, Jiangsu Province, China. Electronic address: sunfei@ntu.edu.cn.

PMID: 36309297
DOI: 10.1016/j.freeradbiomed.2022.10.314

Abstract

Nuclear factor-E2-related factor 2 (Nrf2) expression in sperm decreases in some oligospermia patients. However, the mechanism of reduced Nrf2 expression in sperm of oligospermia men is not elucidated. In the present study, our clinical trial results showed that Nrf2 and glutathione peroxidase 4 (GPX4) protein expressions in sperm of oligospermia men significantly decreased than those of healthy men. In animal experiments, mice were randomly divided into 3 groups: wild type (WT), Nrf2 knockout (Nrf2^-/-) and Nrf2^-/- + ferroptosis inhibitor (Fer-1) groups. Fer-1 was intraperitoneally injected in Nrf2^-/- mice for 4 weeks. The results showed that male Nrf2^-/- mice displayed decreased sperm concentration and motility, and significantly lower fertility. Compared with WT mice, malondialdehyde (MDA) content and prostaglandin-endoperoxide synthase 2 (Ptgs2) mRNA expression increased, but nicotinamide adenine dinucleotide phosphate oxidase (NADPH) content decreased in the testes of Nrf2^-/- mice, which were biomarkers of ferroptosis. Furthermore, treatment with Fer-1 in Nrf2^-/- mice reversed the decreased sperm concentration and motility. Meanwhile, histology showed that spermatogenic cells obviously decreased, and vacuolization formed in the testes of Nrf2^-/- mice, which were reversed by Fer-1 treatment. Additionally, compared with WT mice, GPX4, solute carrier family 7 member 11 (SLC7A11), glutamate-cysteine ligase, catalytic subunit (Gclc), glutamate-cysteine ligase, modifier subunit (Gclm) and ferroportin 1 (FPN1) mRNA and protein expressions significantly decreased, but transferrin receptor 1 (TfR1) and divalent metal transporter 1 (DMT1) mRNA and protein expressions increased in testicular tissues in Nrf2^-/- mice. After treatment with Fer-1, only Gclc and Gclm mRNA and protein expressions increased. Taken together, our data suggested that deletion of Nrf2 leads to downregulation of GPX4 and regulation of other ferroptosis-related genes, resulting in ferroptosis occurrence in spermatogenic cells and ultimately oligospermia.

Keywords: Ferroptosis; GPX4; Nrf2; Oligospermia; SLC7A11; TfR1.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cyclooxygenase 2
Ferroptosis* / genetics
Glutamate-Cysteine Ligase / genetics
Humans
Male
Mice
Mice, Knockout
NF-E2-Related Factor 2 / metabolism
Oligospermia* / genetics
Phospholipid Hydroperoxide Glutathione Peroxidase
RNA, Messenger
Semen / metabolism

Substances

NF-E2-Related Factor 2
Glutamate-Cysteine Ligase
Phospholipid Hydroperoxide Glutathione Peroxidase
Cyclooxygenase 2
RNA, Messenger