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NRF2 is essential for iron-overload stimulated osteoclast differentiation through regulation of redox and iron homeostasis

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Abstract

Iron overload enhances osteoclastic bone resorption and induces osteoporosis. Excess iron is highly toxic. The modulation of redox and iron homeostasis is critical for osteoclast differentiation under iron-overload condition. Nuclear factor erythroid 2-related factor 2 (NRF2) is a transcription factor that regulates the cellular defense against oxidative stress and iron overload through the expression of genes involved in anti-oxidative processes and iron metabolism. Our studies demonstrated that NRF2 activation was suppressed during osteoclast differentiation. Under iron-overload condition, NRF2 and its mediated antioxidant and iron metabolism genes were activated by reactive oxygen species (ROS), which enhanced antioxidant capability. NRF2 mediated the upregulation of iron exporter ferroportin 1 (FPN1) and iron storage protein ferritin, contributing to decreased levels of intracellular iron. Nfe2l2 knockout induced oxidative stress and promoted osteoclast differentiation under normal condition, but induced ferroptosis under iron-overload condition. Nfe2l2 knockout alleviated iron overload induced bone loss by inhibiting osteoclast differentiation. Our results suggest that NRF2 activation is essential for osteoclast differentiation by enhancing antioxidant capability and reducing intracellular iron under iron-overload condition. Targeting NRF2 to induce ferroptosis could be a potential therapy for the treatment of iron-overload induced osteoporosis.

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Data availability

The datasets presented in the current study are available from the corresponding author on reasonable request.

Abbreviations

4-HNE:

4-hydroxynonenal

AREs:

Antioxidant response elements

BMD:

Bone mineral density

BS:

Bone surface

BV:

Bone volume

CAT:

Catalase

DFO:

Deferoxamine

FTH:

Ferritin heavy chain

FTL:

Ferritin light chain

GCLC:

Glutamate-cysteine ligase catalytic subunit

GSH-Px:

Glutathione peroxidase

HO-1:

Heme oxygenase 1

Keap1:

Kelch-like ECH-associated protein 1

LIP:

Labile iron pool

NAC:

N-acetylcysteine

NQO-1:

NAD(P)H dehydrogenase quinone 1

MDA:

Malondialdehyde

NRF2:

Nuclear factor erythroid 2-related factor 2

FPN1:

Ferroportin 1

RANKL:

Receptor activator of nuclear factor kappa-Β ligand

ROS:

Reactive oxygen species

Tb.Th.:

Trabecular thickness

Tb.Sp.:

Trabecular separation

Tb.N.:

Trabecular number

TfR1:

Transferrin receptor 1

TV:

Tissue volume

TRAP:

Tartrate-resistant acid phosphatase

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Funding

This work was supported by Guizhou Provincial Science and Technology Projects (Qian Ke He Ji Chu-ZK[2021]-Yi Ban 398), the National Natural Science Foundation of China (82060168, 82360176), and the Guizhou Education Department Youth Science and Technology Talents Growth Project (Qian Jiao He KY Zi [2021]199).

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Author notes

  1. Jian Zhang and Lingyan Zhang contributed equally to this work.

Authors and Affiliations

  1. Institute of Laboratory Animal Science, Guizhou University of Traditional Chinese Medicine, Huaxi University Town, Guiyang, 550025, Guizhou, China

    Jian Zhang, Lingyan Zhang, Gang Yao, Hai Zhao & Shuguang Wu

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  1. Jian Zhang
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  2. Lingyan Zhang
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  3. Gang Yao
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  4. Hai Zhao
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Contributions

Jian Zhang and Lingyan Zhang contributed to the study conception and design. Material preparation, data collection and analysis were performed by Gang Yao, Hai Zhao, and Shuguang Wu. The manuscript and figures was prepared by Jian Zhang. All authors reviewed the manuscirpt.

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Correspondence to Jian Zhang.

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Ethics approval

The animal study was approved by the Animal Ethics and Welfare Committee at Guizhou University of Traditional Chinese Medicine. All animal experiments complied with the National Research Council’s Guide for the Care and Use of Laboratory Animals.

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Supplementary information

ESM 1

Figure S1 (A) The protein expression of Nrf2 in Nfe2l2 knockout RAW264.7 cells. (B) The sequencing results of Nfe2l2 gene in WT and KO RAW264.7 cells. Figure S2 (A) The protein expression of Nrf2 in Nfe2l2 knockout C57BL/6J mice. (B) The sequencing results of Nfe2l2 gene in WT and KO C57BL/6J mice. (DOCX 161 kb)

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Zhang, J., Zhang, L., Yao, G. et al. NRF2 is essential for iron-overload stimulated osteoclast differentiation through regulation of redox and iron homeostasis. Cell Biol Toxicol 39, 3305–3321 (2023). https://doi.org/10.1007/s10565-023-09834-5

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