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4-OI ameliorates bleomycin-induced pulmonary fibrosis by activating Nrf2 and suppressing macrophage-mediated epithelial-mesenchymal transition

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Abstract

Objectives

Pulmonary fibrosis (PF) is a chronic and refractory interstitial lung disease with limited therapeutic options. 4-octyl itaconate (4-OI), a cell-permeable derivative of itaconate, has been shown to have anti-oxidative and anti-inflammatory properties. However, the effect and the underlying mechanism of 4-OI on PF are still unknown.

Methods

WT or Nrf2 knockout (Nrf2−/−) mice were intratracheally injected with bleomycin (BLM) to establish PF model and then treated with 4-OI. The mechanism study was performed by using RAW264.7 cells, primary macrophages, and conditional medium-cultured MLE-12 cells.

Results

4-OI significantly alleviated BLM-induced PF and EMT process. Mechanism studies have found that 4-OI can not only directly inhibit EMT process, but also can reduce the production of TGF-β1 by restraining macrophage M2 polarization, which in turn inhibits EMT process. Moreover, the effect of 4-OI on PF and EMT depends on Nrf2.

Conclusion

4-OI ameliorates BLM-induced PF in an Nrf2-dependent manner, and its role in alleviating PF is partly due to the direct inhibition on EMT, and partly through indirect inhibition of M2-mediated EMT. These findings suggested that 4-OI has great clinical potential to develop as a new anti-fibrotic agent for PF therapy.

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Data availability statement

The authors will supply the relevant data in response to reasonable requests.

Abbreviations

PF:

Pulmonary fibrosis

BLM:

Bleomycin

4-OI:

4-Octyl itaconate

EMT:

Epithelial-mesenchymal transition

AT II:

Alveolar epithelial type II

Nrf2:

Nuclear factor E2-related factor 2

E-cad:

E-cadherin

α-SMA:

α-Smooth muscle actin

Fn:

Fibronectin

MMP2:

Matrix metalloproteinase 2

MTA1:

Metastasis-associated gene 1

M1:

Classically activated macrophages

M2:

Alternatively activated macrophages

Arg-1:

Arginase-1

Ym1:

Chitinase-like protein 3

Fizz1:

Resistin-like molecule α1

IL-4:

Interleukin-4

IL-13:

Interleukin-13

BALF:

Bronchoalveolar lavage fluid

PBS:

Phosphate-buffered saline

M-csf:

Macrophage-colony-stimulating factor

Col1a1:

Collagen alpha1

Keap1:

Kelch-like ECH-associated protein 1

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Acknowledgements

This study was supported by the National Natural Science Foundation of China (82100018); Natural Science Foundation of Jiangsu Province (BK20200602); China Postdoctoral Science Foundation (2020M671347); National Natural Science Foundation of China (82171704).

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Authors and Affiliations

  1. Wuxi School of Medicine, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, Jiangsu, People’s Republic of China

    Ya-xian Wu, Ya-ru Zhang, Feng-juan Jiang, Shuai He, Yan-li Zhang, Dan Chen, Ying Tong, Yun-juan Nie & Qing-feng Pang

  2. Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, Jiangsu, People’s Republic of China

    Ya-xian Wu

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Contributions

YXW and QFP designed the study and drafted the paper; YJN participated in the design and helped draft the paper; YXW, YRZ and FJJ performed the experiments; SH and YLZ participated in the animal experiments; DC and YT performed the data analysis; YXW and YJN were involved in the discussion of the experiments. All authors read and approved the final paper.

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Correspondence to Yun-juan Nie or Qing-feng Pang.

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Wu, Yx., Zhang, Yr., Jiang, Fj. et al. 4-OI ameliorates bleomycin-induced pulmonary fibrosis by activating Nrf2 and suppressing macrophage-mediated epithelial-mesenchymal transition. Inflamm. Res. 72, 1133–1145 (2023). https://doi.org/10.1007/s00011-023-01733-z

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