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Macrophage pyroptosis promotes synovial fibrosis through the HMGB1/TGF- β1 axis: an in vivo and in vitro study

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Abstract

Macrophages and fibroblasts are the main effector cells in synovial tissue in the knee joint. Our previous studies showed that there was synovial macrophage pyroptosis in knee osteoarthritis (KOA) and that inhibiting this pyroptosis could alleviate synovial fibrosis. In the present study, we aimed to elucidate the mechanism by which macrophage pyroptosis affects synovial fibrosis. We established an LPS/ATP-induced model in macrophages that mimicked the inflammatory environment of KOA and induced macrophage pyroptosis. The TGF-β1, SMAD3, and P-SMAD3, and the synovial fibrosis markers (Collagen I, TIMP1, Vimentin, and TGF-β1) were significantly decreased after fibroblasts were cultured with RAGE inhibitors and SMAD3 inhibitors. Moreover, ELISA and immunofluorescence analysis showed that macrophage pyroptosis induced the release of IL-1β, IL-18, and HMGB1 and caused the translocation of HMGB1 from the fibroblast nucleus to the cell membrane, where it could bind with RAGE. Subsequently, in the synovial tissue of KOA model rats, we observed that inhibiting HMGB1, RAGE, and SMAD3 could alleviate the expression of synovial fibrosis markers (Collagen I, TIMP1, Vimentin, and TGF-β1) at both the mRNA and protein levels. Besides, HE and Sirius Red staining were used to observe the transverse diameter of the right knee. In conclusion, macrophage pyroptosis induced IL-1β, IL-18, and HMGB1, which could be caused HMGB1 to translocate from the fibroblast nucleus and bind with RAGE, activating the TGF-β1/SMAD3 signaling pathway and affecting synovial fibrosis.

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Data availability

All results and data are kept in the section for Departments of Orthopedics, the Affiliated Hospital of the Nan**g University of Chinese Medicine, Nan**g, China. These will be made available from the corresponding author on reasonable request.

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Acknowledgements

The authors wish to express their gratitude to all staff in the Medical Research Center of the First College of Clinical Medicine, especially all the teachers in the Tang ZhongYing Science and Technology Building, Nan**g University of Chinese Medicine, Nan**g, China.

Funding

The current work was supported by the National Natural Science Foundation of China (82004391 and 82205143), the third batch of Peak Academic Talents of Jiangsu Hospital of Traditional Chinese Medicine (y2021rc20), the Jiangsu Provincial Medical Key Discipline (Laboratory) Cultivation Unit (js2252), and the Excellent Young Doctor Program of Jiangsu Hospital of Chinese Medicine (2023QB0122).

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  1. Peng Wu and Taiyang Liao contributed equally to this work.

Authors and Affiliations

  1. Affiliated Hospital of Nan**g University of Chinese Medicine, Nan**g, 210029, China

    Peng Wu, Taiyang Liao, Zhenyuan Ma, Yibao Wei, Songjiang Yin, Zhengquan Huang & Jun Mao

  2. Jiangsu Province Hospital of Chinese Medicine, Nan**g, 210029, China

    Peng Wu, Taiyang Liao, Zhenyuan Ma, Yibao Wei, Songjiang Yin, Zhengquan Huang & Jun Mao

  3. Key Laboratory for Metabolic Diseases in Chinese Medicine, First College of Clinical Medicine, Nan**g University of Chinese Medicine, Nan**g, 210023, China

    Peng Wu, Taiyang Liao, Zhenyuan Ma & Yibao Wei

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  1. Peng Wu
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  2. Taiyang Liao
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Contributions

Peng Wu, Zhengquan Huang, Taiyang Liao, and Jun Mao conceived the study and drafted the manuscript. Zhenyuan Ma, Songjiang Yin, and Yibao Wei designed and performed the animal experiments. Taiyang Liao and Peng Wu performed cell experiments. All the authors have read and agreed to the published version of the manuscript.

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Correspondence to Zhengquan Huang or Jun Mao.

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Ethics approval and consent to participate

This study was permitted by the Animal Ethics Committee of Nan**g University of Chinese Medicine (Nan**g, China) on May 2019 (approval number 201905A002). All experiments were conducted in accordance with the National Institutes of Health guidelines for the care and use of laboratory animals.

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The authors declare no competing interests.

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Wu, P., Liao, T., Ma, Z. et al. Macrophage pyroptosis promotes synovial fibrosis through the HMGB1/TGF- β1 axis: an in vivo and in vitro study. In Vitro Cell.Dev.Biol.-Animal 59, 289–299 (2023). https://doi.org/10.1007/s11626-023-00769-z

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