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WDR74 facilitates TGF-β/Smad pathway activation to promote M2 macrophage polarization and diabetic foot ulcer wound healing in mice

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Abstract

Diabetic foot ulcer (DFU) is a devastating component of diabetes progression, leading to decreased quality of life and increased mortality in diabetic patients. The underlying mechanism of DFU is not completely understood. Hence, this study aims to elucidate the mechanism involved in wound healing in mouse models of DFU. Gain- and loss-of-function studies were performed to study the roles that WDR74 and TGF-β play in mouse models of DFU and primary bone marrow–derived mouse macrophages. M1 and M2 macrophage phenotypic markers, extracellular matrix (ECM) components, and angiogenic makers were determined by RT-qPCR and/or Western blot analysis. Localization of these proteins was determined by immunofluorescence staining and/or immunohistochemistry. Interaction between WDR74 with Smad2/3 in macrophages was detected by co-immunoprecipitation. We found that WDR74 and M2 macrophages were decreased in wound tissues from DFU mice. TGF-β/Smad pathway activation increased the expression of M2 macrophage markers (arginase-1 and YM1), IL-4, while decreased expression of M1 macrophage marker (iNOS). TGF-β/Smad pathway activation also increased the production of ECM and promoted the wound closure in diabetic mice. We also noticed that WDR74 overexpression increased Smad2/3 phosphorylation, elevated the population of M2 macrophage and ECM production, and alleviated DFU. LY2109761 treatment normalized effects of TGF-β or WDR74 overexpression. In conclusion, WDR74 promoted M2 macrophage polarization, leading to improved DFU in mice, through activation of the TGF-β/Smad pathway.

Graphical abstract

Graphical Headlights

1. WDR74 promotes M2 macrophage polarization and ECM production.

2. WDR74 activates the TGF-β/Smad signaling pathway.

3. TGF-β/Smad activation promotes M2 macrophage polarization in murine DFU.

4. WDR74 enhances wound healing in murine DFU.

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Data availability

All datasets generated for this study are included in the article/Supplementary Material.

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Funding

The work was supported by grants from the National Natural Science Foundation of China (Nos. 81970676 and NO.82170834), the Science and Technology Development Fund of Macau (No. 0055/2019/AMJ), the Office of Science & Technology and Talent Work of Luzhou (No. 2019-JYJ-63), and the Innovation Seedling Project of Sichuan Provincial Department of Science and Technology (No. 2022095).

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Author notes

  1. Kang Geng and Xiumei Ma contributed equally to this work.

Authors and Affiliations

  1. Faculty of Chinese Medicine, State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Avenida Wai Long, Taipa, Macao

    Kang Geng, Xiumei Ma, Weiming Wang, Yong Xu & Youhua Xu

  2. Department of Endocrinology and Metabolism, Metabolic Vascular Disease Key Laboratory of Sichuan Province, Sichuan Clinical Research Center for Nephropathy, Luzhou, Sichuan, People’s Republic of China

    Kang Geng, Xiumei Ma, Zongzhe Jiang, Wei Huang & Yong Xu

  3. Department of Plastic and Burn Surgery, National Key Clinical Construction Specialty, The Affiliated Hospital of Southwest Medical University, Luzhou, Sichuan, People’s Republic of China

    Kang Geng

  4. Endocrinology Department, The Second People’s Hospital of Yibin, West China Yibin Hospital, Sichuan University, Yibin, Sichuan, People’s Republic of China

    Junling Gu

  5. Department of General Surgery (Vascular Surgery), The Affiliated Hospital of Southwest Medical University, Luzhou, Sichuan, People’s Republic of China

    Weiming Wang

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Contributions

Youhua Xu provided the scope of the research and conceived the manuscript structure. Kang Geng and Xiumei Ma oversaw the writing of the manuscript and drafted the final version of the manuscript. Zongzhe Jiang, Huang Wei, Junling Gu, and Weiming Wang were involved in data collection. Xiumei Ma and Yong Xu performed the statistical analysis and preparation of the Figures. Kang Geng and Youhua Xu drafted the paper. All authors read and approved the final manuscript.

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Correspondence to Yong Xu or Youhua Xu.

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All mice were maintained in the pathogen-free animal facility and all animal studies were approved by the Institutional Animal Care and Use Committee at The Affiliated Hospital of Southwest Medical University.

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Geng, K., Ma, X., Jiang, Z. et al. WDR74 facilitates TGF-β/Smad pathway activation to promote M2 macrophage polarization and diabetic foot ulcer wound healing in mice. Cell Biol Toxicol 39, 1577–1591 (2023). https://doi.org/10.1007/s10565-022-09748-8

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