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The immunomodulatory effects of metformin in LPS-induced macrophages: an in vitro study

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Abstract

Objective

The study aimed to explore the immunomodulatory effects of clinically relevant concentrations of metformin on macrophages during sepsis, which is characterized by an initial hyperinflammatory phase followed by a period of immunosuppression. 

Methods

We employed the RAW 264.7 mouse macrophage cell line as an in vitro model to induce inflammatory responses and immune suppression through primary and secondary stimulation by lipopolysaccharide (LPS). The cells were exposed to clinically relevant concentrations of metformin, and their responses were gauged through cytotoxicity assays, enzyme-linked immunosorbent assay for cytokine quantification, and assessments of intracellular reactive oxygen species (ROS) production. Moreover, to probe the role of AMPK in mediating the effects of metformin, we conducted an AMP-activated protein kinase (AMPK) activity assay and knocked down AMPK using siRNA. 

Results

Our study revealed that clinically relevant concentrations of metformin considerably decreased the LPS-induced secretion of tumor necrosis factor-α and interleukin-6, which indicates the suppression of the initial hyperinflammatory response. Furthermore, metformin prevented LPS-induced immunosuppression. Notably, these immunomodulatory effects of metformin were not mediated by the activation of the AMPK pathway, as evidenced by the unaltered AMPK activity and siRNA experiments. The modulation of intracellular ROS levels emerged as the critical mechanism underlying the inhibition of hyperinflammation and impediment of immunosuppression by metformin.

Conclusion

A certain therapeutic dose of metformin inhibited hyperinflammatory responses and alleviated immunosuppression in LPS-induced macrophages through the bidirectional modulation of intracellular ROS generation.

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Data availability

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

The present research was supported by the Guizhou Province Science and Technology Foundation ([2020]1Y403), and the Scientific Research Foundation of Zunyi Medical University (F955 and FZH004).

Funding

Scientific Research Foundation of Zunyi Medical University,F955 and FZH004, F955 and FZH004, Guizhou Province Science and Technology Foundation, [2020]1Y403.

Author information

Authors and Affiliations

  1. Department of Immunology, Zhuhai Campus of Zunyi Medical University, Zhuhai, China

    Zhiyong Wang & Pei Wei

  2. Department of Pharmaceutics, Zhuhai Campus of Zunyi Medical University, Zhuhai, China

    Min Wang

  3. Department of Physiology, Zhuhai Campus of Zunyi Medical University, Zhuhai, China

    Mao Lin

Authors
  1. Zhiyong Wang
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  2. Min Wang
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  3. Mao Lin
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  4. Pei Wei
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Contributions

W. P. designed the experiments. W. Z. and L. M. performed the experimental work. W. M. analyzed the data. W. Z. and L. M. wrote the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Pei Wei.

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The authors declare no conflicts of interest in association with this study.

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Wang, Z., Wang, M., Lin, M. et al. The immunomodulatory effects of metformin in LPS-induced macrophages: an in vitro study. Inflamm. Res. 73, 175–181 (2024). https://doi.org/10.1007/s00011-023-01827-8

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  • DOI: https://doi.org/10.1007/s00011-023-01827-8

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