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3H-1,2-dithiole-3-thione suppresses LPS-induced proinflammatory responses in macrophages: potential involvement of antioxidant induction, NF-κB, and Nrf2

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Abstract

Previously, we reported that 3H-1,2-dithiole-3-thione (D3T), an Nrf2 activator, acted as a potential chemoprotectant against lipopolysaccharide (LPS)-induced mortality in mice. In view of the critical involvement of macrophages in the pathogenesis of LPS-induced endotoxemia, in the present study, we investigated the protective effects of D3T on LPS-induced proinflammatory responses in cultured murine RAW 264.7 macrophage cell line and primary peritoneal macrophages and the potential involvement of antioxidant induction, NF-κB, and Nrf2. We showed that treatment with D3T resulted in increased levels of a series of antioxidants in RAW 264.7 cells in a concentration-dependent manner. These included the reduced form of glutathione, glutathione peroxidase, glutathione reductase, glutathione S-transferase, and NADPH:quinone oxidoreductase 1. Catalase was also potently induced by D3T which, however, did not show a concentration dependency. Concurrent with the ability to induce the above cellular antioxidants, D3T pretreatment of RAW 264.7 cells also led to a concentration-dependent suppression of LPS-induced interleukin-1beta (IL-1β) production and nitric oxide release. LPS-stimulated tumor necrosis factor-alpha (TNF-α) production was also suppressed by D3T, but to a much lesser extent. Using NF-κB reporter gene-expressing RAW 264.7 cells, we further showed that D3T pretreatment also suppressed LPS-induced NF-κB activation. To investigate the potential involvement of Nrf2, a chief regulator of cellular antioxidant genes, we used peritoneal macrophages isolated from Nrf2+/+ and Nrf2−/− mice. Our results showed that D3T pretreatment suppressed LPS-induced proinflammatory responses in Nrf2+/+ macrophages, and this inhibitory effect of D3T was completely lost in Nrf2−/− macrophages. Collectively, the results of the present study demonstrated that D3T acted as a potent suppressor of LPS-induced proinflammatory responses in macrophages. Antioxidant induction, NF-κB suppression, and Nrf2 activation appeared to contribute to the anti-proinflammatory activity of D3T in macrophages.

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Data availability

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

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Acknowledgements

This study was supported in part by a grant from the National Institutes of Health (GM124652). The Berthold LB9505 luminometer was provided by Dr. Michael A. Trush from The Johns Hopkins University (Baltimore, MD).

Funding

This study was supported in part by a Grant (GM124652) from the National Institutes of Health, Bethesda, MD.

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Authors and Affiliations

  1. Campbell University Jerry M. Wallace School of Osteopathic Medicine, Buies Creek, NC, 27506, USA

    Hong Zhu, An Bui & Y. Robert Li

  2. Edward Via College of Osteopathic Medicine, Virginia Tech Corporate Research Center, Blacksburg, VA, 24060, USA

    Arben Santo

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  1. Hong Zhu
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All authors contributed to the study conception, design, performance, data analysis, and writing. All authors read and approved the final manuscript.

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Correspondence to Hong Zhu.

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The authors have no relevant financial or non-financial interest to disclose.

Ethical approval

This work does not directly involve intact animals or human subjects. The experimental protocol for the isolation of the primary peritoneal macrophages from Nrf2+/+ and Nrf2−/− mice (on C57BL/6 background) was approved by the Institutional Animal Care and Use Committee as described in Ref [9], and the cells has been kept in liquid nitrogen ever since and were used in the present study for generating the data presented in Fig. 5.

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Zhu, H., Bui, A., Santo, A. et al. 3H-1,2-dithiole-3-thione suppresses LPS-induced proinflammatory responses in macrophages: potential involvement of antioxidant induction, NF-κB, and Nrf2. Mol Cell Biochem 477, 1499–1506 (2022). https://doi.org/10.1007/s11010-021-04331-x

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