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Carbon Monoxide-Releasing Molecule-3 Regulates the Polarization of Lipopolysaccharide-Induced Macrophages

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Abstract

Macrophages show two main phenotypes, the M1-type (pro-inflammatory) and the M2-type (anti-inflammatory). The purpose of this research was to investigate the regulatory effect of carbon monoxide releasing molecule-3 (CORM-3) on LPS-induced macrophage polarization. LPS-induced RAW264.7 cells were exposed to CORM-3 for 24 h. Polarization of cells was checked by flow cytometry; expression of M1 or M2 macrophage-related factors and NF-κB signaling factors was examined by RT-PCR, ELISA, and Western blot. Male C57 mice were divided into three groups: normal group; periodontitis group, where experimental periodontitis was established in mice; LPS+CORM-3 group, where mice with experimental periodontitis were treated with CORM-3. Polarization of macrophages and the expression of M1 or M2 macrophage-related factors were detected by immunofluorescence, ELISA, and RT-PCR. CORM-3 significantly reduced M1 macrophage proportion, but increased M2 proportion in LPS-stimulated cells. Accordingly, CORM-3 significantly suppressed the expression of M1 macrophage-related TNF-α, iNOS, IL-1β, and IL-6, but promoted M2-related IL-10 and Arg-1. The expression of p-p65, p-p50, and p-IκB induced with LPS was inhibited by CORM-3. In vivo experiments indicated that CORM-3 induced more M2 macrophages in periodontal tissues in mice with experimental periodontitis. The expression of M1 macrophage-related factor in periodontitis was inhibited, but the expression of M2-related factors was increased by CORM-3. CORM-3 inhibits macrophage polarization to pro-inflammatory M1-type and promotes to anti-inflammatory M2-type, which provides scientific basis for the application of CORM-3 in the treatment of periodontitis.

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Availability of Data and Material

The data used to support the findings of this study are available from the corresponding author upon request.

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Funding

This work was supported by Shandong Provincial Natural Science Foundation (grant number [ZR2020MH186]), Jinan Science and Technology Innovation Program in Clinical Medicine (grant numbers [201805045]), and Shandong Provincial Science and Technology Development Plan (grant number [2010GSF10270]).

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Authors and Affiliations

  1. Department of Vip Center, School and Hospital of Stomatology & Shandong Provincial Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, Cheeloo College of Medicine, Shandong University, No.44-1 Wenhua Road West, Jinan, 250012, Shandong, China

    Tingting Liu, Qingbin Han, Jingyuan Li & Hui Song

  2. Department of Stomatology, Linyi People’s Hospital, Linyi, Shandong, China

    Qingbin Han

  3. Yantai Stomatological Hospital, Yantai, Shandong, China

    Yan Pan

Authors
  1. Tingting Liu
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  2. Qingbin Han
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  3. Yan Pan
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  4. Jingyuan Li
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  5. Hui Song
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Contributions

All authors contributed to the study conception and design. Material preparation, data collection, and analysis were performed by Tingting Liu; Qingbin Han and Yan Pan participated in the in vivo procedures. The first draft of the manuscript was written by Tingting Liu; Jingyuan Li and Hui Song have been involved in analyzing the data and revising the manuscript critically. All authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Hui Song.

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Approval was obtained from the ethics committee of Shandong University School of Stomatology. The procedures used in this study adhere to the tenets of the Declaration of Helsinki.

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Liu, T., Han, Q., Pan, Y. et al. Carbon Monoxide-Releasing Molecule-3 Regulates the Polarization of Lipopolysaccharide-Induced Macrophages. Inflammation 44, 1737–1749 (2021). https://doi.org/10.1007/s10753-021-01450-x

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  • DOI: https://doi.org/10.1007/s10753-021-01450-x

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