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Glycyrrhizin ameliorates experimental colitis through attenuating interleukin-17-producing T cell responses via regulating antigen-presenting cells

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Abstract

Glycyrrhizin, a component of Chinese medicine licorice root, has the ability to inhibit the functions of high-mobility group box 1 (HMGB1). While glycyrrhizin is known to have anti-inflammatory activities, the underlying mechanisms by which glycyrrhizin inhibits inflammation during the development of trinitrobenzenesulfonic acid (TNBS)-induced experimental colitis are not well understood. This study systemically examined the regulatory effects of glycyrrhizin on inflammatory response in TNBS-induced murine colitis and explored the potential mechanisms involved in this process. We reported that glycyrrhizin treatment ameliorated colitis and decreased the production of inflammatory mediators HMGB1, IFN-γ, IL-6, TNF-α, and IL-17. In addition, glycyrrhizin regulated responses of dendritic cells (DCs) and macrophages during the development of colitis. Furthermore, administration of glycyrrhizin suppressed the proliferation of Th17 cells in colitis. Moreover, the ability of DCs and macrophages to induce the differentiation of Th17 cells was enhanced in presence of HMGB1, which was inhibited by glycyrrhizin. These results demonstrated that glycyrrhizin alleviated colitis by inhibiting the promotive effect of HMGB1 on DC/macrophage-mediated Th17 proliferation. In conclusion, HMGB1 plays an important role in the development of colitis. As an inhibitor of HMGB1, glycyrrhizin might be a novel therapy for colitis.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (91542110 to M. Fang), the Ministry of Science and Technology of China (Grant No. 2013CB530505), and the National Natural Science Foundation of China (81373167 to M. Fang).

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Authors and Affiliations

  1. Department of Immunology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, 13# Hangkong Road, Wuhan, 430030, People’s Republic of China

    Xiangyu Chen, Lingyun Li, Liyong Chen, Qirui Li, Feili Gong & Min Fang

  2. Faculty of Medicine, Hubei University of Science and Technology, Xianning, People’s Republic of China

    Dai Fang

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  1. Xiangyu Chen
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Correspondence to Min Fang.

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Electronic supplementary material

Figure s1

Glycyrrhizin regulated the percentages of IFN-γ+CD4+ T help (Th1) cells and IL-4+CD4+ T help (Th2) cells in the development of colitis. Primary lymphocytes were isolated from the spleen, mesenteric lymph nodes (MLN), colonic lamina propria (cLP) of mice. FACS analysis was performed after staining of anti-CD3, CD4, IFN-γ and IL-4 antibodies. (a) Gating strategies for Th17 cells were illustrated. (b) The percentages of IFN-γ+/CD3+CD4+ T cells in spleen, MLN and cLP were determined. (c) The percentages of IL-4+/CD3+CD4+ T cells in the spleen, MLN and cLP were determined. The cells were gated on CD3+CD4+ T cells. Data represent the mean ± SD, *p≤0.05. The experiments were replicated at least twice. (GIF 42 kb)

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Figure s2

Glycyrrhizin regulated the percentages of FoxP3+CD4+ regulatory T (Treg) cells in the development of colitis. Primary lymphocytes were isolated from the spleen, mesenteric lymph nodes (MLN), colonic lamina propria (cLP) of mice. FACS analysis was performed after staining of anti-CD4 and FoxP3 antibodies. (a) Gating strategies for Treg cells were illustrated. (b) The percentages of FoxP3+CD4+ T cells in the spleen, MLN and cLP were determined. Data represent the mean ± SD, *p≤0.05. The experiments were replicated at least twice. (GIF 23 kb)

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Figure s3

Glycyrrhizin inhibited the ability of BMDCs/BMDMs to induce the differentiation of Th1/Th2 cells in the presence or absence of HMGB1. Naive CD4+ T cells were sorted by immunomagnetic selection from spleens. Naive CD4+ T cells (1 × 105) were cultured alone or co-cultured with (a) BMDCs / (b) BMDMs (2.5 × 104) which were treated with or without 100 ng/mL HMGB1 in the presence or absence of glycyrrhizin/anti-HMGB1 antibody for 4 days, and the IFN-γ+CD4+ T cells and IL-4+CD4+ T cells were analyzed by using flow cytometry. Data represent the mean ± SD, *p≤0.05. The experiments were replicated at least twice. (GIF 43 kb)

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Figure s4

Flow cytometry scatter plot. (a) Correspond to figure 3b; (b) Correspond to figure 3c. (GIF 85 kb)

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Figure s5

Flow cytometry scatter plot. (a) Correspond to figure 4b; (b) Correspond to figure 4c and d. (GIF 61 kb)

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Figure s6

Flow cytometry scatter plot correspond to figure 5b. (GIF 24 kb)

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Figure s7

Flow cytometry scatter plot. (a) Correspond to figure 6a; (b) Correspond to figure 6b. (GIF 105 kb)

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Figure s8

Flow cytometry scatter plot. (a) Correspond to figure 7a; (b) Correspond to figure 7b. (GIF 50 kb)

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Figure s9

Flow cytometry scatter plot. (a) Correspond to figure s1b; (b) Correspond to figure s1c. (GIF 47 kb)

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Figure s10

Flow cytometry scatter plot correspond to figure s2b. (GIF 48 kb)

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Figure s11

Flow cytometry scatter plot. (a) Correspond to figure s3a; (b) Correspond to figure s3b. (GIF 89 kb)

High resolution image (TIFF 1316 kb)

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Chen, X., Fang, D., Li, L. et al. Glycyrrhizin ameliorates experimental colitis through attenuating interleukin-17-producing T cell responses via regulating antigen-presenting cells. Immunol Res 65, 666–680 (2017). https://doi.org/10.1007/s12026-017-8894-2

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