2,6-dimethoxy-1,4-benzoquinone alleviates dextran sulfate sodium-induced ulcerative colitis in mice by suppressing NLRP3 inflammasome activation

doi:10.12122/j.issn.1673-4254.2025.08.10

Abstract

Abstract:

Objective To investigate the therapeutic mechanism of 2,6-dimethoxy-1,4-benzoquinone (DMQ) for alleviating dextran sulfate sodium (DSS)-induced ulcerative colitis (UC) in mice. Methods Eighteen male C57BL/6J mice were equally randomized into control group, DSS group and DMQ treatment group. In DSS and DMQ groups, the mice were treated with DSS in drinking water to induce UC, and received intraperitoneal injections of sterile PBS or DMQ (20 mg/kg) during modeling. The changes in body weight, disease activity index (DAI), colon length, spleen weight, and colon histological scores of the mice were examined, and the percentages of Th17 and IFN-γ⁺ CD8⁺ T cells in the mesenteric lymph nodes and spleen were analyzed using flow cytometry. The expressions of tight junction proteins (Occludin and ZO-1), proteins associated with inflammasome activation (caspase-1 and p20), IL-1β and TNF-α in the colon tissues were detected using Western blotting or ELISA. In the cell experiment, mouse bone marrow-derived macrophages (BMDMs) primed with lipopolysaccharide (LPS) were treated with DMQ, followed by stmulation with nigericin to activate the classical NLRP3 inflammasome pathway. In cultured human peripheral blood mononuclear cells (PBMCs) treated with either LPS alone or LPS plus nigericin, the effects of DMQ on inflammasome activation, pyroptosis, and cytokine release were evaluated via Western blotting, ELISA, and flow cytometry. Results In DSS-treated mice, DMQ treatment significantly alleviated DSS-induced body weight loss, colon shortening, spleen enlargement, and colon inflammation. The DMQ-treated mice showed significantly reduced percentages of Th17 cells and IFN-γ⁺ CD8⁺ T cells in the mesenteric lymph nodes and spleen, with increased occludin and ZO-1 expressions and decreased caspase-1 expression in the colon tissue. DMQ obviously inhibited classical NLRP3 inflammasome activation in mouse BMDMs and both the classical and alternative pathways of NLRP3 activation in human PBMCs, causing also suppression of caspase-1-dependent pyroptosis. Conclusion DMQ ameliorates DSS-induced UC in mice by inhibiting NLRP3 inflammasome activation.

Key words: 2,6-dimethoxy-1,4-benzoquinone, NLRP3 inflammasome, ulcerative colitis, natural products

Chenfei LIU, Wei ZHANG, Yao ZENG, Yan LIANG, Mengting WANG, Mingfang ZHANG, Xinyuan LI, Fengchao WANG, Yanqing YANG. 2,6-dimethoxy-1,4-benzoquinone alleviates dextran sulfate sodium-induced ulcerative colitis in mice by suppressing NLRP3 inflammasome activation[J]. Journal of Southern Medical University, 2025, 45(8): 1654-1662.

Figures/Tables 6

Fig.1 Effect of 2,6-dimethoxy-1,4-benzoquinone (DMQ) on body weight loss (A) and Disease Activity Index (DAI) score (B) in mouse models with dextran sulfate sodium (DSS)-induced ulcerative colitis (UC). **P<0.01, ***P<0.001.

Fig. 2 Effect of DMQ treatment on colon inflammation in mice with DSS-induced UC. A, B: Comparison of colon length among the groups. C: Comparison of spleen weight among the groups. D: HE staining of the colon tissue in each group. E: Histological scores of the colon tissue in each group. n=6. *P<0.05, ***P<0.001.

Fig.3 Effect of DMQ treatment on immune regulation in mice with DSS-induced UC. A: Flow cytometry for detection of the ratio of CD4+IL-17+ cells in the mesenteric lymph nodes of the mice. B: Ratios of CD4+IL-17+ cells in the spleen of the mice. C: Ratios of CD8+ IFN-γ+ cells in the mesenteric lymph nodes of the mice. D: Ratios of CD8+ IFN-γ+ cells in the spleen of the mice. *P<0.05, **P<0.01.

Fig.4 Effect of DMQ treatment on intestinal barrier integrity in mice with DSS-induced UC. A-C: Western blotting for detecting occludin and ZO-1 expressions in the colonic tissues. **P<0.01, ***P<0.001.

Fig.5 Effect of DMQ treatment on colonic NLRP3 inflammasome activation in mice with DSS-induced UC. A: Western blotting of p20 expression in the colonic tissue. B: ELISA for IL-1β in the colonic tissue. C: ELISA for TNF-α in the colonic tissue. **P<0.01, ***P<0.001.

Fig.6 Effect of DMQ treatment on NLRP3 inflammasome activation in the cell models. A: ELISA detection of IL-1β in the culture supernatant of nigericin-stimulated BMDMs. B,C: ELISA detection of IL-1β and TNF-α in the culture supernatant of nigericin-stimulated PBMCs. D,E: ELISA detection of IL-1β and TNF-α in the culture supernatant of LPS-stimulated PBMCs. **P<0.001, ***P<0.001.

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