Veratric acid relieves oxidative stress and DSS-induced colitis in mice by activating the Nrf2/HO-1 signaling pathway

doi:10.12122/j.issn.1673-4254.2026.02.18

Abstract

Abstract:

Objective To investigate the molecular mechanism by which veratric acid (VA) ameliorates oxidative stress injury and intestinal barrier dysfunction in mice with dextran sulfate sodium (DSS)-induced colitis. Methods Thirty male C57BL/6 mice were randomized equally into control group, DSS model group, and VA treatment group. The mice were assessed for changes in body weight, disease activity index (DAI), colon length, and colonic histopathology. Colonic expressions of TNF-α, IL-6, and IL-10 and oxidative stress markers (SOD, GSH, MDA, and COX-2) were determined using ELISA, and the expressions of tight junction proteins (ZO-1 and claudin-1) and Nrf2/HO-1 pathway proteins were detected using immunofluorescence staining and Western blotting. In Caco-2 cells with H₂O₂-induced oxidative stress, ROS accumulation was examined using flow cytometry and a DCFH-DA probe, and Nrf2 inhibitor (ML385) was used to validate the mechanism of VA for ameliorating oxidative stress. Results VA treatment significantly alleviated DSS-induced body weight loss, colon shortening and the increase of DAI score of the mice, resulting also in improved crypt structure and increased expressions of ZO-1 and claudin-1 and the number of goblet cells. VA obviously reduced colonic levels of TNF‑α and IL-6, increased the level of IL-10, and reversed DSS-induced decreases in SOD and GSH activity and increases in MDA and COX-2 levels. In H₂O₂-treated Caco-2 cells, VA decreased ROS-positive cell rate and intracellular ROS accumulation, and increased cellular expressions of claudin-1 and ZO-1. Mechanistically, VA promoted the expressions of Nrf2 and the downstream HO-1 protein, and ML385 partially reversed ROS-reducing effect of VA. Conclusion VA enhances antioxidant defense, inhibits inflammation, and repairs intestinal barrier function in mice with DSS-induced colitis by activating the Nrf2/HO-1 pathway, suggesting a novel strategy for treatment of inflammatory bowel disease.

Key words: inflammatory bowel disease, veratric acid, Nrf2/HO-1 signaling pathway, oxidative stress, intestinal barrier repair

Lin YIN, Keni ZHANG, Tong QIAO, Minzhu NIU, Lixia YIN, Xinyue LIU, Zhijun GENG, Jing LI, Jianguo HU. Veratric acid relieves oxidative stress and DSS-induced colitis in mice by activating the Nrf2/HO-1 signaling pathway[J]. Journal of Southern Medical University, 2026, 46(2): 403-411.

Figures/Tables 8

Fig.1 Effect of VA on DSS-induced colitis symptoms in mice (n=10). A: Changes of body weight. B: Changes of DAI scores. C, D: Comparison of colon length among different groups. *P<0.05 vs WT group; #P<0.05 vs DSS group.

Fig.2 Effect of VA on intestinal histopathology in DSS-treated mice (n=10). A: HE staining and inflammatory scores of the intestinal tissues of mice in each group. B: Histopathological scores of mice in each group. C: AB-PAS staining of the intestinal tissues in each group. D: Comparison of goblet cell counts in each group. *P<0.05 vs WT group; #P<0.05 vs DSS group.

Fig.3 Effect of VA on expressions of inflammatory cytokines in the colonic tissues of DSS-treated mice (n=10). A: Levels of interleukin-6 (IL-6) in the colonic mucosa of the mice in each group detected by enzyme-linked immunosorbent assay (ELISA). B: Levels of interleukin-10 (IL-10) in the colonic mucosa of the mice in each group detected by ELISA. C: Levels of tumor necrosis factor-α (TNF-α) in the colonic mucosa of the mice in each group detected by ELISA. *P<0.05 vs WT group; #P<0.05 vs DSS group.

Fig.4 Effect of VA on tight junction proteins in the colonic tissues of DSS-induced mice (n=10). A: Immunofluorescence staining for claudin-1 and ZO-1 in the colons of the mice in each group. B: Western blotting for detecting expression levels of claudin-1 and ZO-1 in the colons of the mice in each group. *P<0.05 vs WT group; #P<0.05 vs DSS group.

Fig.5 Regulatory effect of VA on oxidative stress in the colonic tissues of DSS-induced mice (n=10). A: Detection results of glutathione (GSH) in each group. B: levels of superoxide dismutase (SOD) in each group. C: Malondialdehyde (MDA) levels in each group. D: Cyclooxygenase-2 (COX-2) levels in each group. *P<0.05 vs WT group; #P<0.05 vs DSS group.

Fig.6 Effect of VA on reactive oxygen species (ROS) production induced by H₂O₂ in Caco-2 cells (n=10). A: ROS staining with the fluorescent probe (DCFH-DA) in each group. B: ROS detection by flow cytometry in each group. *P<0.05 vs Control group. #P<0.05 vs H₂O₂ group.

Fig.7 Effect of VA on intestinal barrier function of H₂O₂-induced Caco-2 cells (n=10). A: Immunofluorescence staining for claudin-1 and ZO-1 in each group. B: Western blotting for detecting expression levels of claudin-1 and ZO-1 in each group. *P<0.05 vs Control group; #P<0.05 vs H₂O₂ group.

Fig.8 VA alleviates intestinal oxidativestress in colitis mice by activating the Nrf2/HO-1 pathway. A: Immunofluorescencestaining of Nrf2/HO-1 in the colons of themice in each group. B: Intracellular ROSdetection using the fluorescent probe(DCFH-DA) in VA treatment group andML385 inhibitor group. C: IntracellularROS detection by flow cytometry in VAtreatment group and ML385 inhibitorgroup. D: Western blotting for detectingcolonic expression levels of Nrf2/HO-1 ofthe mice in each group. E: Western blottingfor detecting esxpression levels of Nrf2/HO-1 in Caco-2 cells in each group. *P<0.05vs (WT/VA-C/control) group. #PP<0.05 vs(DSS/H₂O₂) group.

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