Niranthin ameliorates Crohn's disease-like colitis in mice via antagonizing intestinal epithelial cell apoptosis and regulating intestinal Th1/Th2 immune homeostasis

doi:10.12122/j.issn.1673-4254.2026.03.19

Abstract

Abstract:

Objective To investigate the effect of niranthin (Nir) in mice with Crohn's disease (CD)‑like colitis and its therapeutic mechanism. Methods In a mouse model of CD-like colitis induced using 2,4,6-trinitrobenzene sulfonic acid, the effects of niranthin on colitis symptoms were evaluated by measuring changes in disease activity index (DAI) score, body weight, colon length, and colonic pathologies. Intestinal barrier function and cell apoptosis were evaluated using AB-PAS staining, immunofluorescence staining, Western blotting, and TUNEL staining. The changes in Th1 and Th2 cells in the mesenteric lymph nodes and colonic TNF-α and IL-10 expression levels were determined with flow cytometry and ELISA. In lipopolysaccharide (LPS)-induced mouse colon organoids, the effects of niranthin on organoid budding number and barrier protein expressions were observed. Network pharmacology and in vivo experiments were employed to explore and verify the therapeutic mechanism of niranthin on colitis. Results In the mouse models of CD-like colitis, niranthin treatment obviously improved weight loss, DAI scores, and colorectal shortening and significantly reduced tissue inflammation scores, goblet cell loss, and intestinal epithelial cell apoptosis. Niranthin significantly increased the budding number in LPS-induced mouse colon organoids. In both mouse colon tissues and LPS-induced mouse colon organoids, niranthin obviously increased the expressions of ZO-1 and claudin-1, downregulated the expressions of cleaved caspase-3 and BAX, and upregulated Bcl‑2 expression. The niranthin-treated mouse models showed significantly ameliorated Th1/Th2 imbalance in the mesenteric lymph nodes, downregulated TNF‑α and upregulated IL‑10 levels in the colon tissues. Network pharmacology predicted that the therapeutic mechanism of niranthin for CD-like colitis involved the PI3K/AKT pathway, which was validated in the mouse models treated with niranthin and the PI3K/AKT pathway activator Recilisib. Conclusion Niranthin ameliorates colitis in mice by antagonizing epithelial apoptosis and regulating Th1/Th2 balance via inhibiting the PI3K/AKT pathway.

Key words: Crohn's disease, niranthin, intestinal barrier, apoptosis, Th1/Th2, PI3K/AKT

Hexin WEN, Jie LIN, Lugen ZUO, Mulin LIU. Niranthin ameliorates Crohn's disease-like colitis in mice via antagonizing intestinal epithelial cell apoptosis and regulating intestinal Th1/Th2 immune homeostasis[J]. Journal of Southern Medical University, 2026, 46(3): 655-665.

Figures/Tables 8

Fig.1 Effects of niranthin (Nir) intervention in mice with TNBS-induced colitis. A: Body weight changes. B: DAI scores. C, D: Colon length. E, G: Inflammation scores and HE staining of mouse colon tissues. F, H: Number of goblet cells and AB-PAS staining. n=6, *P<0.05 vs WT; #P<0.05 vs TNBS.

Fig.2 Effects of Nir intervention on intestinal barrier function in TNBS-induced colitis mice. A: Immunofluorescence assay of ZO-1 and claudin-1 expression. B, C: Western blotting for claudin-1 and ZO-1 expressions. n=6, *P<0.05 vs WT; #P<0.05 vs TNBS.

Fig.3 The effect of Nir intervention on colonic cell apoptosis in TNBS-induced colitis mice. A: TUNEL staining. B, C: Western blotting for C-caspase-3, Bax and Bcl-2 expressions. n=6, *P<0.05 vs WT; #P<0.05 vs TNBS.

Fig.4 Effect of Nir on LPS-induced intestinal epithelial cell apoptosis in mouse colon organoids. A: Representative images of colon organoids. B: Number of organoids budding. C, D: Western blotting and bar chart for C-caspase-3, Bax and Bcl-2 expressions. n=6, *P<0.05 vs Con; #P<0.05 vs LPS.

Fig.5 Effects of Nir intervention on intestinal barrier function in LPS-induced colonic organoids. A, B: Immunofluorescence assay of ZO-1 and claudin-1 expression. C, D: Western blotting for claudin-1 and ZO-1 expressions. n=6, *P<0.05 vs Con; #P<0.05 vs LPS.

Fig.6 Effects of Nir intervention on Th1/Th2 balance in mesenteric lymph nodes and inflammatory factor levels in colonic tissues of TNBS-induced colitis mice. A, B: Proportion of Th1 cells in the mesenteric lymph nodes. C, D: Proportion of Th2 cells in the mesenteric lymph nodes. E: Th1/Th2 ratio. F, G: Expression levels of intestinal mucosal inflammatory mediators. n=6, *P<0.05 vs WT; #P<0.05 vs TNBS.

Fig.7 Ameliorative effect of Nir on colitis is possibly associated with the PI3K/AKT pathway. A: Venn diagram. B: KEGG enrichment analysis. C: GO enrichment analysis. D, F: Western blotting of p-PI3K, PI3K, p-AKT and AKT expressions in the colon tissue. E, G: Western blotting of p-PI3K, PI3K, p-AKT and AKT expressions in the colonic organoids. n=6, *P<0.05 vs WT; #P<0.05 vs TNBS.

Fig.8 Nir can restore immune homeostasis and barrier integrity by negatively regulating the PI3K/AKT pathway. A, C: Western blotting of p-PI3K, PI3K, p-AKT and AKT expressions. B, D: Western blotting for C-caspase-3, Bax and Bcl-2 expressions. E, F: Proportion of Th1 cells in the mesenteric lymph nodes. G, H: Proportion of Th2 cells in the mesenteric lymph nodes. I: Th1/Th2 ratio. J, K: Western blotting for p-p65 and p65 expressions. n=6, *P<0.05 vs TNBS; #P<0.05 vs Nir.

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