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May 20, 2026Journal of Biochemical and Molecular Toxicology1 citations

The PVT1/miR‐216a‐5p/TLR4 Signaling Axis Regulates the Inflammation and Intestinal Epithelial Barrier Function of Ulcerative Colitis

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MLMeixiao LiuZXZhiguang XuLZLinqi Zhang

Key Points

  • This research aims to understand how the PVT1/miR-216a-5p/TLR4 signaling axis influences inflammation and intestinal barrier integrity in ulcerative colitis.
  • LPS-stimulated RAW264.7 cells co-cultured with Caco-2 cells were used to model UC in vitro.
  • C57BL/6 J mice induced by 3% DSS served as the in vivo model.
  • Cytotoxicity, barrier function, pro-inflammatory factors, and gene expression levels were evaluated using various assays.
  • Increased PVT1 and TLR4 levels were observed while miR-216a-5p levels decreased in LPS-treated cells and DSS mice.
  • Higher levels of pro-inflammatory factors were detected in RAW264.7 and DSS mouse colon tissue, with decreased ZO-1 and occludin levels in Caco-2 cells.
  • Inhibiting PVT1 expression reversed the inflammatory effects, while miR-216a-5p inhibition countered the protective effects of PVT1 knockdown.

Abstract

Ulcerative colitis (UC) is a chronic, relapsing disease, imposing a heavy disease burden and severely reducing the quality life of patients. This study verified the influence of the PVT1/miR-216a-5p/TLR4 signaling axis on UC. LPS-stimulated RAW264.7 cells were co-cultured with Caco-2 cells to establish an in vitro model. The C57BL/6 J mice induced by 3% DSS were used as an in vivo model. Cytotoxicity and barrier function were evaluated in Caco-2 cells. The PVT1, miR-216a-5p, and TLR4 levels were detected by RT-qPCR. ELISA was used to detect the level of pro-inflammatory factors in RAW264.7 supernatant and colon tissue, and the level of ZO-1 and occludin in Caco-2 cells and colon tissue. TEER was used to evaluate the integrity of the barrier function of Caco-2 cells. The targeting relationships between PVT1, miR-216a-5p, and TLR4 were verified by dual-luciferase reporter gene assay. The PVT1 and TLR4 levels were increased, but miR-216a-5p was decreased in the LPS-treated RAW264.7, co-cultured Caco-2, and colon tissue of DSS mice. The pro-inflammatory factor levels in RAW264.7 and colon tissue of DSS mice were increased, while in Caco-2, the cytotoxicity was aggravated, and the integrity of the barrier function was disrupted. The levels of ZO-1 and occludin were decreased in Caco-2 cells and tissue. If the PVT1 expression was inhibited, the results showed an opposite trend. In contrast, inhibition of miR‑216a‑5p reversed the protective effects induced by PVT1 knockdown. The PVT1/miR-216a-5p signaling axis affects the inflammation and intestinal epithelial cell barrier function in UC, which may be a possible therapeutic target for UC.

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Cite This Study

Liu et al. (2026) studied this question.

synapsesocial.com/papers/6a0d5089f03e14405aa9c5d3https://doi.org/10.1002/jbt.70887
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