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Updated: Jul 24, 2025
Perturbations of Circulating miRNAs in Irritable Bowel Syndrome Detected Using a Multiplexed High-throughput Gene Expression Platform
Published on: November 30, 2016
MicroRNA-29b-3p promotes intestinal permeability in IBS-D via targeting TRAF3 to regulate the NF-κB-MLCK signaling
Yongfu Wang1, Wei Ke2,3,4, Jianfeng Gan5
1Department of Traditional Chinese Medicine, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China.
Abstract:
Irritable bowel syndrome with predominant diarrhea (IBS-D) is characterized by increased intestinal permeability. Previous studies have shown that the microRNA-29 gene is involved in the regulation of intestinal permeability in patients with IBS-D. NF-κB was proved to play a key role in inflammatory response of intestine and resultant disruption of tight junction integrity, whose activity could be inhibited by TNF Receptor-Associated Factor 3 (TRAF3). However, the exact mechanism that induces increased intestinal permeability in IBS-D patients has not been clarified. In this study, we found that microRNA-29b‑3p (miR-29b-3p) was significantly upregulated, while TRAF3 was decreased and the NF-κB-MLCK pathway was activated within the colonic tissue of IBS-D patients. Subsequently, we confirmed the targeting relationship between miR-29b-3p and TRAF3 through a double-luciferase reporter assay. Lentivirus transfection of NCM460 cells with miR-29b-3p-overexpressing and -silencing vectors demonstrated that the expression of TRAF3 was negatively correlated with the level of miR-29b-3p. The NF-κB/MLCK pathway was activated in the miR-29b-3p-overexpressing group and inhibited to some extent in the miR-29b-3p-silencing group. Results in WT and miR-29 knockout mice showed that miR-29b-3p levels were increased, TRAF3 levels were decreased, and the NF-κB/MLCK signaling was activated in the WT IBS-D group as compared with the WT control group. The protein levels of TRAF3 and TJs in the miR-29b-/- IBS-D group were partially recovered and NF-κB/MLCK pathway indicators were, to a certain extent, decreased as compared with the WT IBS-D group. These results suggested that miR-29b-3p deletion enhances the TRAF3 level in IBS-D mice and alleviates the high intestinal permeability. In brief, through the analysis of intestinal tissue samples from IBS-D patients and miR-29b-/- IBS-D mice, we showed that miR-29b-3p is involved in the pathogenesis of intestinal hyperpermeability in IBS-D via targeting TRAF3 to regulate the NF-κB-MLCK signaling pathway.
Insights
This study reveals that microRNA-29b-3p (miR-29b-3p) exacerbates intestinal hyperpermeability in Irritable Bowel Syndrome with Diarrhea (IBS-D) by downregulating TRAF3 and activating the NF-κB-MLCK pathway. Reducing miR-29b-3p levels may offer a therapeutic strategy for IBS-D.
Area of Science:
- Gastroenterology
- Molecular Biology
- Immunology
Background:
- Irritable Bowel Syndrome with predominant Diarrhea (IBS-D) is linked to increased intestinal permeability.
- The microRNA-29 gene family and the NF-κB pathway are implicated in IBS-D pathogenesis.
- TNF Receptor-Associated Factor 3 (TRAF3) can inhibit NF-κB activity, but its precise role in IBS-D-related hyperpermeability is unclear.
Purpose of the Study:
- To elucidate the mechanism underlying increased intestinal permeability in IBS-D.
- To investigate the role of microRNA-29b-3p (miR-29b-3p) and TRAF3 in IBS-D.
- To examine the involvement of the NF-κB-MLCK signaling pathway.
Main Methods:
- Analysis of colonic tissue from IBS-D patients and control subjects.
- In vitro studies using lentivirus transfection in NCM460 cells to manipulate miR-29b-3p levels.
- In vivo studies using wild-type and miR-29 knockout mice models of IBS-D.
- Double-luciferase reporter assay to confirm targeting relationship.
Main Results:
- miR-29b-3p was upregulated, TRAF3 downregulated, and the NF-κB-MLCK pathway activated in IBS-D patient colonic tissue.
- miR-29b-3p directly targets TRAF3, with inverse correlation observed between their expression levels.
- In miR-29 knockout mice with IBS-D, TRAF3 levels were restored, NF-κB-MLCK pathway activity reduced, and intestinal permeability alleviated compared to wild-type IBS-D mice.
Conclusions:
- miR-29b-3p contributes to intestinal hyperpermeability in IBS-D by targeting TRAF3 and activating the NF-κB-MLCK pathway.
- Modulation of miR-29b-3p levels offers a potential therapeutic avenue for IBS-D.
- The findings provide novel insights into the molecular mechanisms driving IBS-D pathophysiology.
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