ReferenceID 3168

Wogonoside alleviates colitis by improving intestinal epithelial barrier function via the MLCK/pMLC2 pathway

Phytomedicine

BACKGROUND: Intestinal epithelial barrier dysfunction, which involves myosin light chain kinase (MLCK) activation, contributes to the occurrence and progression of inflammation in inflammatory bowel disease (IBD). Wogono

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Reference Id
3168
Evidence Id
19758
Core Evidence Id
19758
Source Reference Id
6317
Herb2 Reference Id
HBREF007114
Subject Paper Key
HBIN048376_32062328
Pubmed Id
32062328
Doi
10.1016/j.phymed.2020.153179
Paper Title
Wogonoside alleviates colitis by improving intestinal epithelial barrier function via the MLCK/pMLC2 pathway
Paper Abstract
BACKGROUND: Intestinal epithelial barrier dysfunction, which involves myosin light chain kinase (MLCK) activation, contributes to the occurrence and progression of inflammation in inflammatory bowel disease (IBD). Wogonoside helps maintain intestinal homeostasis in mice with dextran sulfate sodium (DSS)-induced colitis, but it is unclear whether it modulates intestinal barrier function. PURPOSE: Here, we demonstrate that wogonoside protects against intestinal barrier dysfunction in colitis via the MLCK/pMLC2 pathway both in vivo and in vitro. METHODS: Caco-2 cell monolayers treated with the proinflammatory cytokine TNF-alpha showed barrier dysfunction and were assessed in the absence and presence of wogonoside for various physiological, morphological, and biochemical parameters. Colitis was induced by 3% DSS in mice, which were used as an animal model to explore the pharmacodynamics of wogonoside. We detected MLCK/pMLC2 pathway proteins via western blot analysis, assessed the cytokines IL-13 and IFN-gamma via ELISA, tested bacterial translocation via fluorescence in situ hybridization (FISH) and a proper sampling of secondary lymphoid organs for bacterial culture. In addition, the docking affinity of wogonoside and MLCK was observed with DS2.5 software. RESULTS: Wogonoside alleviated the disruption of transepithelial electrical resistance (TER) in TNF-alpha exposured Caco-2 cell; FITC-dextran hyperpermeability; loss of the tight junction (TJ) proteins occludin, ZO-1 and claudin-1 in Caco-2 cell monolayers; and bacterial translocation in colitic mice. Moreover, wogonoside reduced the levels of the proinflammatory cytokines IL-13 and IFN-gamma to maintain intestinal immune homeostasis. Transmission electron microscopy (TEM) confirmed that wogonoside ameliorated the destruction of intestinal epithelial TJs. Wogonoside not only inhibited the cytoskeletal F-actin rearrangement induced by TNF-alpha, stabilized the cytoskeletal structure, suppressed MLCK protein expression, and reduced MLC2 phosphorylation. In addition, the results of molecular docking analysis showed that wogonoside had a high affinity for MLCK and formed hydrogen bonds with the amino acid residue LYS261 and pi bonds with LYS229. CONCLUSION: Collectively, our study indicates that wogonoside alleviates colitis by protecting against intestinal barrier dysfunction, and the potential mechanism may involve regulation of TJs via the MLCK/pMLC2 signaling pathway. Meanwhile, our study also explains the success of S. baicalensis in the treatment of ulcerative colitis (UC).
Journal
Phytomedicine
Publish Year
2020
Experiment Subject
mouse; caco-2 cell monolayers; tnf-alpha exposured caco-2 cell
Experiment Type
Animal & Cell Experiment
Phenotype Related
Ulcerative Colitis; Inflammation; Inflammatory Bowel Disease; Intestinal Epithelial Barrier Dysfunction; Colitis; Intestinal Barrier Dysfunction
Paper Title Cn
Paper Title En
Wogonoside alleviates colitis by improving intestinal epithelial barrier function via the MLCK/pMLC2 pathway
Bilingual Status
semi_complete