Disruption of ZO-1/claudin-4 interaction in relation to inflammatory responses in methotrexate-induced intestinal mucositis View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2013-08-21

AUTHORS

Kazuma Hamada, Naoko Kakigawa, Shuichi Sekine, Yoshihisa Shitara, Toshiharu Horie

ABSTRACT

PurposeMethotrexate (MTX)-induced intestinal mucositis limits the use of the drug. We previously reported that MTX-dependent production of reactive oxygen species is an initiating signal leading to neutrophil migration and intestinal barrier dysfunction. Moreover, alterations of zonula occludens (ZO)-1, an integral component of tight junctions (TJs), contribute to its dysfunction. This study aimed to clarify the identity of inflammatory mediators in the intestine of MTX-treated rats and to evaluate MTX-stimulated alterations in the expression of TJ proteins other than ZO-1 (e.g., occludin and claudins).MethodsMale Wistar rats were administrated MTX (15 mg kg−1) orally once daily for 4 days. Tumor necrosis factor (TNF)-α, interleukin (IL)-1β, macrophage inflammatory protein (MIP)-2, cytokine-induced neutrophil chemoattractant-2, Toll-like receptor 4 (TLR4), and occludin were determined by real-time RT-PCR. Expression, distribution, and interactions of TJ proteins were evaluated by Western blotting, immunohistochemistry, and immunoprecipitation.ResultsMTX increased the mRNA levels of TNF-α, IL-1β, MIP-2, and TLR4 in the small intestine, as well as the protein expression of claudin-2. Increased claudin-2 and decreased claudin-4 immunostaining were also observed. Occludin mRNA levels were significantly diminished by MTX administration, whereas occludin protein levels and the interaction between ZO-1 and occludin were unaltered; however, the interaction between ZO-1 and claudin-4 was significantly compromised.ConclusionsThese results indicate that elevated levels of inflammatory cytokines and chemokines in the small intestine of MTX-treated rats may contribute to the inhibition of ZO-1/claudin-4 binding, and that inhibition of ZO-1/claudin-4 binding may in turn lead to a reduction in claudin-4 expression. More... »

PAGES

757-765

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/s00280-013-2238-2

DOI

http://dx.doi.org/10.1007/s00280-013-2238-2

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1048016249

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/23963446


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