GASTROENTEROLOGY / CLINICAL RESEARCH
STAT1/MUC4 activation promotes antimicrobial peptide production to reduce intestinal epithelium barrier injury caused by enteropathogenic Escherichia coli infection
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1
Department of Laboratory, The First Hospital of Qinhuangdao, Hebei, China
2
Department of Critical Care Medicine, The First Hospital of Qinhuangdao, Hebei, China
Submission date: 2023-05-04
Final revision date: 2023-08-31
Acceptance date: 2023-09-01
Online publication date: 2023-09-03
Corresponding author
Zhigang Zuo
Department of Critical
Care Medicine
The First Hospital
of Qinhuangdao
No. 258 Wenhua Road
Haigang District
Qinhuangdao City
Hebei Province, China
KEYWORDS
TOPICS
ABSTRACT
Introduction:
Antimicrobial peptides (AMPs) are endogenous peptides that have been identified to alleviate intestinal epithelial barrier inflammation and dysfunction caused by enteropathogenic Escherichia coli (EPEC) infection; nonetheless, the upstream molecular mechanism of the production of AMPs is poorly understood.
Material and methods:
The binding of signal transducer and activator of transcription (STAT) 1 (STAT1) to mucin 4 (MUC4) was examined by co-immunoprecipitation assay. To detect the influence of STAT1 and MUC4 expression, a C57BL/6 mouse model of EPEC infection in vivo and an EPEC infected intestinal epithelial cell (IEC) in vitro model were established. Expression levels of STAT1, MUC4, phosphorylated (p)-STAT1, proinflammatory cytokines, zonula occludens-1 (ZO-1) and AMP-related genes in mouse ileum and/or IEC were analyzed by immunohistochemical test, immunofluorescence assay, Western blot, and/or qRT-PCR. Meanwhile, IEC viability and apoptosis were measured using CCK-8 assay and flow cytometry.
Results:
p-STAT1, MUC4, ZO-1 and AMP-related genes were lowly expressed in the ileum of EPEC-infected mice. p-STAT1 and MUC4 bound to each other. The expression levels of STAT1 and MUC4 were decreased in EPEC-infected IEC. STAT1 overexpression counteracted the EPEC-induced reduction of viability, apoptosis promotion, ZO-1 activity inhibition, release of proinflammatory cytokines, and downregulation of MUC4 and AMP-related genes in IEC. MUC4 knockdown partly counteracted the effect of STAT1 overexpression, but did not affect the forced STAT1 overexpression in EPEC-infected IEC.
Conclusions:
STAT1/MUC4 pathway activation promotes AMP production to mitigate intestinal epithelium barrier injury caused by EPEC infection.
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