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INFECTIOUS DISEASES / BASIC RESEARCH
STAT1/MUC4 activation promotes antimicrobial peptide production to reduce intestinal epithelium barrier injury caused by enteropathogenic Escherichia coli infection
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
Department of Critical Care Medicine The First Hospital of Qinhuangdao No. 258 Wenhua Road Haigang District Qinhuangdao City Hebei Province, China
Arch Med Sci 2025;21(3):1036-1050
KEYWORDS
antimicrobial peptideenteropathogenic Escherichia coli infectionintestinal epithelium barrier injurySTAT1/MUC4 pathwayZO-1
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.
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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| eISSN: | 1896-9151 |
| ISSN: | 1734-1922 |
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