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Original article
Nrf2 deficiency impairs the barrier function of mouse oesophageal epithelium
  1. Hao Chen1,
  2. Yuhui Hu1,
  3. Yu Fang1,2,
  4. Zorka Djukic3,
  5. Masayuki Yamamoto4,
  6. Nicholas J Shaheen3,
  7. Roy C Orlando3,
  8. Xiaoxin Chen1,3
  1. 1Cancer Research Program, Julius L Chambers-Biomedical Biotechnology Research Institute, North Carolina Central University, Durham, North Carolina, USA
  2. 2Department of Cardiovascular and Thoracic Surgery, The Second Xiangya Hospital, Central South University, Changsha, China
  3. 3Division of Gastroenterology and Hepatology, Department of Medicine, Center for Esophageal Diseases and Swallowing, University of North Carolina, Chapel Hill, North Carolina, USA
  4. 4Department of Medical Biochemistry, Tohoku University Graduate School of Medicine, Sendai, Japan
  1. Correspondence to Dr Xiaoxin Luke Chen, Cancer Research Program, Julius L Chambers Biomedical Biotechnology Research Institute, North Carolina Central University, 700 George Street, Durham, NC 27707, USA; lchen{at}nccu.edu

Abstract

Objective As a major cellular defence mechanism, the Nrf2/Keap1 pathway regulates expression of genes involved in detoxification and stress response. Here we hypothesise that Nrf2 is involved in oesophageal barrier function and plays a protective role against gastro-oesophageal reflux disease (GERD).

Design Human oesophageal biopsy samples, mouse surgical models and Nrf2−/− mice were used to assess the role of the Nrf2/Keap1 pathway in oesophageal barrier function. Trans-epithelial electrical resistance (TEER) was measured with mini-Ussing chambers. HE staining and transmission electron microscopy were used to examine tissue morphology, while gene microarray, immunohistochemistry, western blotting and chromatin immunoprecipitation (ChIP) analysis were used to assess gene expression.

Results Nrf2 was expressed in normal oesophageal epithelium and activated in GERD of both humans and mice. Nrf2 deficiency and gastro-oesophageal reflux in mice, alone or in combination, reduced TEER and increased intercellular space in oesophageal epithelium. Nrf2 target genes and gene sets associated with oxidoreductase activity, mitochondrial biogenesis and energy production were downregulated in the oesophageal epithelium of Nrf2−/− mice. Consistent with the antioxidative function of Nrf2, a DNA oxidative damage marker (8OHdG) dramatically increased in oesophageal epithelial cells of Nrf2−/− mice compared with those of wild-type mice. Interestingly, ATP biogenesis, Cox IV (a mitochondrial protein) and Claudin 4 (Cldn4) expression were downregulated in the oesophageal epithelium of Nrf2−/− mice, suggesting that energy-dependent tight junction integrity was subject to Nrf2 regulation. ChIP analysis confirmed the binding of Nrf2 to Cldn4 promoter.

Conclusions Nrf2 deficiency impairs oesophageal barrier function through disrupting energy-dependent tight junction.

  • EPITHELIAL BARRIER
  • EPITHELIAL PERMEABILITY
  • OESOPHAGEAL DISEASE
  • OESOPHAGEAL REFLUX
  • OESOPHAGEAL STRICTURES

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