Dysregulation of toll-like receptor (TLR) signaling within the gastrointestinal epithelium
has been associated with uncontrolled inflammation and tumorigenesis. We sought to
evaluate the role of TLR4 in the development of gastroesophageal reflux-mediated inflammation
and mucosal changes of the distal esophagus. Verified human esophageal Barrett's cells
with high grade dysplasia (CPB) and esophageal adenocarcinoma cells (OE33) were treated
with deoxycholic acid for 24 hours. Cells were pretreated with a TLR4-specific inhibitor
peptide 2 hours prior to deoxycholic acid treatment. Inflammatory markers were evaluated
using immunoblotting and enzyme-linked immunosorbent assay. A surgical reflux mouse
model was generated by performing a side-to-side anastomosis between the second portion
of the duodenum and the gastroesophageal junction. Control animals underwent laparotomy
with incision and closure of the esophagus superior to the gastroesophageal junction
(sham procedure). Esophageal sections were evaluated using hematoxylin and eosin staining
and immunohistochemistry. Deoxycholic acid increased expression of inflammatory markers
including intercellular adhesion molecule-1, vascular cell adhesion molecule-1, and
interleukin 8. Pretreatment with a TLR4 inhibitor significantly decreased deoxycholic
acid-induced inflammatory marker expression. C3H/HeNCrl mice demonstrated a significant
increase in mucosal hyperplasia and proliferation following DGEA compared to sham
procedure. TLR4 mutant mice (C3H/HeJ) undergoing DGEA demonstrated an attenuated hyperplastic
and proliferative response compared to C3H/HeNCrl mice. Inhibition of TLR4 signaling
attenuates reflux-induced inflammation in vivo. These findings identify TLR4 inhibition
as a potential therapeutic target to halt the progression of pathologic esophageal
changes developing in the setting of chronic gastroesophageal reflux disease.
Graphical abstract
Graphical Abstract
Keywords
Abbreviations:
DCA (Deoxycholic acid), DGEA (Duodeno-gastroesophageal anastomosis), EAC (Esophageal adenocarcinoma), ELISA (Enzyme-linked immunosorbent assay), FBS (Fetal bovine serum), GAPDH (Glyceraldehyde-3-phosphate dehydrogenase), H&E (Hematoxylin and eosin), ICAM-1 (Intercellular adhesion molecule-1), IL-8 (Interleukin 8), LPS (Lipopolysaccharide), PBS (Phosphate-buffered saline), TLR (Toll-like receptor), TLR4 (Toll-like receptor 4), VCAM-1 (Vascular cell adhesion molecule-1)To read this article in full you will need to make a payment
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Article info
Publication history
Published online: September 14, 2021
Footnotes
Institutional Review Board Approval: Our animal protocol was approved by the Institutional Animal Care and Use Committee at the University of Colorado Anschutz Medical Campus (protocol #00893, approved 4/16/2019).
Funding: This work was funded by the University of Colorado School of Medicine Department of Surgery, Division of Cardiothoracic Surgery.
Conflicts of Interest: The authors have no conflicts of interest to disclose.
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