Hormones and Growth FactorsInhibition of oxidant-induced barrier disruption and protein tyrosine phosphorylation in Caco-2 cell monolayers by epidermal growth factor
Section snippets
Cell culture
Caco-2 cells, originally obtained from Dr. Jeffrey Field, were maintained under standard cell culture conditions at 37° in Dulbecco’s modified Eagle’s medium (DMEM) containing 20% (v/v) FBS. Cells were grown on polycarbonate membranes in Transwells (6.5 mm; Costar). Experiments were performed on day 12 or 13 after seeding cells onto Transwells (within passages No. 40–55). Under these conditions, confluent monolayers attained steady-state resistance to passive transepithelial ion flow, and
Paracellular permeability in Caco-2 cell monolayer
Baseline TER of Caco-2 cell monolayers (12- or 13-day post-seeding) varied from 300 to 450 Ω · cm2. Administration of H2O2 to the basal surface at final concentrations of 0.1 to 5.0 mM reduced the TER in a time- and concentration-related manner (Fig. 1 ). Administration of EGF (30 nM) to the apical or basal buffer by itself produced no significant effect on baseline TER. However, EGF administered 60 min prior to H2O2 (5 mM) significantly inhibited the H2O2-induced decrease in TER (Fig. 2A ).
Discussion
The present study demonstrated that EGF delays oxidant-induced increase in paracellular permeability in Caco-2 and T-84 cell monolayers. Exposure to H2O2 (0.1 to 5.0 mM) reduced the TER of Caco-2 cell monolayers in a time- and concentration-related manner. This effect of H2O2 was associated with a decrease in sodium chloride dilution potential and an increase in unidirectional flux of [3H]mannitol. As described above, the concentration of H2O2 used is well within the pathophysiologic
Acknowledgements
This study was supported by a grant from the University Research Committee, Medical University of South Carolina.
References (37)
- et al.
Free radicals and other reactive oxygen metabolites in inflammatory bowel diseaseCause, consequence or epiphenomenon?
Pharmacol Ther
(1992) - et al.
Neutrophils adherent to a nonphagocytosable surface (glomerular basement membrane) produce oxidants only at the site of attachment
Blood
(1985) Isolation of mouse submaxillary gland protein accelerating incisor eruption and eyelid opening in the newborn animal
J Biol Chem
(1962)- et al.
Role of epidermal growth factor and sulfhydryls in stress-induced gastric lesions
Gastroenterology
(1990) - et al.
Role of salivary glands and epidermal growth factor (EGF) in gastric secretion and mucosal integrity in rats exposed to stress
Regul Pept
(1991) - et al.
Epidermal growth factor protects mouse ileal mucosa from Triton X-100-induced injury
Eur J Pharmacol
(1996) - et al.
Interruption of a transforming growth factor α autocrine loop in Caco-2 cells by antisense oligodeoxynucleotides
Gastroenterology
(1995) - et al.
Characterization of the human colon carcinoma cell line (Caco-2) as a model for intestinal epithelial permeability
Gastroenterology
(1989) Biologically active peptides in the gastrointestinal lumen
Life Sci
(1991)- et al.
Epidermal growth factor receptor of the intestinal enterocyte. Localization of lateral basal but not brush border membranes
J Biol Chem
(1989)
Developmental regulation of epidermal growth factor receptor kinase in rat intestine
Gastroenterology
Toxicity, single-strand breaks, and 5-hydroxymethyl-2′-deoxyuridine formation in human breast epithelial cells treated with hydrogen peroxide
Free Radic Biol Med
Genistein, a specific inhibitor of tyrosine-specific protein kinases
J Biol Chem
Thiazolidine-diones. Biochemical and biologic activity of a novel class of tyrosine protein kinase inhibitors
J Biol Chem
Colon absorptive epithelial cells lose proliferative response to TGFα as they differentiate
Exp Cell Res
Oxygen-derived speciesTheir relation to human disease and environmental stress
Environ Health Perspect
Free radicals and inflammationSuperoxide-dependent activation of a neutrophil chemotactic factor in plasma
Proc Natl Acad Sci USA
Oxidant-induced disruption of the intestinal epithelial barrier functionRole of protein tyrosine phosphorylation
Am J Physiol
Cited by (86)
Cell membrane nanomaterials composed of phospholipids and glycoproteins for drug delivery in inflammatory bowel disease: A review
2023, International Journal of Biological MacromoleculesROS-responsive thioketal-linked alginate/chitosan carriers for irritable bowel syndrome with diarrhea therapy
2022, International Journal of Biological MacromoleculesRevaprazan prevented indomethacin-induced intestinal damages by enhancing tight junction related mechanisms
2020, Biochemical PharmacologyTargeting gut barrier dysfunction with phytotherapies: Effective strategy against chronic diseases
2020, Pharmacological ResearchAnti-bacterial susceptibility profiling of Weissella confusa DD_A7 against the multidrug-resistant ESBL-positive E. coli
2019, Microbial PathogenesisCitation Excerpt :In the study, results further ascertained that CFCS has not only shown its potential to interfere with the lipid-protein bilayer but also has the ability to interfere with the DNA integrity [31]. Numerous line of evidence has reported that intracellular ROS is a factor affecting the necessary biomolecules required for cell survival [32]. As assumed, the interacellular ROS levels were increased in all the pathogenic bacteria upon treatment with CFCS, which proves that CFCS is toxic to the tested bacteria, causing maximum cell death.