Lactobacillus casei and Epidermal Growth Factor Prevent Osmotic Stress-Induced Tight Junction Disruption in Caco-2 Cell Monolayers

• Published in: Cells (Basel, Switzerland) Switzerland), 2021-12, Vol.10 (12), p.3578
• Main Authors: Samak, Geetha, Rao, Rupa, Rao, Radhakrishna
• Format: Article
• Language: English
• Subjects: Actin; Actin Cytoskeleton - drug effects; Actin Cytoskeleton - metabolism; Adherens Junctions - drug effects; Adherens Junctions - metabolism; Antibodies; Binding sites; Caco-2 Cells; Cell culture; Confocal microscopy; Cytoskeleton; EGF; Enzyme inhibitors; Epidermal growth factor; Epidermal Growth Factor - pharmacology; Epithelium; ErbB Receptors - metabolism; Gastrointestinal diseases; Humans; Immunofluorescence; Inflammatory bowel disease; Kinases; Laboratories; Lactobacillus casei; Lactobacillus casei - physiology; MAP kinase; Mitogen-Activated Protein Kinases - metabolism; Osmosis; Osmotic Pressure - drug effects; Osmotic stress; Permeability; probiotic; Probiotics; Protein kinase C; Protein Kinase C - metabolism; Protein-tyrosine kinase; Proteins; tight junction; Tight Junctions - drug effects; Tight Junctions - metabolism; Tight Junctions - pathology
• ISSN: 2073-4409; 2073-4409
• DOI: 10.3390/cells10123578

Osmotic stress plays a crucial role in the pathogenesis of many gastrointestinal diseases. and epidermal growth factor (EGF) effects on the osmotic stress-induced epithelial junctional disruption and barrier dysfunction were investigated. Caco-2 cell monolayers were exposed to osmotic stress in the presence or absence of or EGF, and the barrier function was evaluated by measuring inulin permeability. Tight junction (TJ) and adherens junction integrity were assessed by immunofluorescence confocal microscopy. The role of signaling molecules in the and EGF effects was determined by using selective inhibitors. Data show that pretreatment of cell monolayers with or EGF attenuates osmotic stress-induced TJ and adherens junction disruption and barrier dysfunction. EGF also blocked osmotic stress-induced actin cytoskeleton remodeling. U0126 (MEK1/2 inhibitor), the MAP kinase inhibitor, blocked EGF-mediated epithelial protection from osmotic stress. In contrast, the -mediated epithelial protection from osmotic stress was unaffected by U0126, AG1478 (EGFR tyrosine kinase inhibitor), SP600125 (JNK1/2 inhibitor), or SB202190 (P38 MAP kinase inhibitor). On the other hand, Ro-32-0432 (PKC inhibitor) blocked the -mediated prevention of osmotic stress-induced TJ disruption and barrier dysfunction. The combination of EGF and is more potent in protecting the barrier function from osmotic stress. These findings suggest that and EGF ameliorate osmotic stress-induced disruption of apical junctional complexes and barrier dysfunction in the intestinal epithelium by distinct signaling mechanisms.