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EGF and L-Glutamine Accelerates the Restoration of Epithelial Barrier Function From Oxidative Stress-Induced Injury. Nichole Barnhart Biology/Microbiology TTU. Project Summary.
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EGF and L-Glutamine Accelerates the Restoration of Epithelial Barrier Function From Oxidative Stress-Induced Injury Nichole Barnhart Biology/Microbiology TTU
Project Summary • Introduction: TJ form a barrier to the diffusion of toxins, allergens and pathogens from the intestinal lumen into the tissues. Oxidative stress occurs when there is an accumulation of free radicals, such as superoxide, hydrogen peroxide and hydroxyl radical, which have one or more unpaired electrons, and can damage cells, by oxidizing proteins, fats and DNA.
Problem and Objective • What effects do EGF and L-glutamine have on the epithelial barrier function of intestinal cells? • Objective: The objective is to see if EGF and L-glutamine accelerate the restoration of epithelial barrier function from oxidative stress-induced injury.
Method and Hypothesis • Method: Caco-2 cells grown on Transwell inserts. Induce oxidative stress with hydrogen peroxide. Restoration using EGF and L-glutamine. Paracellular permeability evaluated using transepithelial electrical resistance (TER) and FITC-labeled inulin. Stain for TJ proteins by immunofluorescence. • Hypothesis: EGF and L-glutamine accelerate the restoration of epithelial barrier function from oxidative stress-induced injury.
Literary Sources Studies have shown that: • Tight junctions of cells regulate paracellular permeability (Anderson 1995) • Free radicals such as superoxide and hydrogen peroxide cause tissue injury and injury to TJ (Del Maestro 1980) 3. EGF inhibits the barrier disfunction of Caco-2 cell monolayers (Rao 1999)
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Expected Results • Expected Results: I anticipate that EGF and L-glutamine will accelerate the restoration of epithelial barrier function from oxidative stress-induced injury. • Advantages: There are many clinical syndromes and diseases associated with the disruption of TJ and increased intestinal permeability and the recovery time from these diseases can be lowered significantly.
Project Timeline and Budget • This project was done during the summer of 2004 and UTHSC with the help of Dr. RK Rao and others of the Physiology Dept. • The budget of this research project was covered by grants received by the UTHSC Physiology Dept.