Presentation Type

Poster Presentation

Abstract

Human Defensin 5 (HD5) is an antimicrobial peptide present in the small intestine that plays a role in gut homeostasis. We have been investigating the role of HD5 in colon cells, but research shows that HD5 levels in the small and large intestine vary significantly in Crohn's. Previous data from our lab suggests that HD5 leads to a decrease in wound healing and an increase in apoptosis in colon cells, we want to see if the same is true in the small intestine. So, we are investigating the effect of HD5 on expression of β-catenin, a key protein in the Wnt signaling pathway, in IEC-6 rat small intestinal epithelial cells. We hypothesize that HD5 promotes wound healing and enhances β-catenin expression in IEC-6 cells. To test this, we cultured IEC-6 cells and performed a wound closure assay to compare wound healing between the vehicle control and experimental groups - 0.5 ug/mL, 1.0 ug/mL, and 2.0 ug/mL of HD5. We then extracted protein and RNA to perform a Western Blot and q-RT-PCR for expression analysis. The rate of wound closure between both treatment groups could not be observed. We saw an increase in β-catenin in HD5-treated cells on our western blot (though not statistically significant) and expect a positive fold change for β-catenin in our q-RT-PCR experiment. Future studies will focus on investigating β-catenin expression in colonic epithelial cells treated with HD5 as a possible mechanism for decreased wound closure in Crohn's Colitis.

Faculty Mentor

Dr. Amanda D. Williams

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Levels of Beta Catenin with Treatment of HD5 in IEC-6 Rat Cells

Human Defensin 5 (HD5) is an antimicrobial peptide present in the small intestine that plays a role in gut homeostasis. We have been investigating the role of HD5 in colon cells, but research shows that HD5 levels in the small and large intestine vary significantly in Crohn's. Previous data from our lab suggests that HD5 leads to a decrease in wound healing and an increase in apoptosis in colon cells, we want to see if the same is true in the small intestine. So, we are investigating the effect of HD5 on expression of β-catenin, a key protein in the Wnt signaling pathway, in IEC-6 rat small intestinal epithelial cells. We hypothesize that HD5 promotes wound healing and enhances β-catenin expression in IEC-6 cells. To test this, we cultured IEC-6 cells and performed a wound closure assay to compare wound healing between the vehicle control and experimental groups - 0.5 ug/mL, 1.0 ug/mL, and 2.0 ug/mL of HD5. We then extracted protein and RNA to perform a Western Blot and q-RT-PCR for expression analysis. The rate of wound closure between both treatment groups could not be observed. We saw an increase in β-catenin in HD5-treated cells on our western blot (though not statistically significant) and expect a positive fold change for β-catenin in our q-RT-PCR experiment. Future studies will focus on investigating β-catenin expression in colonic epithelial cells treated with HD5 as a possible mechanism for decreased wound closure in Crohn's Colitis.

 

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