Honors Theses

Date of Award

Spring 5-6-2022

Document Type

Undergraduate Thesis

First Advisor

James Stewart

Second Advisor

Courtney Roper

Third Advisor

John Bentley

Relational Format

eGrove

Abstract

Diabetes is one of the leading causes of death in the United States, taking more than 90,000 lives annually. Type 2 diabetes mellitus-induced cardiac fibrosis is a common condition seen in diabetic patients. The AGE/RAGE signaling cascade has been strongly associated with this supposed cardiac fibrosis along with hyperactive myofibroblasts which excessively remodel extracellular matrix tissue, leading to damaged and enlarged heart muscle tissue. In diabetic patients, Advanced Glycation End Products (AGEs) are produced in abundance and accumulate in the extracellular matrix of heart tissue. To investigate the role of of the AGE ligand on fibroblast behavior, increasing amounts of commercially available exogenous AGE were introduced as a gradient to compare fibroblast cells of four genotypes: diabetic (db), diabetic RAGE knockout (db RKO), Rap1a (Rap1a), Rap1a knockout (RAPKO). Once the fibroblasts were grown to maturity, they were plated in either high or low glucose growth media. The cell’s migration rates were analyzed by scratching the cultured cells in the center of each well at 0 hours after ligand AGE was applied. Using computer digital editing to mark their movement between 0 and 24 hours across the scratched area after the ligand AGE was applied to the plate of cells, the number of cells crossing or touching the drawn boundaries were then counted. Results from this study estimated trends that could have significance on the effect of hyperglycemia and AGE exposure. Additionally, changes in migration were seen to be more dependent upon genotype rather than exposure conditions.

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