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2017 Cardiovascular Research Day Abstract Book

45 Determination of The

45 Determination of The Effects Of High Fat Feeding On Autotaxin Activity Esias Bedingar 1 • Frederick Onono, PhD 2 1University of Kentucky • 2 Internal Medicine , College of Medicine Undergraduate Obesity is a major public health issue in the United States and is risk factor for several diseases such as coronary artery disease and certain types of cancers. Although the association between obesity and cardiovascular diseases is well known, the underlying molecular mechanisms are still unclear. One of the hypotheses proposed to explain the link between obesity and disease risk is that consumption of high fat diets or increased synthesis and storage of fats is associated with the production of bioactive lipids or lipid-derived molecules that promote disease. Lysophosphatidic acid (LPA) is a family of bioactive lysophospholipids well-recognized as an important signaling molecule acting primarily through actions at cell-surface G-protein coupled receptors. There are two major ways of LPA production. However, the enzyme responsible for making majority of the circulating LPA is autotaxin (ATX), a secreted phospholipase D. Autotaxin is strongly expressed in adipose tissue, thus synthesis of circulating LPA from dietary lipids could be further enhanced in obese individuals. In this study, we investigated the effects of fasting and high-fat feeding on the activity levels of ATX. Healthy volunteer subjects (n=11) were asked to fast overnight. In the morning, their blood was collected at t=0 for plasma preparation. Then, they were given a drink composed of boost protein shake and triglycerides and blood drawn at hourly intervals for up to 8 hours. Autotaxin activity was determined in the plasma using synthetic substrates. Preliminary results indicated that ATX activity is acutely sensitive to fasting and feeding, with the lowest activity observed following fasting and increasing with feeding of a high fat diet. To confirm the specificity of the ATX activity a novel inhibitor was used to inhibit ATX activity. Our findings indicate that ATX is dependent on the feeding status and is increased following feeding with a high fat diet. This study suggests the efficacy of ATX pharmacological inhibition could be enhanced if timed with the onset of feeding. 61

46 Sex-Specific Differences in Cardiac Fibrosis Gregory Milburn 1 • Autumn Conger 1 • Cheavar Blair, PhD 1 • Maya Guglin, MD 2 • Gretchen Wells, MD, PhD 2 • Rebekah Waikel, PhD 3 • Kenneth Campbell, PhD 1 1Physiology, University of Kentucky • 2 Cardiovascular Medicine, University of Kentucky • 3Biological Sciences, Eastern Kentucky University Undergraduate Cardiovascular disease is the leading causes of death in America, which prompted a concerted effort to better understand the causes and to develop innovative therapies. The etiology of heart disease is complex and depends on several factors such as age, race, and sex, but previous research conducted contains gaps, as it often did not look for difference between these groups. This study aims to fill one aspect of this research gap by examining sex-specific differences in cardiac fibrosis in failing and non-failing human hearts. Cardiac fibrosis is a result of cardiac remodeling and causes decreased heart function post cardiac damage. A previous study, conducted in our lab, used Nanostring analysis to examine sex-specific differences in the expression of genes related to fibrosis. The results of this study showed certain gene expressions were specific to sex or had an interaction between the sex and heart failure status. Several of these genes were regulators of collagen, a common type of cardiac fibrosis. We continued this study by quantifying collagen in human heart samples, collected from heart transplants and non-viable organ donors. These samples were stained using an established picrosirius red staining technique, turning the collagen tissue red and normal cardiac tissue yellow. These samples were then quantified using a k-means cluster, via a program written in lab, to eliminate any bias in determining red and yellow tissue. The results are pending as there are still samples left to be completed. We hope to continue pursuing this line of inquiry into sex-specific differences in heart disease by examining passive stiffness, of which collagen is a major component. 62

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