Date of Award
12-2024
Document Type
Dissertation
Degree Name
Doctor of Philosophy (PhD)
Department
Food Technology
Committee Chair/Advisor
Alexis Stamatikos
Committee Member
Hannah Kate Wilson
Committee Member
Khaled Abdelaziz
Committee Member
George Cavender
Abstract
Atherosclerosis is the process of cholesterol buildup within arterial cells, which is considered a leading factor that causes certain cardiovascular diseases. Cholesterol accumulation within pro-inflammatory endothelial cells is thought to induce atherogenesis and accelerate atherosclerosis progression. Evidence suggests inhibiting the microRNA miR-33a in arterial cells may promote atheroprotection via increasing ABCA1 protein expression. MiR-33a is processed into two functional strands, miR-33a-5p and miR-33a-3p. The potential atheroprotective effect of inhibiting miR-33a-5p and/or miR-33a-3p precisely in pro-inflammatory endothelial cells is not fully understood, as data is scant regarding what potential atheroprotective impact ABCA1-dependent cholesterol within pro-inflammatory endothelial cells demonstrates, which occurs via miR-33a-5p/3p inhibition. My dissertation project investigates whether inhibiting miR-33a-5p/3p in cultured pro-inflammatory endothelial cells increases ABCA1-dependent cholesterol efflux. Initially, I transfected inflamed immortalized mouse aortic endothelial cells (iMAEC) with either the plasmid pAntimiR33a5p or the control plasmid pScr, and my results showed an increase for ABCA1-dependent cholesterol efflux within inflamed iMAEC transfected with pAntimiR33a5p versus inflamed control iMAEC. This effect was likely due to decreased miR-33a-5p expression occurring within inflamed iMAEC transfected with pAntimiR33a5p. I also transfected pro-inflammatory iMAEC with either pAntimiR33a3p or control plasmid pScr. For these experiments, I failed to observe increased ABCA1-dependent cholesterol efflux within inflamed endothelial cells transfected with pAntimiR33a3p, even though I still did detect a decrease in miR-33a-3p expression and increased ABCA1 mRNA expression in these transfected pro-inflammatory endothelial cells. Thus, inhibition of miR-33a-3p alone does not appear to recapitulate the same atheroprotective properties as inhibiting miR-33a-5p within these cells. Based on my findings, specifically inhibiting miR-33a-5p within the pro-inflammatory endothelium of atherogenic animal models may confer atheroprotection and so future studies should be conducted which assess atheroprotective efficacy of miR-33a-5p inhibition precisely occurring within the pro-inflammatory endothelial cells of atherogenic mice.
Recommended Citation
Huang, Kun, "The Atheroprotective Impact of Anti-miR-33a-5p/3p in Cultured Pro-Inflammatory Endothelial Cells" (2024). All Dissertations. 3805.
https://open.clemson.edu/all_dissertations/3805