Inhibiting MiR-33a-3p Expression Fails to Enhance ApoAI-Mediated Cholesterol Efflux in Pro-Inflammatory Endothelial Cells

<i>Background and Objectives</i>: Atherosclerosis is an inflammatory condition that results in cholesterol accumulating within vessel wall cells. Atherosclerotic cardiovascular disease is the leading cause of mortality worldwide due to this disease being a major contributor to myocardial...

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Main Authors: Kun Huang, Achala Pokhrel, Jing Echesabal-Chen, Justin Scott, Terri Bruce, Hanjoong Jo, Alexis Stamatikos
Format: Article
Language:English
Published: MDPI AG 2025-02-01
Series:Medicina
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Online Access:https://www.mdpi.com/1648-9144/61/2/329
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author Kun Huang
Achala Pokhrel
Jing Echesabal-Chen
Justin Scott
Terri Bruce
Hanjoong Jo
Alexis Stamatikos
author_facet Kun Huang
Achala Pokhrel
Jing Echesabal-Chen
Justin Scott
Terri Bruce
Hanjoong Jo
Alexis Stamatikos
author_sort Kun Huang
collection DOAJ
description <i>Background and Objectives</i>: Atherosclerosis is an inflammatory condition that results in cholesterol accumulating within vessel wall cells. Atherosclerotic cardiovascular disease is the leading cause of mortality worldwide due to this disease being a major contributor to myocardial infarctions and cerebrovascular accidents. Research suggests that cholesterol accumulation occurring precisely within arterial endothelial cells triggers atherogenesis and exacerbates atherosclerosis. Furthermore, inflamed endothelium acts as a catalyst for atherosclerotic development. Therefore, enhancing cholesterol removal specifically in pro-inflammatory endothelial cells may be a potential treatment option for atherosclerosis. While we have previously shown that inhibiting the microRNA guide strand miR-33a-5p within pro-inflammatory endothelial cells increases both ABCA1 expression and apoAI-mediated cholesterol efflux, it is unknown whether inhibiting the miR-33a-3p passenger strand in pro-inflammatory endothelial cells causes similar atheroprotective effects. In this study, this is what we aimed to test. <i>Materials and Methods</i>: We used plasmid transfection to knockdown miR-33a-3p expression within cultured pro-inflammatory immortalized mouse aortic endothelial cells (iMAECs). We compared ABCA1 expression and apoAI-mediated cholesterol efflux within these cells to cultured pro-inflammatory iMAECs transfected with a control plasmid. <i>Results</i>: The knockdown of miR-33a-3p expression within pro-inflammatory iMAECs resulted in a significant increase in ABCA1 mRNA expression. However, the inhibition of miR-33a-3p did not significantly increase ABCA1 protein expression within pro-inflammatory iMAECs. Moreover, we failed to detect a significant increase in apoAI-mediated cholesterol efflux within pro-inflammatory iMAECs from miR-33a-3p knockdown. <i>Conclusions</i>: Our results indicative that the knockdown of miR-33a-3p alone does not enhance ABCA1-dependent cholesterol efflux within pro-inflammatory endothelial cells. To gain any atheroprotective benefit from inhibiting miR-33a-3p within pro-inflammatory endothelium, additional anti-atherogenic strategies would likely be needed in unison.
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spelling doaj-art-dbe5bfe27f8f4601bedd67ecfce5aefd2025-08-20T03:12:00ZengMDPI AGMedicina1010-660X1648-91442025-02-0161232910.3390/medicina61020329Inhibiting MiR-33a-3p Expression Fails to Enhance ApoAI-Mediated Cholesterol Efflux in Pro-Inflammatory Endothelial CellsKun Huang0Achala Pokhrel1Jing Echesabal-Chen2Justin Scott3Terri Bruce4Hanjoong Jo5Alexis Stamatikos6Department of Food, Nutrition, and Packaging Sciences, Clemson University, Clemson, SC 29634, USADepartment of Food, Nutrition, and Packaging Sciences, Clemson University, Clemson, SC 29634, USADepartment of Food, Nutrition, and Packaging Sciences, Clemson University, Clemson, SC 29634, USAClemson Light Imaging Facility, Clemson University, Clemson, SC 29634, USAClemson Light Imaging Facility, Clemson University, Clemson, SC 29634, USACoulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA 30322, USADepartment of Food, Nutrition, and Packaging Sciences, Clemson University, Clemson, SC 29634, USA<i>Background and Objectives</i>: Atherosclerosis is an inflammatory condition that results in cholesterol accumulating within vessel wall cells. Atherosclerotic cardiovascular disease is the leading cause of mortality worldwide due to this disease being a major contributor to myocardial infarctions and cerebrovascular accidents. Research suggests that cholesterol accumulation occurring precisely within arterial endothelial cells triggers atherogenesis and exacerbates atherosclerosis. Furthermore, inflamed endothelium acts as a catalyst for atherosclerotic development. Therefore, enhancing cholesterol removal specifically in pro-inflammatory endothelial cells may be a potential treatment option for atherosclerosis. While we have previously shown that inhibiting the microRNA guide strand miR-33a-5p within pro-inflammatory endothelial cells increases both ABCA1 expression and apoAI-mediated cholesterol efflux, it is unknown whether inhibiting the miR-33a-3p passenger strand in pro-inflammatory endothelial cells causes similar atheroprotective effects. In this study, this is what we aimed to test. <i>Materials and Methods</i>: We used plasmid transfection to knockdown miR-33a-3p expression within cultured pro-inflammatory immortalized mouse aortic endothelial cells (iMAECs). We compared ABCA1 expression and apoAI-mediated cholesterol efflux within these cells to cultured pro-inflammatory iMAECs transfected with a control plasmid. <i>Results</i>: The knockdown of miR-33a-3p expression within pro-inflammatory iMAECs resulted in a significant increase in ABCA1 mRNA expression. However, the inhibition of miR-33a-3p did not significantly increase ABCA1 protein expression within pro-inflammatory iMAECs. Moreover, we failed to detect a significant increase in apoAI-mediated cholesterol efflux within pro-inflammatory iMAECs from miR-33a-3p knockdown. <i>Conclusions</i>: Our results indicative that the knockdown of miR-33a-3p alone does not enhance ABCA1-dependent cholesterol efflux within pro-inflammatory endothelial cells. To gain any atheroprotective benefit from inhibiting miR-33a-3p within pro-inflammatory endothelium, additional anti-atherogenic strategies would likely be needed in unison.https://www.mdpi.com/1648-9144/61/2/329ABC transportersantagomiRcardiovascular diseaselipopolysaccharidereverse cholesterol transportsystemic inflammation
spellingShingle Kun Huang
Achala Pokhrel
Jing Echesabal-Chen
Justin Scott
Terri Bruce
Hanjoong Jo
Alexis Stamatikos
Inhibiting MiR-33a-3p Expression Fails to Enhance ApoAI-Mediated Cholesterol Efflux in Pro-Inflammatory Endothelial Cells
Medicina
ABC transporters
antagomiR
cardiovascular disease
lipopolysaccharide
reverse cholesterol transport
systemic inflammation
title Inhibiting MiR-33a-3p Expression Fails to Enhance ApoAI-Mediated Cholesterol Efflux in Pro-Inflammatory Endothelial Cells
title_full Inhibiting MiR-33a-3p Expression Fails to Enhance ApoAI-Mediated Cholesterol Efflux in Pro-Inflammatory Endothelial Cells
title_fullStr Inhibiting MiR-33a-3p Expression Fails to Enhance ApoAI-Mediated Cholesterol Efflux in Pro-Inflammatory Endothelial Cells
title_full_unstemmed Inhibiting MiR-33a-3p Expression Fails to Enhance ApoAI-Mediated Cholesterol Efflux in Pro-Inflammatory Endothelial Cells
title_short Inhibiting MiR-33a-3p Expression Fails to Enhance ApoAI-Mediated Cholesterol Efflux in Pro-Inflammatory Endothelial Cells
title_sort inhibiting mir 33a 3p expression fails to enhance apoai mediated cholesterol efflux in pro inflammatory endothelial cells
topic ABC transporters
antagomiR
cardiovascular disease
lipopolysaccharide
reverse cholesterol transport
systemic inflammation
url https://www.mdpi.com/1648-9144/61/2/329
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