Research Mentor Name

Ahmed Ibrahim

Research Mentor Email Address

ahmed.ibrahim@wayne.edu

Institution / Department

Wayne State University School of Medicine Department of Ophthalmology, Visual, and Anatomical Sciences

Document Type

Research Abstract

Research Type

basicbio

Graduate Level Research

no

Abstract

Background: This study examines how fatty acid synthesis and β-oxidation pathways support the integrity of the human retinal endothelial barrier under normal conditions and identifies key enzymes required for maintaining barrier stability. Understanding these pathways helps address gaps in knowledge about retinal changes in disease and highlights potential pharmacologic targets.

Methods: Human retinal endothelial cells (HRECs) were cultured on electrode arrays and monitored by Electric Cell-Substrate Impedance Sensing (ECIS) until stable membrane resistance was achieved. Media were then replaced with solutions containing published inhibitors of fatty acid synthesis or β-oxidation. Barrier effects were tracked for 80 h. Cytotoxicity (LDH assays) was measured at 12, 24, and 48 h. Inhibitor impact was evaluated using endpoint and area under the curve (AUC) analyses at 50 h. ANOVA with Tukey post hoc tests was performed versus DMSO controls.

Results: Eight inhibitors were tested at 2–3 concentrations each. Cerulenin (FAS; 10, 20 µM), Etomoxir (CPT I; 10–40 µM), and Trimetazidine (3-Ketoacyl-CoA Thiolase; 10–40 µM) decreased barrier resistance dose-dependently without 12 h cytotoxicity. Triacsin C (ACSL; 1 µM) and TOFA (ACC; 10–40 µM) increased resistance. ELOVL-1-22 and ELOVL-1-27 (VLCFA elongase inhibitors) had no effect. Perhexiline (CPT II) was cytotoxic at 10 µM and showed no significant effects at 0.1–1 µM.

Conclusion: β-oxidation appears essential for HREC barrier integrity: inhibition of β-oxidation enzymes reduced resistance, whereas ACC inhibition—which increases β-oxidation—enhanced it. FAS inhibition (raising acetyl-CoA and malonyl-CoA) decreased integrity, while ACSL inhibition (increasing palmitate availability for β-oxidation) improved it.

Disciplines

Medicine and Health Sciences

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