Abstract 3048: The Role Of 17:0(2r-oh) Ceramide In Lps Activated Macrophages

Abstract

Macrophages play a crucial role in the initiation and progression of atherosclerosis, contributing to the buildup of fatty deposits which results in the plaque formation in the walls of arteries. Macrophages modulate the mediators of inflammation and immune response in atherosclerosis. Bacterial lipopolysaccharide (LPS) exerts significant effects on these macrophages including secretion of pro-inflammatory cytokines. The current study is designed to examine the role of sphingosine intermediate N-(2'-(R)-hydroxyheptadecanoyl)-D-erythro-sphingosine (17:0(2R-OH) in the macrophage’s inflammation potential. Couple of studies have indicated a protective role for sphingosine however 17:0(2R-OH) didn’t receive adequate attention. Study design: Mouse J774 macrophages were cultured in Dulbecco’s Modified Eagle’s medium with 4.5g/L glucose and sodium pyruvate without L-glutamine containing 10% fetal bovine serum and 1% penicillin/streptomycin in a humidified atmosphere with 5% CO 2 at 37 °C. The cells were treated in 6-well plates at a density of 5.5х10 6 cells/well. Sphingosine was dissolved in DMSO prior to the treatment. The cells were treated with the 17:0(2R-OH) with 3 different concentrations (5, 25, 50 μg) in duplicates with or without LPS except for the controls. The cells and media were collected at 3 time points (12,24,36) hours. Cells were harvested in Trizol reagent, RNA was then extracted from cells and genes expression were determined in RTPCR using SYBR Green. Results and Conclusion: Our initial microscopic observations suggest that 17:0(2R-OH) potentially modulates inflammation in macrophages, this appears to be dose-dependent. 17:0(2R-OH) has been identified in marine and freshwater fish, marine oils, seaweed, sea urchins, sea cucumbers and may have dietary benefits.