Effects of EPA on Mouse Endothelial Cell Health

PURPOSE: Cardiovascular disease is the number one killer of adults in the US (~600,000 lives every year: 1 in 4 deaths). Evidence suggests omega-3 fatty acids, such as eicosapentaenoic acid (EPA), commonly found in fish oils, reduce an individuals’ risk for cardiovascular disease. Recent work argues omega-3 fatty acids stimulate an anti-inflammatory response by binding to Free Fatty Acid Receptors (FFAR4). This study evaluated the impact of EPA on endothelial cell health by addressing expression of Cx43 protein (decrease in healthy cells). EPA will decrease Cx43 expression through the FFAR4 (blockade will prevent this decrease). SUBJECTS: Mouse endothelial cells (bEnd.3). METHODS AND MATERIALS: bEnd.3 cells were treated with 30 μM EPA (in ethanol) for 1.5, 3, 6, 12, 24, and 48 hours before protein was isolated (controls were isolated at each time). The protein was loaded equally into a polyacrylamide gel, separated, and transferred to a PVDF membrane and probed for Cx43 (chemiluminescence). The duration of EPA treatment where Cx43 decreased was also treated with the FFAR4 antagonist (AH-7614). ANALYSES: Two sample t-tests compared Cx43 expression ratios (treatment/control) with the value at 1.5 hr (p\u3c0.05). RESULTS: EPA decreased Cx43 expression in bEnd.3 cells at 48 hours. Treatment of these cells for 48 hours with EPA and the FFAR4 blocker (AH-7614 10 uM) is expected to prevent the decrease in Cx43 expression (data in progress). CONCLUSIONS: The health of the endothelium is enhanced by 48 hour EPA treatment that we expect is a result of its binding to the FFAR4