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Bioactivity of the endocannabinoid arachidonoylethanolamide in cultured bovine endothelial cells.

C. C. F. Walker

Abstract:

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Bioactivity of the endocannabinoid arachidonoylethanolamide in cultured bovine endothelial cells.
C. C. F. Walker*, L. M. Sordillo. Michigan State University East Lansing, MI.

Vascular endothelial cells are crucial inflammatory-mediating cells susceptible to compromised barrier integrity during coliform mastitis. Endocannabinoid arachidonoylethanolamide (AEA) is a lipid mediator that can affect barrier integrity through modulation of network formation, proliferation, and viability in human and rodent endothelial cells during endotoxin challenge. We hypothesized that the endocannabinoid AEA will increase barrier integrity of primary cultured bovine aortic endothelial cells (BAEC) challenged with lipopolysaccharide (LPS). Cells were treated with varying AEA concentrations (10 nM to 5 �M) for up to 12 h following pre-treatment with 25 ng/mL LPS. Endothelial barrier integrity was continuously assessed by recording electrical resistance using an electric cell-substrate impedance sensing system and reported resistance was normalized to a media control. Cell proliferation and viability were assessed using commercially available plate-based assays. All AEA treatments were compared with LPS control group and analyzed using the ANOVA procedure in SAS9.4. Physiological-relevant concentrations (<1 �M) increased proliferation and viability of BAEC by up to 21.5 � 2.283% and 19.37 � 2.564%, respectively, at 1 and 2 h post-treatment (P < 0.05). Normalized electrical resistance was increased for 2 h after AEA treatment at 100 nM (0.105 � 0.0137 Ω) and 500 nM (0.123 � 0.015 Ω) (P < 0.05). No differences in resistance were noted for any other treatments <1 �M (P > 0.05). Doses >1 �M decreased electrical resistance, proliferation, and viability for up to 2 h post-treatment (P < 0.05). All treatment effects were lost at 6 or 12 h (P > 0.05). Physiological AEA concentrations of <1 μM improve barrier integrity of BAEC when challenged with LPS, in contrast to the potential toxicity associated with super-physiological concentrations of >1 �M. Ongoing projects are focused on the mechanistic effects of AEA on endothelial cell barrier integrity with the goal of reducing the severity of coliform mastitis.

Keywords: coliform mastitis, endocannabinoids, endothelial cells.

Biography: Second year PhD student in the Meadow Brook Endowed Immunobiology Laboratory at Michigan State. Focus of research is the bovine endocannabinoid system and its implications on endothelial cell health under endotoxin challenge. Before starting PhD, worked as a nutrition consultant and operations manager for a human grade cricket farm.