ISOPROSTANES MITIGATE OXIDATIVE STRESS IN BOVINE VASCULAR ENDOTHELIUM DURING INFLAMMATION

<b>Project Methods</b><br> An in vitro model will be developed utilizing primary bovine aortic and mammary endothelial cells (representing systemic and local inflammatory responses, respectively)from 6 Holstein dairy cows. Two omega-6 isoprostanes will be utilized: 15-F2t-IsoP, which is the most widely cited isoprostane in the literature, and 8-iso-PGE2. The omega-3-derived 15-F3t-IsoP will also be utilized. Accomplishing the aims will involve determiningwhich concentrations and the timing in whichisoprostanes will demonstrate the most potency in mitigating oxidative stress. Outcomes of oxidative stress, including ROS production, cell viability, apoptosis, and endothelial cell barrier intergrity, will be evaluated via several techniques. The aforementioned model, with modifications,will be utilizedto determine if isoprostanes can be modulated to benefit the vasculature. In brief, bovine endothelial cells will be supplemented with omega-6- and -3 fatty acids and various micronutrients with antioxidant capacity (e.g., selenium, vitamin E, vitamin A)to test the working hypothesis thesupplementation ofcell culturewill result in improved endothelial cellfunction. Statistics will be run with a one-way ANOVA with Tukey's Adjustment for normally distributed data or a Kruskal-Wallis H test with Dunn's post hoc test for non-normal data in SAS. When appropriate, a two-way ANOVA will be performed. Statistical significance will be set atP< 0.05.The data will be disseminated via seminars and manuscripts published in peer-reviewed journals. Evaluations will include successfuldevelopment of an in vitro model to test the hypothesis that isoprostanes can mitigate oxidative stress during acute inflammation in cattle.