(VP025) COMBINATION OF EICOSAPENTAENOIC ACID (EPA) AND A STATIN INCREASED EXPRESSION OF DETOXIFICATION PROTEINS IN VASCULAR ENDOTHELIAL CELLS DURING INFLAMMATION

Background: Endothelial cell (EC) dysfunction results from reactive oxygen species (ROS) produced during atherosclerosis. Detoxification enzymes like peroxiredoxins (PRDX), heme oxygenase (HO-1) and glutathione reductase reduce levels of ROS. The omega-3 fatty acid eicosapentaenoic acid (EPA), administered as icosapent ethyl (IPE) improves reduced cardiovascular (CV) event in a manner predicted by circulating EPA levels. We measured the separate versus combined effects of EPA and rosuvastatin on expression of detoxification proteins in vascular ECs under conditions of inflammation caused by the effector of the renin angiotensin system, angiotensin II (Ang II).

METHODS AND RESULTS: Human umbilical vein endothelial cells (HUVECs) were challenged with Ang II (100 nM) for 2 h, then treated with rosuvastatin (10 µM) and/or EPA (40 µM) for 24 h. Global proteomic analysis performed using LC/MS measured relative expression levels simultaneously. Significant (p < 0.05) changes in expression between treatment groups >1-fold were analyzed by differential enrichment analysis of proteomics data (DEP) and included in gene set enrichment analyses (GSEA). Levels of HO-1 were measured using immunochemical approaches and normalized to total protein content. Results showed that rosuvastatin alone did not significantly affect expression of proteins related to ROS detoxification. When combined with EPA, however, rosuvastatin significantly modulated 9 proteins within the “cellular response to reactive oxygen species” pathway (GO: 0034614, pathway adjusted p = 0.0142) compared to Ang II. Among the proteins that were increased with the combination were PRDX5 (1.1-fold, p = 0.037), heat shock protein 70 (1.1-fold, p = 0.028), and glutathione reductase (1.1-fold, p = 0.042). Separately, EPA alone also modulated expression of 9 proteins in this pathway (adjusted-p value = 0.002), five of which were shared with the combination treatment, including thioredoxin reductase 1 p = 0.019). Addition of EPA to rosuvastatin increased HO-1 expression by 173% (p < 0.01) while rosuvastatin separately had no effect.

Conclusion: Combination of a high intensity statin and EPA significantly modulated the cellular response to oxidative stress pathway and detoxification proteins, including peroxiredoxin isoforms, HO-1 and glutathione reductase. The addition of EPA to a statin increased expression of detoxification proteins during inflammation that may contribute to reduced vascular and atherothrombotic risk.