High intakes of omega-3 fatty acids have been associated with systemic anti-inflammatory effects and cardiovascular protection, but the molecular basis for these effects remains incompletely defined. Using a DNA microarray technology we investigated the early gene expression profile of human vascular endothelial cells conditioned by DHA under proinflammatory conditions. Methods. Human umbilical vein endothelial cells (HUVEC) were treated with 50 µmol/L DHA for 48 hours and then stimulated with 5 ng/mL IL-1beta for 3 hours. Total RNA was extracted, and qualitatively and quantitatively analyzed with a NanoDrop Spectrophotometer and an Agilent Bioanalyzer before RNA labeling and purification. Gene expression profiling was performed with an Agilent Whole Human Genome Oligo Microarray covering 41 000 unique genes and transcripts. Slides were scanned with the Agilent’s scanner and images processed using Agilent Feature Extraction software. The raw data were further processed with the GeneSpring® 10 software and differentially expressed RNA identified using Benjamini and Hochberg False Discovery Rate with a p-value <0.05. Functional and network analyses were identified by the Ingenuity Pathways version 8.0 Analysis. Results IL-1 stimulation significantly changed the expression of 1474 genes: 815 had decreased, while 659 had increased. Out of the 659 IL-1-upregulated genes, DHA significantly attenuated the expression of 88 genes. The Ingenuity pathway analysis software indicated immunological-, inflammatory- and atherogenic pathways as the most affected. In particular, we identified new target molecules involved in atherosclerosis, including tubulin beta polypeptide[TUBB]2 A and phosphodiesterase[PDE]5A; and in angiogenesis, including transforming growth factor[TGF]-beta 2 and chemokine (C-X-C motif) ligand 10. Conclusions: DHA widely affects endothelial gene expression; the identification of novel genes susceptible to regulation by DHA will certainly improve our understanding of mechanisms by which omega-3 fatty acids may prevent or attenuate human chronic diseases including atherosclerosis.
Microarray analysis of human umbilical vein endothelial cells treated with the omega-3 fatty acid docosahexaenoic acid (DHA) highlights new anti-atherosclerotic and anti-angiogenic properties for fish
MARTINELLI, ROSANNA;
2011-01-01
Abstract
High intakes of omega-3 fatty acids have been associated with systemic anti-inflammatory effects and cardiovascular protection, but the molecular basis for these effects remains incompletely defined. Using a DNA microarray technology we investigated the early gene expression profile of human vascular endothelial cells conditioned by DHA under proinflammatory conditions. Methods. Human umbilical vein endothelial cells (HUVEC) were treated with 50 µmol/L DHA for 48 hours and then stimulated with 5 ng/mL IL-1beta for 3 hours. Total RNA was extracted, and qualitatively and quantitatively analyzed with a NanoDrop Spectrophotometer and an Agilent Bioanalyzer before RNA labeling and purification. Gene expression profiling was performed with an Agilent Whole Human Genome Oligo Microarray covering 41 000 unique genes and transcripts. Slides were scanned with the Agilent’s scanner and images processed using Agilent Feature Extraction software. The raw data were further processed with the GeneSpring® 10 software and differentially expressed RNA identified using Benjamini and Hochberg False Discovery Rate with a p-value <0.05. Functional and network analyses were identified by the Ingenuity Pathways version 8.0 Analysis. Results IL-1 stimulation significantly changed the expression of 1474 genes: 815 had decreased, while 659 had increased. Out of the 659 IL-1-upregulated genes, DHA significantly attenuated the expression of 88 genes. The Ingenuity pathway analysis software indicated immunological-, inflammatory- and atherogenic pathways as the most affected. In particular, we identified new target molecules involved in atherosclerosis, including tubulin beta polypeptide[TUBB]2 A and phosphodiesterase[PDE]5A; and in angiogenesis, including transforming growth factor[TGF]-beta 2 and chemokine (C-X-C motif) ligand 10. Conclusions: DHA widely affects endothelial gene expression; the identification of novel genes susceptible to regulation by DHA will certainly improve our understanding of mechanisms by which omega-3 fatty acids may prevent or attenuate human chronic diseases including atherosclerosis.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
