Background and Aim: High intakes of -3 fatty acids have been associated with systemic anti-inflammatory effects and cardiovascular protection, but the molecular basis responsible for these effects remains incompletely defined. Using a DNA microarray technology we investigated the early gene expression profile of human vascular endothelium conditioned by DHA under proinflammatory conditions. Materials and Methods: HUVEC were treated with 50 μmol/L DHA for 48 hours and then stimulated with 5 ng/mL IL-1 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 profile 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 and Conclusions: IL-1 stimulation significantly changed the expression of 1474 genes: 815 resulted down -regulated while 659 resulted up-regulated. Focusing on the 659 IL-1-upregulated genes, we observed that DHA significantly attenuated the expression of 88 such genes. The application of the Ingenuity pathway analysis software allowed us to pinpoint immunological-, inflammatory and atherogenic-related pathways as the most affected. In particular, we identified new target molecules involved in atherosclerosis, including tubulin beta polypeptide[TUBB]2 A and phosphodiesterase PDE5A; and in angiogenesis, including transforming growth factor [TGF]- 2 and chemokine (C-X-C motif) ligand 10. In conclusion, DHA widely affects endothelial gene expression; the identification of novel genes susceptible to DHA will certainly improve our understanding of mechanisms by which fish oils may prevent or attenuate human chronic diseases including atherosclerosis
Microarray analysis of human umbilical vein endothelial cells (HUVEC) treated with the omega-3 fatty acid docosahexaenoic acid (DHA) highlights new anti-atherosclerotic and anti angiogenic properties for fish and fish oil.
MARTINELLI, ROSANNA;
2010-01-01
Abstract
Background and Aim: High intakes of -3 fatty acids have been associated with systemic anti-inflammatory effects and cardiovascular protection, but the molecular basis responsible for these effects remains incompletely defined. Using a DNA microarray technology we investigated the early gene expression profile of human vascular endothelium conditioned by DHA under proinflammatory conditions. Materials and Methods: HUVEC were treated with 50 μmol/L DHA for 48 hours and then stimulated with 5 ng/mL IL-1 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 profile 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 and Conclusions: IL-1 stimulation significantly changed the expression of 1474 genes: 815 resulted down -regulated while 659 resulted up-regulated. Focusing on the 659 IL-1-upregulated genes, we observed that DHA significantly attenuated the expression of 88 such genes. The application of the Ingenuity pathway analysis software allowed us to pinpoint immunological-, inflammatory and atherogenic-related pathways as the most affected. In particular, we identified new target molecules involved in atherosclerosis, including tubulin beta polypeptide[TUBB]2 A and phosphodiesterase PDE5A; and in angiogenesis, including transforming growth factor [TGF]- 2 and chemokine (C-X-C motif) ligand 10. In conclusion, DHA widely affects endothelial gene expression; the identification of novel genes susceptible to DHA will certainly improve our understanding of mechanisms by which fish oils may prevent or attenuate human chronic diseases including atherosclerosisI documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
