ctivator inhibitor 1 mRNA expression was significantly increased by 2.560.5 fold in v-6 15182727” supplemented cells, whereas v-3 reduced the expression level compared to the controls. On the protein level, in contrast, PAI-1 was significantly increased to 1.560.1 fold in v-3 supplemented cells, while v-6 did not affect PAI-1 protein levels. Exposure to H2O2 stimulated PAI-1 mRNA expression in the controls by 5.660.2 fold and cellular PAI-1 protein levels were increased to 1.460.2 fold. In cells pre-treated with v-6 fatty acid the PAI-1 mRNA was not further increased by H2O2. The same applied for the PAI-1 protein levels, which remained constant after H2O2 stimulation. In cells preincubated with v-3 the PAI-1 mRNA expression even dropped to 0.0560.02 fold after H2O2 addition, protein levels, however, were in the same range than in their corresponding controls. Connective tissue MedChemExpress BIRB-796 growth factor mRNA expression was significantly upregulated in cells supplemented with fatty acids compared to the controls. The v-6 mediated activation 6 February 2012 | Volume 7 | Issue 2 | e31340 Prevention of H2O2 Mediated Changes by v-3/ -6 FA of CTGF was also observed on the protein level by a 1.460.05 fold increase. Protein levels in v-3 treated cells were not increased. H2O2 addition induced expression of CTGF mRNA by 21.263.1 fold in the controls. In v-6 pre-treated cells, H2O2 further increased mRNA expression to 23.768.6 fold, which was not observed in v-3 pre-treated cells. On the protein level, H2O2 resulted in a weak, but significant upregulation of CTGF in the controls. In v-6 pre-treated cells, H2O2 exposure led to a reduced protein level than in the corresponding controls. In v-3 pre-treated cells, H2O2 exposure had no effect on CTGF protein level. Expression of Interleukins and NFkB Expression of IL-1a mRNA was significantly stimulated by v-6, whereas v-3 had no effect. Exposition to H2O2 doubled IL-1a mRNA expression. Neither v-3, nor v-6 could significantly inhibited this stimulation. Expression in H2O2/v-6 double treated cells appeared lower than in v-6 supplemented hTM, differences, however, did not reach the level of statistical significance. IL-6 mRNA expression was not altered by both fatty acids. H2O2 stimulation increased 
IL-6 mRNA to 3.461.4 fold, which could not be impeded by both fatty acids. ELISA analysis of the medium revealed that v-6 stimulated IL-6 secretion which was not observed upon v-3 supplementation. H2O2 lead to a 1.660.3 fold induction of IL-6 protein. Both fatty acids appeared to reduce this ” stimulation, however, only the effect of v-3 was significant. IL-8 mRNA expression appeared to be slightly activated by both fatty acids. However, this was statistically significant for v-3 only. Incubation with H2O2 increased IL-8 synthesis to 4.861.7 fold, which could be counteracted by v-3 supplementation. Addition of v-6 was inefficient here. Analysis of IL-8 in the cells medium indicated that fatty acids alone had no effect on IL-8 February 2012 | Volume 7 | Issue 2 | e31340 Prevention of H2O2 Mediated Changes by v-3/ -6 FA contents. Exposition to H2O2 led to a 2.060.2 fold increase of IL-8, which was statistically significant. With respect to H2O2 counteraction, both fatty acids were ineffective. Realtime PCR analysis of NFkB expression did not reveal any significant influences of both fatty acids on this transcription factor. Data suggested that H2O2 slightly stimulated NFkB, but statistical significance was not reached. Notably, such
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