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Kerstin Birke, Friedrich E. Kruse, Ulrich-Christoph Welge-Lüßen, Marco T. Birke; Effects Of Omega-3 And Omega-6 Fatty Acids On H2O2 Stimulated NFkB Activation And Secretion Of Interleukins 6 And 8 In Primary Human Trabecular Meshwork Cells. Invest. Ophthalmol. Vis. Sci. 2012;53(14):3229.
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To analyze effects of Omega-3 and Omega-6 fatty acids on NFΚB, IL-6 and IL-8 production, and to test their antagonizing capacities against H2O2 stimulation of these factors in primary human TM cells.
Primary hTM cells were derived from 5 different donors and propagated in Ham’s F10 containing 10% FCS and 1% penicillin/streptomycin mixture to passage 5 at maximum. Treatment was performed in serum-free F10 supplemented to 50µM Omega-3 or 16µM Omega-6 for 48 hours in triplicates. A second set of hTM cells was treated analogously but additionally exposed to 300µM H2O2 for 1h to induce cellular oxidative stress. Untreated hTM cells served as controls for both approaches. The mRNA expression of Il-6, Il-8, Il-1a and ELAM-1 was analyzed by realtime-PCR. Contents of IL-6 and IL-8 in the medium were quantified by ELISA. Amounts of NFΚB were determined by a NoShift™ NFΚB Assay on nuclear protein extracts of hTM cells.
Realtime PCR data indicate that Omega-6 and Omega-3 both were able to reduce H2O2 mediated induction of IL-1α, IL-6, IL-8, NFΚB and ELAM-1 mRNA. The extents of repressions were different, however. On the protein level, ELISA assays suggest that both fatty acids counteract IL-6 stimulation, whereas IL-8 stimulation appeared to be efficiently repressed by Omega-3 only. A similar result was achieved in the NFΚB assay, indicating H2O2 counteracting effects for Omega-3 only.
Both fatty acids appeared to act similarily on mRNAs of typical oxidative stress markers. On protein level, however, Omega-3 seems to be more effective, thus proposing Omega-3 to be more beneficial with the respect to prevention from oxidative stress.
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