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Abstract zur Publikation: Stimulus (polyphenol, IFN-γ, LPS)-dependent nitric oxide production and antileishmanial effects in RAW 264.7 macrophages

Kolodziej H, Radtke OA, Kiderlen AF (2008): Stimulus (polyphenol, IFN-γ, LPS)-dependent nitric oxide production and antileishmanial effects in RAW 264.7 macrophages
Phytochem. 69: 3103-3110.

The effects of interferon (IFN-γ), lipopolysaccharide (LPS), and some polyphenols as individual stimuli, as well as in various combinations on NO production in non-infected and infected macrophage-like RAW 264.7 cells were investigated, with emphasis on the NO/parasite relationship. In non-infected and in Leishmania parasitized cells, gallic acid significantly inhibited the IFN-γ- and LPS-induced NO detected in the supernatant. This effect was less prominent in IFN-γ- than in LPS-stimulated cells. Interestingly, and in contrast to non-infected cells, gallic acid inhibited NO production only when added within 3 h after IFN-γ + LPS. Addition of gallic acid following prolonged (24 h) incubation periods with IFN-γ + LPS no longer inhibited, sometimes even enhanced NO release. Notably, an excellent NO/parasite kill relationship was evident from all experiments. This study was extended to a series of polyphenols (3-O-shikimic acid, its 3,5-digalloylated analogue, catechin, EGCG, and a procyanidin hexamer) with proven immunostimulatory activities. Although these compounds themselves were found to be only weak NO-inducers, the viability of intracellular Leishmania parasites was considerably reduced. Furthermore, their dose-dependent effects on macrophages NO release was determined in the presence of IFN-γ and/or LPS. Again, non-infected and infected cells differed significantly in the NO response, while inhibition of IFN-γ and/or LPS-induced NO production by the tested polyphenols strongly depended on the given time of exposure and the sequence of immunological stimuli. A strong inverse correlation between NO levels and intracellular survival rates of Leishmania parasites supported the assumption that the observed inhibition of NO was not simply due to interference with the Griess assay used for detection.

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