In this study, we focused on comparing the effects of serotonin and its metabolites on the functions of RAW264.7 cells (emphasis on oxidative burst and production of nitric oxide and cytokines), thereby expanding the scope of existing knowledge with advent of novel findings in this field. Changes in production of reactive oxygen species (ROS) by RAW264.7 cells after treatment with serotonin, N-acetylserotonin and melatonin were determined using the chemiluminescence (CL) assay. To exclude the direct scavenging effects of the studied compounds on the CL response, the antioxidant properties of all respective compounds were measured using TRAP and amperometrical method. Nitric oxide (NO) production was measured by Griess reagent and inducible NO synthase (iNOS) expression by Western blot. Cytokine production was assessed using the Mouse Cytokine Panel A Array kit and ELISA. We showed that all tested compounds were able to reduce oxidative stress, as well as inhibit production of inflammatory cytokines by macrophages. Of the tested compounds, serotonin and N-acetylserotonin were markedly better antioxidants than melatonin. In comparison, other effects of tested compounds were very similar. It can be concluded that antioxidant capacity of tested compounds is a major advantage in the early stages of inflammation. Since plasma concentrations of N-acetylserotonin and melatonin are lower than serotonin, it can be deduced that serotonin plays a key role in modulation of inflammation and the regulatory functions of immune cells, while also protecting cells against oxidative stress.

Institute of Biophysics of the Czech Academy of Sciences Kralovopolska 135 612 65 Brno Czech Republic

Institute of Biophysics of the Czech Academy of Sciences Kralovopolska 135 612 65 Brno Czech Republic Department of Animal Physiology and Immunology Institute of Experimental Biology Masaryk University Kotlarska 2 611 37 Brno Czech Republic

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