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ArchiveKákonyi Ákos Richárd - year 3 University of Veterinary Medicine Budapest, Department of Physiology and Biochemistry Supervisors: Dr. Patrik Tráj, Dr. Gábor Mátis Nowadays bacterial resistance to antibiotics is a growing and worldwide threat. The irresponsible use of antibiotics has led to the emergence of resistant pathogens in both human and veterinary medicine, which we will no longer be able to defend against in the future. The use of feed supplements of natural origin may be an alternative to diminish the damage caused by certain bacteria in veterinary medicine. These can be used both as a treatment and as growth promotion, reducing the damage caused by infectious diseases. Since bacterial infection in most cases triggers an inflammatory response in the body, our research aimed to study the development of inflammatory processes caused by bacteria and the possibilities of influencing them. Our work aimed to investigate the pro-inflammatory effect of flagellin released from the bacterial flagellum, the organelle of motile bacteria. Furthermore, we intended to attenuate the induced inflammation with luteolin on chicken primary hepatocyte and non-parenchymal co-culture. The literature suggests a substantial role for flagellum and flagellin in the pathogenesis of Salmonella-induced diseases and the induction of inflammation. Luteolin is a plant-derived flavonoid widely found in herbs and fodder plants. There are recent studies on its antioxidant, antimicrobial, antitumor and anti-inflammatory activity in mammal species and certain cell cultures. Our cells were cultured in a medium supplemented with 250 ng/ml Salmonella Typhimurium-derived flagellin and 4 or 16 µg/ml luteolin for 24 hours. Cell metabolic activity, lactate dehydrogenase (LDH) activity, interleukin-6 (IL-6), interleukin-8 (IL-8) and hydrogen peroxide (H2O2) concentrations were then measured from the medium. Malondialdehyde (MDA) concentrations were determined from cell lysates. Flagellin induced a significant up-regulation of the concentration of the pro-inflammatory cytokine IL-8 in the culture medium, and thus flagellin-treated cell cultures may be a suitable model for studying the inflammatory process in vitro. Luteolin treatment at 4 µg/ml did not prove to be cell damaging as measured by metabolic activity and extracellular LDH activity, and significantly reduced the flagellin-induced increase of IL-8 concentration in our cultures. In addition, it had a reducing effect on the levels of medium H2O2 and cell lysate MDA. These results suggest that luteolin at lower concentrations may protect cells from an excessive inflammatory response and may act as an antioxidant to attenuate reactive oxygen species production and the resulting lipid peroxidation. List of lectures |