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TDK conference 2021

Antioxidant and anti-inflammatory effects of proanthocyanidins in porcine intestinal epithelial cell – bacterium co-culture
Gál Bella - year 5
University of Veterinary Medicine Budapest, Department of Pharmacology and Toxicology
Supervisor: Dr. Dóra Kovács


Nowadays, as a result of the abundant and inappropriate use of antibiotics, resistance in bacteria against these agents poses a potentially great risk in both veterinary and human medicine. The increasing prevalence of resistant strains can lead to treatment failure, which is especially important in case of infections caused by zoonotic bacteria such as Escherichia coli and Salmonella enterica ser. Typhimurium. This phenomenon can only be overcome with the ‘One Health’ concept, where the veterinary and human fields work together in finding solutions to decrease and optimise the usage of antibiotics and to find potential antibiotic alternatives that can help in the prevention and treatment of bacterial infections. Among many natural substances, flavonoids, including proanthocyanidins (PACs), might be used for this purpose due to their antioxidant, anti-inflammatory and antibacterial properties.

In this study, our aim was to test protective effects of grape seeds PACs in an in vitro model of gastrointestinal infections, which consisted of bacteria (E. coli and S. Typhimurium) of swine enteral origin and porcine intestinal epithelial cells (IPEC-J2 cell line). To establish this co-culture, initially, Neutral Red assay was used to determine the highest tolerable bacterial concentration that could be applied on IPEC-J2 cells without causing significant viability decrease. Afterwards, cells were treated with either E. coli or S. Typhimurium alone or in combination with PACs (50 or 100 µg/ml), and the amount of intracellular reactive oxygen species (IC ROS) were measured with 2’,7’ dichloro-dihydro-fluorescein diacetate (DCFH-DA) reagent, while interleukin-6 (IL-6) and interleukin-8 (IL-8) levels were determined with enzyme-linked immunosorbent assay (ELISA).

In our experiment, 1 hour treatment with 10⁶ CFU/ml E. coli and S. Typhimurium did not affect the viability of IPEC-J2 cells, therefore bacteria were used in this concentration during the further steps. PACs in 50 and 100 µg/ml concentrations exhibited the ability to reduce IC ROS level of IPEC-J2 cells when they were applied in combination with bacteria. Furthermore, S. Typhimurium caused an increase in IL-8 and IL-6 production of cells, which could also be alleviated with the application of PACs.

Our results showed that PACs from grape seeds were able to reduce the amount of oxidative stress and inflammatory markers in porcine intestinal epithelial cells treated with bacteria. Based on these observations, PACs might be eligible for the prevention and treatment of intestinal bacterial infections in swine, but further in vitro and in vivo studies are necessary to support their usage.

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