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Home » Archive » 2020

TDK conference 2020

In vitro protective effect of luteolin against bacterial destruction on the intestinal epithelium
Móritz Alma Virág - year 6
University of Veterinary Medicine Budapest, Department of Pharmacology and Toxicology
Supervisors: Dr. Orsolya Farkas, Dr. Dóra Kovács

Abstract:

Nowadays, decreasing the development of antibiotic resistance is one of the most important challenges in both human- and veterinary medicine. Numerous EU guidelines were issued on prudent antibiotic usage, and an important goal of the 2019/6 EU regulation on veterinary medicinal products –to be entered in force in 2022- is to further regulate antibiotic usage of practicing veterinarians. Thereby many researchers aim to find alternative substances for substitution or supplementation of antibiotic therapy. We have conducted experiments with an antibacterial and antioxidant flavonoid compound, luteolin, which showed promising properties in human studies, therefore might serve as an antibiotic alternative in veterinary medicine.

In industrial livestock-raising, animal health is threatened by inflammation and oxidative stress in their intestinal epithelial cells caused by bacterial infections. Based on human research, we expected luteolin to be an effective antioxidant that could reduce harmful effects of bacteria. To prove this, amount of intracellular reactive oxygen species (ROS) were measured in vitro in porcine intestinal epithelial cell culture using DCFH-DA method, in a state of oxidative stress induced by bacterial endotoxin (LPS). IPEC-J2 cells were cultured on membrane inserts as well, as bacterial infections can damage barrier function of the intestine. FD4 fluorescent dye was used on these samples to determine whether luteolin could help in preserving the integrity under stress induced by LPS. Furthermore, bacteriostatic effect of luteolin was assessed by determining its minimal inhibitory concentration (MIC) against 8-8 Escherichia coli and Salmonella Typhimurium strains.

Our experiments showed that luteolin (at 25 µg/ml, 50 µg/ml and 100 µg/ml concentration) in vitro can reduce intercellular ROS in cells treated with S. Typhimurium LPS. In the aforementioned concentrations, it could also protect the integrity of intestinal epithelial cell layer against LPS treatment. Luteolin was able to inhibit bacterial growth at 256 µg/ml concentration.

In the future we are planning to continue examining antioxidant and integrity-protecting effects of luteolin on bacteria-intestinal epithelial cell co-cultures as well as conducting experiments with antibiotics to discover their possible synergistic effect with luteolin.



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