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TDK conference 2020Lewin Natalja - year 5 University of Veterinary Medicine Budapest, Department of Pharmacology and Toxicology Supervisors: Dr. Orsolya Farkas, Dr. Zita Karancsi Plant originated fermented wheat germ extract (FWGE) and proanthocyanidins, which can be found in high doses in grape seed extract (GSE), displays many beneficial properties, including a reduction of inflammation, strengthening of cell integrity or reducing oxidative stress. In this study, we analyzed protective effect of these substances on IPEC-J2 cell line, which is derived from jejunal epithelial cells of a neonatal piglet. These cells are not transformed, neither tumorous, therefore they can be used as a representative model of the physiological small intestine. The cells were cultured on semipermeable membrane inserts until they reached a confluent, differentiated monolayer. This was evaluated by measuring the transepithelial electrical resistance (TEER). After we reached appropriate TEER values, cells were treated with lipopolysaccharide (LPS), originated from Salmonella enterica serovar Typhimurium in a 10 µg/ml concentration to disrupt the cells’ barrier integrity. Simultaneously, we added FWGE (1% and 2% concentration) and GSE (50, 100 and 200 µg/ml concentration) to the cells to investigate their potential protective effect. For the control group, only pure cell culture medium was applied on the cells. To measure cell layer’s integrity, a fluorescent dye (fluorescein isothiocyanate-dextran – FD4) was added in the apical compartment of the cell culture wells and after 2, 4, and 24 hours, samples were taken from the basolateral compartments and fluorescent intensity was measured. LPS treatment could significantly (p<0.05) impair the cell’s barrier integrity compared to the control, but this effect was significantly (p<0.05) counteracted by both protective compounds (FWGE and GSE). Based on these results, we can assume that these natural plant extracts have beneficial effect not only on cell layer integrity in vitro, but on the intestinal epithelium in vivo as well. However, to prove this assumption, further in vivo investigations are necessary. List of lectures |