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Home » Archive » 2013

TDK conference 2013

A probiotic approach for the prevention of LPS-induced inflammation in the IPEC-J2 intestinal epithelial cell model
Piros Andrea Gabriella - year 4
SzIU, Faculty of Veterinary Science, Department of Pharmacology
Supervisors: Erzsébet Pásztiné Gere, Dr. Gálfi Péter

Abstract:

Advantages of probiotics’ application as growth-promoters have been proven in several cases. Underlying mechanisms behind beneficial effects of probiotics in the prevention and treatment of enteric infections have not been fully revealed. The aim of the study was the developement of in vitro test system which is suitable for tracing inflammatory processes triggered by the Gram negative cell wall component, lipopolysaccharide (LPS) treatment and investigation of potential anti-inflammatory effect of probiotics and their metabolites at cellular level.

To model complex interactions occuring in small intestine IPEC-J2 cell monolayer derived from newborn unsuckled piglet’s jejunum was cultured on polyester membrane insert. The effect of LPS treatment on the cell monolayer’s integrity was followed by measurement of transepithelial electrical resistance and assessment of apico-basolateral transport of fluorescein isothiocyanate labelled dextran (FD4). The formation of reactive oxygen species in LPS triggered inflammatory processes was detected by Amplex-red based fluorimetric measurements. During my experiments I studied the effect of Lactobacillus plantarum 2142 and Bacillus licheniformis CECT 4536 spent culture supernatants and sodium-n-butyrate how they can protect enterocytes from LPS-triggered cellular damage.

IPEC-J2 cell-system with distinct apical and basolateral compartments appeared to be reliable model to observe intestinal physiological and pathological conditions in vitro. The LPS administration without leading to significant ROS production could increase paracellular permeability through cell monolayer which was quantified by the elevated rate of FD4 apico-basolateral transport. Low dose Bacillus licheniformis CECT 4536 spent culture supernatant could decrease FD4 traffic by the strenghtening of intestinal monolayer integrity.

The aim of further studies is to examine intestinal barrier reinforcement ability and anti-inflammatory effect of several other probiotics and introduce their isolated metabolites in veterinary medicine.



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