A bélhámsejtek oxidatív stressz okozta gyulladásának gátlása probiotikus tejsavbaktériumokkal: hatásmechanizmus és alkalmazhatóság antibiotikum-helyettesítő hozamfokozásra választási malacokon = Attenuation of oxidative stress-induced inflammation by probiotic lactobacilli in intestinal epithelial cells: mechanism of action and application as antibiotic-substitute in weanling pigs

Az akut oxidativ stressz es a probiotikumok hatasanak vizsgalatahoz kollagennel bevont polieszter membran inzerten tenyesztett, ujszulott sertes jejunum eredetű IPEC-J2 sejtvonalat alkalmaztunk. A peroxiddal kezelt IPEC-J2 sejtekben a gyulladasos citokinek (IL-8 es TNF-?) es a vedő hatasu hősokk feherje 70 (Hsp 70) relativ genexpressziojat kvantitativ real-time PCR-rel hataroztuk meg a probiotikus bakteriumtorzsek (Lactobacillus plantarum 2142, Lactobacillus casei Shirota, Bifidobacterium animalis subsp. lactis BB-12, Bacillus amyloliquefaciens CECT 5940, Enterococcus faecium CECT 4515) feluluszojanak (SMSz), illetve tejsav hozzaadasat kovetően. A Lactobacillus plantarum 2142 SMSz hatekonyan csokkentette az IL-8 es a TNF-? szintjet illetve novelte a Hsp70 genexpressziojat. Kimutattuk, hogy a gyulladascsokkentő hatas alapja nem a feluluszo es a hidrogen-peroxid kozotti kolcsonhatas, ezert a Lactobacillus plantarum 2142 gyulladascsokkentő hatasanak alapja nem az SMSz peroxid bontasa. Eredmenyeink szerint a Lactobacillus plantarum 2142 altal termelt tejsav (racem, L-es D-tejsav) nem csokkentette szignifikansan a gyulladasos citokinek expressziojat. Feherjeanalitikai, elvalasztastechnikai modszerek segitsegevel sikerult kimutatni a Lactobacillus plantarum 2142 altal termelt elterő molekulatomegű feherje jellegű anyagok jelenletet, amelyeknek szerepe lehet a gyulladascsokkentő hatas kifejteseben. | The experiments focused on acute oxidative stress and the effects of probiotics were performed on non-carcinogenic jejunal epithelial cell line of neonatal unsuckled piglet, IPEC-J2 cells cultured on collagen-coated polyester membrane inserts. In peroxide-treated IPEC-J2 cells relative gene expression levels of inflammatory cytokines (IL-8 es TNF-?) and protective heat shock protein 70 (Hsp 70) were determined by qRT-PCR in the presence and in the absence of SCSs from five bacterial strains (Lactobacillus plantarum 2142, Lactobacillus casei Shirota, Bifidobacterium animalis subsp. lactis Bb-12, Bacillus amyloliquefaciens CECT 5940, Enterococcus faecium CECT 4515) and lactic acid. Our data revealed that only the Lactobacillus plantarum 2142 had significant lowering effect on inflammatory cytokine levels (IL-8 es TNF-?) and it could increase the gene expression of Hsp 70. We proved that the no chemical reaction occurs between SCS and hydrogen-peroxide, which excludes the possibility that the antiinflammatory effect of Lactobacillus plantarum 2142 SCS could be attributed to its direct peroxide-decomposing activity. According to our results, lactic acid (racemic, D-and L-lactic acid) at different concentrations had not got any protective effect in quenching upregulation of proinflammatory cytokines. Peptide-like components present in SCS of Lactobacillus plantarum 2142, which are supposed to possess antiinflammatory effect, were detected by gel and capillary zone electrophoresis.