No abstract

Reactive oxygen species are implicated in cell and tissue damage in a number of diseases including acute and chronic inflammation of the gut. Effects of H(2)O(2) exposure on non-carcinogenic porcine epithelial cell line, IPEC-J2 cells cultured on collagen-coated membrane inserts were monitored based on transepithelial electrical resistance (TER) change, extent of necrotic cell damage, gene expression of inflammatory cytokines IL-8 and TNF-α. Furthermore, the junction proteins claudin-1 and E-cadherin were also investigated by immunohistochemistry. Peroxide (1mM) increased IL-8 and TNF-α gene expression levels significantly allowing 1 h recovery time without affecting the cellular distribution of junction proteins, TER and cell survival rate. In conclusion, the IPEC-J2 cell line on membrane insert was introduced as a fast and reliable investigation tool for oxidative stimuli-triggered intestinal inflammation and in the future as a screening method for antioxidant and probiotic candidates.

Recently, there has been a growing interest to replace antibiotics' administration with the application of probiotics. The aim of our investigations was to reveal the influence of spent culture supernatant of Lactobacillus plantarum 2142 on the response of enterocytes to oxidative stress, and the spent culture supernatant's ability to protect them from oxidative injury. The experiments were performed on non-carcinogenic porcine epithelial cell line, IPEC-J2 isolated from a neonatal piglet and on human colon adenocarcinoma cell line, Caco-2. The cells cultured on membrane inserts were treated with millimolar hydrogen peroxide solution to provoke oxidative stress. The peroxide-triggered cell response profile was evaluated via determination of change in transepithelial electrical resistance, quantification of extent of cell death by 4',6-diamidino-2 phenylindole (DAPI) staining and via estimation of proinflammatory cytokine, IL-8 production using ELISA technique. Non-starter lactobacilli supernatant-mediated inhibition of peroxide-triggered upregulation of IL-8 production confirmed the antiinflammatory properties of active metabolites produced by Lactobacillus plantarum 2142 in acute oxidative stress.