In vitro Anti-inflammatory and ProtectiveEffects of ibid???? on Intestinal Epithelial Cells

摘要

Aim: To define the putative anti-inflammatory and cytotoxic effects of ibidì?, a new phytotherapeutic formulation composed of three extracts: Punica granatum L, pericarpum; Boswellia serrata Roxb., resina; Curcuma longa L,. rhizome, using Caco-2 cells, an in vitro model of human intestinal epithelium. Methodology: cytotoxicity and capacity of ibidì? to induce cell proliferation were assessed respectively by 2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2H-Tetrazolium-5-Carboxanilide (XTT) assay and nucleotide 5-bromo-2’-deoxyuridine (BrdU) incorporation. Cell migration was evaluated by scratch wound assay. COX-2, IL-6, IL-8 and MCP-1 protein levels were measured in the supernatant of cells stimulated with or without TNF-alfa or IL-1 beta in presence or in absence of ibidì? using ELISA assays. Finally, the influence of ibidì? on the integrity, paracellular permeability, and viability of Caco-2 cell monolayers was monitored by measuring the transepithelial electrical resistance (TEER) in presence or in absence of TNF-??? stimulation.Results: No dose-response toxicity was observed after 48 h incubation with ibidì?. Interestingly the cell proliferation rate was generally lower in presence of ibidì? than vehicle at all concentrations tested, while ibidì? had no effects on cell migration. Ibidì? markedly inhibited TNF-alfa-induced production of IL-8 at all concentrations tested in a dose-response manner, while that of IL-6 and MCP-1 only at highest ibidì? concentrations. Importantly ibidì? in a range of concentration between 145 and 9 μg/ml not only abrogated TNF-alfa-dependent TEER depression, but also promoted higher resistance values than untreated cells.Conclusion: These data demonstrate that ibidì? exerts anti-inflammatory and protective effects on intestinal epithelial cells by blocking the production of IL-8, IL-6 and MCP-1, and unveil that the synergism of the three extracts regulates epithelial barrier function.