Antiinflammatory and antiangiogenic effect of a novel Fusion Protein "SLPI and Cementoin" targeting NF-{kappa}B pathway

SALICA, JP; CHULUYAN, E; GUERRIERI, D; ORTIZ, G; GALLO, JE

ORLANDO

Congreso; ARVO ANNUAL MEETING; 2014

ASSOCIATION FOR RESEARCH IN VISION AND OPHTHALMOLOGY

Abstract Purpose: We have previously demonstrated the anti-inflammatory and antiangiogentic effect of the fusion protein (SLPI and Cementoin) on a rat corneal alkali injury model. In this study we aimed at evaluating the effect of the fusion protein on the NF-{kappa}B (nuclear factor kappa-light-chain-enhancer of activated B cells) pathway during the acute inflammation process on a rat corneal alkali injury model. Methods: An alkali injury was produced with a filter paper of 3 mm with 1 N NaOH during 40 seconds in the right cornea of 27 Sprague-Dowley rats under general anesthesia. They were treated with FP-MC (0,2 ug/ul; n=9), SLPI (0,2 ug/ul; n=9) or vehicle (10ul; n=9) topically, four times a day. Three rats of each group were sacrificed at 6 hours, 4 days and 10 days after injury. Corneas were processed for Western Blot analysis and primary antibodies against the canonical and non canonical NF-kB pathway were used. Results: We found that the alkali injury-induced phosphorylation of nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha (I{kappa}Bα) and nuclear translocation of p65 were impaired by the fusion protein in comparison to the SLPI and Buffer treatment. Conclusions: The fusion protein keeps the property of its constituent serpin, showing an inhibition of the NF-kB pathway. Targeting NF-kB pathway is one of the proven mechanisms of action of this novel protein with anti-inflammatory and antiangiogentic effect.