Ouabain is a steroid hormone that binds to the enzyme Na+, K+ - ATPase and stimulates different intracellular pathways controlling growth, proliferation and cell survival. IL-1 beta and TNF-alpha are pleiotropic molecules, conventionally regarded as pro-inflammatory cytokines with well-known effects in the immune system. In addition, IL-1 beta and TNF-alpha also play important roles in the nervous system including neuroprotective effects. Previous data from our group showed that ouabain treatment is able to induce an increase in retinal ganglion cell survival kept in mixed retinal cell cultures. The aim of this work was to investigate if IL-1 beta and TNF-alpha could be mediating the trophic effect of ouabain on retinal ganglion cells. Our results show that the trophic effect of ouabain on retinal ganglion cell was inhibited by either anti-IL-1 beta or anti-TNF-alpha antibodies. In agreement, IL-1 beta or TNF-alpha increased the retinal ganglion cells survival in a dose-dependent manner. Accordingly, ouabain treatment induces a temporal release of TNF-alpha and IL-1 beta from retinal cell cultures. Interestingly, TNF-alpha and IL-1 beta regulate each other intracellular levels. Our results suggest that ouabain treatment triggers the activation of TNF-alpha and IL-1 beta signaling pathways leading to an increase in retinal ganglion cell survival. (C) 2016 Published by Elsevier Inc.