UVA radiation is one of the main harmful factor that penetrates all human skin layers and influencing communication between the cells that build them. To prevent UVA-induced damage, there is a constant search for compounds that could protect all skin cells, as well as restoring homeostasis in their communication. Therefore, the aim of this study was to evaluate the effect of 3-O-ethyl ascorbic acid (EAA) on the intracellular proteome of co-cultured keratinocytes and fibroblasts after UVA irradiation and on the profile of proteins released into the medium by both cell types. Proteomic approach allowed to identify significantly modified by UVA as well as EAA proteins. In keratinocytes UVA radiation enhanced expression of pro-inflammatory and pro-proliferative/keratinizing proteins and decreased anti-apoptotic and antioxidant proteins, while in fibroblasts UVA radiation induced expression mainly of proinflammatory proteins, simultaneously decreasing the level of proteins involved in antioxidant response, as well as growing factors. Similar effect was observed in the medium of co-cultured cells, where increase in pro-inflammatory protein and decrease in growing factors were observed. Medium supplementation with EAA restored the levels of these proteins compared to control cultures. The results of this study show that EAA may act as a protector of epidermal and dermal cells, because it reduces the levels of pro-inflammatory proteins and increases the activity of the antioxidant system of skin cells (keratinocytes and fibroblasts), as well as normalizes intercellular signaling.