Abstract
The combination of ascorbic acid and rutin is frequently used in oral preparations. However, despite numerous protective effects of each component individually, their combined effect on ultraviolet (UV)-irradiated skin cells has never been evaluated. The aim of this study was to evaluate the combined effect of ascorbic acid and rutin on human keratinocytes and fibroblasts exposed to UVA and UVB radiation. Skin keratinocytes and fibroblasts exposed to UVA and UVB radiation were treated with ascorbic acid or/and rutin. The total antioxidant properties of both components, as well as their effect on cellular pro- and antioxidant status, lipid and protein oxidation, transmembrane transport, and pro-inflammatory and pro/antiapoptotic protein expression were measured. The combination of ascorbic acid and rutin had higher antioxidant properties compared to the activity of the single compound alone, and showed a stronger effect against UV-induced reactive oxygen species generation. The ascorbic acid and rutin combination also showed increased antioxidant enzyme activity (catalase, superoxide dismutase, thioredoxin reductase), which was impaired following UV irradiation. Moreover, ascorbic acid additional stimulated UV-induced bilitranslocase activity responsible for rutin transport, and therefore favored rutin effect on Nrf2 pathway, simultaneously differentiating the reaction of keratinocytes and fibroblasts. In keratinocytes, Nrf2 is strongly activated, while in fibroblasts decreased Nrf2 activity was observed. Used mixture, also significantly silenced UV-induced expression of pro-inflammatory factor NFκB and pro-apoptotic proteins such as caspases 3, 8, and 9. These results showed that ascorbic acid and rutin are complementary in their antioxidant actions, transport and signaling functions. Their combined antioxidant, antiinflammatory and antiapoptotic actions suggest rutin and ascorbic acid are a potentially cytoprotective team against UV-induced skin damage.
Highlights
Under physiological conditions, skin cells are characterized by the constant generation of reactive oxygen species (ROS), as well as the well-developed antioxidant system, which maintains the redox balance
Ascorbic A. and Rutin each increased the viability of UVA-irradiated keratinocytes by about 15%; the combination of Ascorbic A. + Rutin improved the viability of UVA- and UVB-irradiated keratinocytes by 20% and 35%, respectively
Both supplements alone and in combination did not have a significant effect on the viability of fibroblasts under standard conditions; Ascorbic A. + Rutin increased the viability of UVA- and UVB-irradiated fibroblasts by approximately 15% and 10%, respectively
Summary
Skin cells are characterized by the constant generation of reactive oxygen species (ROS), as well as the well-developed antioxidant system, which maintains the redox balance. Protein antioxidants act to protect skin cells by supporting Nrf, a ROS-dependent transcription factor, which initiates the transcription of cytoprotective and antioxidant genes [25]. Despite the maintenance of redox balance, frequent exposure of skin cells to the UVA and UVB radiation in natural sunlight stimulates pro-oxidative enzyme activity and impairs the action of antioxidants, resulting in oxidative stress [7, 21]. Previous studies have shown that exposure to UVA and UVB radiation leads to the activation of many factors involved in mitogen-activated protein (MAP)-dependent signaling kinases, including ERK1/2 and transcription factors dependent on redox potential, e.g., Nrf2 [24, 68]. The activation of multiple mechanisms associated with MAP kinases leads to the activation of the NFκB transcription factor and induces an inflammatory reaction [22, 54]
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