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Thymol and Thymus vulgaris extract protects human keratinocyte cell line (HaCaT) from UVA and UVB damage

Martina Mapelli, Rossella Calo, Laura Marabini.

Abstract
Objective: The aims of our study were to characterize ultraviolet (UV)A- and UVB-induced damages in a keratinocytes cell line (HaCaT), and to evaluate the protective capacities two plant-derivative compounds, namely Thymus vulgaris L leaf extract and thymol, its major component. A polyphenol rich diet has gained wide attention and it is now considered to be a protective agent for human skin, which can be over-exposed to environmental factors and in particular UV light.
Methods: Cells were pretreated for 1 h, in serum-free medium, with thymol (1 μg/ml) or Thymus vulgaris L (1.82 μg/ml) then exposed to different UVA (8-24 J/cm2) or UVB doses (0.016-0.72 J/cm2). Immediately after the UV exposure the intracellular redox status was evaluated by reactive oxygen species quantification and apoptotic events. Genotoxic aspects were evaluated 24 h after the end of irradiations using the alkaline comet assay and the immunostaining of phosphorylated H2AX histone protein (detected 1 h after the end of UV exposure).
Results: The pre-treatment of our experimental model with the two substances confirmed an antioxidant action and anti-apoptotic effect by reducing the cells percentage (sub-G1 phase). Furthermore, thymol and extract of Thymus vulgaris L were able to reduce genotoxic damage. The alkaline comet assay showed that the two substances were capable to decrease DNA damage. Also in this case, Thymus vulgaris L extract is more effective than thymol in decreasing genotoxicity markers.
Conclusions: Our results confirmed the more oxidant UVA and more genotoxic UVB effects. Regarding the protective effect of thymol and Thymus vulgaris L extract, data obtained proved their antioxidant and free-radical scavenging ability as known for phenolic (which our compounds belong to) and polyphenolic compounds. Thymol and mainly Thymus vulgaris L extract were also able to reduce the direct genotoxic damage.

Key words: Genotoxicity; phenolic compounds; ultraviolet radiations; thmol; Thymus vulgaris



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