Excessive exposure of solar ultraviolet (UV) radiation, particularly its UV-B component, to humans causes many adverse effects that include erythema, hyperplasia, hyperpigmentation, immunosuppression, photoaging and skin cancer. In recent years, there is increasing use of botanical agents in skin care products. Pomegranate derived from the tree Punica granatum contains anthocyanins (such as delphinidin, cyanidin and pelargonidin) and hydrolyzable tannins (such as punicalin, pedunculagin, punicalagin, gallagic and ellagic acid esters of glucose) and possesses strong antioxidant and anti-inflammatory properties. Recently, we have shown that pomegranate fruit extract (PFE) possesses antitumor promoting effects in a mouse model of chemical carcinogenesis. To begin to establish the effect of PFE for humans in this study, we determined its effect on UV-B–induced adverse effects in normal human epidermal keratinocytes (NHEK). We first assessed the effect of PFE on UV-B–mediated phosphorylation of mitogen-activated protein kinases (MAPK) pathway in NHEK. Immunoblot analysis demonstrated that the treatment of NHEK with PFE (10–40 μg/mL) for 24 h before UV-B (40 mJ/cm2) exposure dose dependently inhibited UV-B–mediated phosphorylation of ERKl/2, JNK1/2 and p38 protein. We also observed that PFE (20 μg/mL) inhibited UV-B–mediated phosphorylation of MAPK in a time-dependent manner. Furthermore, in dose- and time-dependent studies, we evaluated the effect of PFE on UV-B–mediated activation of nuclear factor kappa B (NF-κB) pathway. Using Western blot analysis, we found that PFE treatment of NHEK resulted in a dose- and time-dependent inhibition of UV-B–mediated degradation and phosphorylation of IκBα and activation of IKKα. Using immunoblot analysis, enzyme-linked immunosorbent assay and electrophoretic mobility shift assay, we found that PFE treatment to NHEK resulted in a dose- and time-dependent inhibition of UV-B–mediated nuclear translocation and phosphorylation of NF-κB/p65 at Ser536. Taken together, our data shows that PFE protects against the adverse effects of UV-B radiation by inhibiting UV-B–induced modulations of NF-κB and MAPK pathways and provides a molecular basis for the photochemopreventive effects of PFE.
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Vol. 81 • No. 1