title: Vitis vinifera seeds extract for the modulation of cytosolic factors BAX-α and NF-kB involved in UVB-induced oxidative stress and apoptosis of human skin cells.
abstract: OBJECTIVE
  The depletion of the ozone layer allows overexposure of the skin to UV radiation, which is prolonged due to the increasing life expectancy, together with inappropriate life habits contribute to the increasing incidence of cutaneous malignancies. Plant extracts with antioxidant capacities are frequently employed as a means to protect skin against ultraviolet (UV) radiations, thus preventing skin cancers. In the present study we assessed a red grape seed extract (GSE) potential capacities to reduce ultraviolet B (UVB) radiation-induced reactive oxygen species (ROS) and subsequent apoptosis in a human keratinocytes cell line (HaCaT). We identified molecules and pathways modulated by the GSE through which this may exert its photoprotective effect.
  
  
  METHODS
  The GSE was standardized according to its polyphenolic content and the most important biologically active compounds, such as epigallocatechin and epicatechin, catechin hydrate, procyanidin B and gallic acid were evidenced by high-performance liquid chromatography. According to the plant extract cytotoxicity on the HaCaT cell line, two concentrations were selected for testing from the non-toxic range: GSE1 (37.5 μgEqGA/ml) and GSE2 (75 μgEqGA/ml). The level of ROS was evaluated with CM-H2DCFDA assay, while apoptosis, Bax-α and NF-kβ p65 proteins with ELISA and confirmed by western-blot.
  
  
  RESULTS
  Both concentrations of the extract decreased the level of ROS in UVB-irradiated keratinocytes (p<0.001), whereas apoptosis and Bax-α pro-apoptotic protein were only reduced by the higher concentration (GSE2). The NF-kB p65 protein level registered increasing values in time after UVB exposure of the cells, while the tested plant extract re-established its level when its smaller concentration was used (GSE1).
  
  
  CONCLUSIONS
  These results encourage further studies on this extract in order to identify other molecules and pathways through which this extract might exert its beneficial effects and also recommend its use as a potential photoprotective agent.
authors.affiliation: Department of Physiology/Functional Sciences, Iuliu Hatieganu University of Medicine and Pharmacy, Cluj-Napoca, Romania; Emergency Military Hospital, Cluj-Napoca, Romania.
authors.forename: Hana
authors.initials: H
authors.lastname: Decean
authors.affiliation: Prof. Dr. I. Chiricuta Oncology Institute, Cluj-Napoca, Romania.
authors.forename: Eva
authors.initials: E
authors.lastname: Fischer-Fodor
authors.affiliation: Prof. Dr. I. Chiricuta Oncology Institute, Cluj-Napoca, Romania.
authors.forename: Corina
authors.initials: C
authors.lastname: Tatomir
authors.affiliation: Prof. Dr. I. Chiricuta Oncology Institute, Cluj-Napoca, Romania.
authors.forename: Maria
authors.initials: M
authors.lastname: Perde-Schrepler
authors.affiliation: Babes-Bolyai University, Cluj-Napoca, Romania.
authors.forename: Lidia
authors.initials: L
authors.lastname: Somfelean
authors.affiliation: Prof. Dr. I. Chiricuta Oncology Institute, Cluj-Napoca, Romania; Iuliu Hatieganu University of Medicine and Pharmacy, Cluj-Napoca, Romania.
authors.forename: Claudia
authors.initials: C
authors.lastname: Burz
authors.affiliation: Iuliu Hatieganu University of Medicine and Pharmacy, Cluj-Napoca, Romania.
authors.forename: Tudor
authors.initials: T
authors.lastname: Hodor
authors.affiliation: Iuliu Hatieganu University of Medicine and Pharmacy, Cluj-Napoca, Romania; Dermatology Clinical Hospital, Cluj-Napoca, Romania.
authors.forename: Remus
authors.initials: R
authors.lastname: Orasan
authors.affiliation: Prof. Dr. I. Chiricuta Oncology Institute, Cluj-Napoca, Romania.
authors.forename: Piroska
authors.initials: P
authors.lastname: Virag
keywords: Bax-α; NF-kB; ROS; apoptosis; red grape seed extract