title: Bioactivity of Fragaria vesca leaves through inflammation, proteasome and autophagy modulation.
abstract: BACKGROUND
  Fragaria vesca leaves have been used in folk medicine for the treatment of several diseases, namely gastrointestinal, cardiovascular and urinary disorders, which could be related with the potential anti-inflammatory properties of the extract. This work aims to disclose the bioactivity and the underlying action mechanism of an extract from Fragaria vesca leaves in order to support its traditional uses.
  
  
  METHODS
  A hydroalcoholic extract was prepared from Fragaria vesca leaves and its anti-inflammatory potential was evaluated through inhibition of nitric oxide production and expression of several pro-inflammatory proteins in lipopolysaccharide-triggered macrophages. Nitric oxide scavenger activity was also assessed using a standard nitric oxide donor. Since numerous inflammatory proteins are tightly regulated by ubiquitination and proteasomal degradation, the putative effect of the extract on these cellular proteolytic pathways was also disclosed. The phytochemical characterization was performed by HPLC-PDA-ESI/MSn and compared with an infusion prepared according to the traditional method.
  
  
  RESULTS
  For non-cytotoxic concentrations (80 and 160µg/mL) the extract inhibited nitrite production, probably due to a direct nitric oxide scavenging. Furthermore, inhibition of proteasome activity was verified, leading to accumulation of ubiquitinated proteins. The extract also increased the conversion of the microtubule-associated protein light chain LC3-I to LC3-II, a marker of autophagy. Polyphenols, namely ellagitannins, proanthocyanidins, and quercetin and kaempferol glucuronide derivatives were identified in Fragaria vesca leaves extract. Most of the identified phenolic compounds matched with those found in traditional preparation, the infusion.
  
  
  CONCLUSIONS
  The extract has a direct nitric oxide scavenging activity giving support to the traditional use of this plant for the treatment of inflammatory disorders. Furthermore, the extract affects the proteolytic systems but its role in cancer treatment requires further studies.
authors.affiliation: Center for Pharmaceutical Studies, Faculty of Pharmacy, University of Coimbra, Pólo das Ciências da Saúde, Azinhaga de Santa Comba, 3000-548 Coimbra, Portugal; Center for Neurosciences and Cell Biology, University of Coimbra, Largo Marquês de Pombal, 3004-517 Coimbra, Portugal; Faculty of Pharmacy, University of Coimbra, Pólo das Ciências da Saúde, Azinhaga de Santa Comba, 3000-548 Coimbra, Portugal.
authors.forename: Joana
authors.initials: J
authors.lastname: Liberal
authors.affiliation: Center for Pharmaceutical Studies, Faculty of Pharmacy, University of Coimbra, Pólo das Ciências da Saúde, Azinhaga de Santa Comba, 3000-548 Coimbra, Portugal; Center for Neurosciences and Cell Biology, University of Coimbra, Largo Marquês de Pombal, 3004-517 Coimbra, Portugal; Faculty of Pharmacy, University of Coimbra, Pólo das Ciências da Saúde, Azinhaga de Santa Comba, 3000-548 Coimbra, Portugal.
authors.forename: Vera
authors.initials: V
authors.lastname: Francisco
authors.affiliation: Center for Pharmaceutical Studies, Faculty of Pharmacy, University of Coimbra, Pólo das Ciências da Saúde, Azinhaga de Santa Comba, 3000-548 Coimbra, Portugal; Center for Neurosciences and Cell Biology, University of Coimbra, Largo Marquês de Pombal, 3004-517 Coimbra, Portugal; Faculty of Pharmacy, University of Coimbra, Pólo das Ciências da Saúde, Azinhaga de Santa Comba, 3000-548 Coimbra, Portugal.
authors.forename: Gustavo
authors.initials: G
authors.lastname: Costa
authors.affiliation: Center for Pharmaceutical Studies, Faculty of Pharmacy, University of Coimbra, Pólo das Ciências da Saúde, Azinhaga de Santa Comba, 3000-548 Coimbra, Portugal; Department of Environment, Polytechnic Institute of Viseu, Campus Politécnico de Repeses, 3504-510 Viseu, Portugal.
authors.forename: Artur
authors.initials: A
authors.lastname: Figueirinha
authors.affiliation: Center for Pharmaceutical Studies, Faculty of Pharmacy, University of Coimbra, Pólo das Ciências da Saúde, Azinhaga de Santa Comba, 3000-548 Coimbra, Portugal.
authors.forename: Maria Teresa
authors.initials: MT
authors.lastname: Amaral
authors.affiliation: Centre of Ophthalmology and Vision Sciences, Institute for Biomedical Imaging and Life Sciences, IBILI, Faculty of Medicine, University of Coimbra, 3000-548 Coimbra, Portugal.
authors.forename: Carla
authors.initials: C
authors.lastname: Marques
authors.affiliation: Centre of Ophthalmology and Vision Sciences, Institute for Biomedical Imaging and Life Sciences, IBILI, Faculty of Medicine, University of Coimbra, 3000-548 Coimbra, Portugal.
authors.forename: Henrique
authors.initials: H
authors.lastname: Girão
authors.affiliation: Center for Neurosciences and Cell Biology, University of Coimbra, Largo Marquês de Pombal, 3004-517 Coimbra, Portugal; Faculty of Pharmacy, University of Coimbra, Pólo das Ciências da Saúde, Azinhaga de Santa Comba, 3000-548 Coimbra, Portugal.
authors.forename: Maria Celeste
authors.initials: MC
authors.lastname: Lopes
authors.affiliation: Center for Neurosciences and Cell Biology, University of Coimbra, Largo Marquês de Pombal, 3004-517 Coimbra, Portugal; Faculty of Pharmacy, University of Coimbra, Pólo das Ciências da Saúde, Azinhaga de Santa Comba, 3000-548 Coimbra, Portugal.
authors.forename: Maria Teresa
authors.initials: MT
authors.lastname: Cruz
authors.affiliation: Center for Pharmaceutical Studies, Faculty of Pharmacy, University of Coimbra, Pólo das Ciências da Saúde, Azinhaga de Santa Comba, 3000-548 Coimbra, Portugal; Center for Neurosciences and Cell Biology, University of Coimbra, Largo Marquês de Pombal, 3004-517 Coimbra, Portugal; Faculty of Pharmacy, University of Coimbra, Pólo das Ciências da Saúde, Azinhaga de Santa Comba, 3000-548 Coimbra, Portugal. Electronic address: mtpmb@ff.uc.pt.
authors.forename: Maria Teresa
authors.initials: MT
authors.lastname: Batista
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000008:administration & dosage / Q000302:isolation & purification / Q000494:pharmacology*; D001343:Autophagy / Q000187:drug effects; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004305:Dose-Response Relationship, Drug; D031985:Fragaria / Q000737:chemistry*; D016166:Free Radical Scavengers / Q000008:administration & dosage / Q000302:isolation & purification / Q000494:pharmacology; D007249:Inflammation / Q000188:drug therapy*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism; D008519:Medicine, Traditional; D051379:Mice; D009569:Nitric Oxide / Q000378:metabolism; D010936:Plant Extracts / Q000008:administration & dosage / Q000494:pharmacology*; D018515:Plant Leaves; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism
chemical_list: D000893:Anti-Inflammatory Agents; D016166:Free Radical Scavengers; D008070:Lipopolysaccharides; D010936:Plant Extracts; D009569:Nitric Oxide; D046988:Proteasome Endopeptidase Complex
keywords: Autophagy; Fragaria vesca leaves; Inflammation; Nitric oxide scavenger; Polyphenols; Ubiquitin-Proteasome system