KEY WORDS:Caspase activity  Glutathione balance In vitro digestion  Neurodegenerative diseases Wild blackberries

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L. Tavares  I. Figueira  H. L. A. Vieira 
P. M. Alves  R. B. Ferreira  C. N. Santos (&)
Instituto de Tecnologia Quı´mica e Biolo´gica, Universidade Nova
de Lisboa, Av. da Repu´blica, 2780-157 Oeiras, Portugal
G. J. McDougall  D. Stewart
Environmental and Biochemical Science Group, Enhancing
Crop Productivity and Utilisation Theme, The James Hutton
Institute, Dundee DD2 5DA, Scotland, UK
H. L. A. Vieira  P. M. Alves  C. N. Santos
Instituto de Biologia Experimental e Tecnolo´gica,
Apartado 12, 2781-901 Oeiras, Portugal
H. L. A. Vieira
CEDOC@IGC, Faculdade de Cieˆncias Me´dicas,
UNL, 1169-056 Lisbon, Portugal
D. Stewart
School of Life Sciences, Heriot Watt University,
Edinburgh EH14 4AS, Scotland, UK
R. B. Ferreira
Departamento de Botaˆnica e Engenharia Biolo´gica, Instituto
Superior de Agronomia, Universidade Te´cnica de Lisboa,
Tapada da Ajuda, 1349-017 Lisbon, Portugal

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ABSTRACT：Purpose Blackberry ingestion has been demonstrated to attenuate brain degenerative processes with the benefits ascribed to the (poly)phenolic components. The aim of this work was to evaluate the neuroprotective potential of two wild blackberry species in a neurodegeneration cell model and compare them with a commercial variety. Methods This work encompasses chemical characterization before and after an in vitro digestion and the assessment of neuroprotection by digested metabolites. Some studies targeting redox/cell death systems were also performed to assess possible neuroprotective molecular mechanisms. Results The three blackberry extracts presented some quantitative differences in polyphenol composition that could be responsible for the different responses in the neurodegeneration cell model. Commercial blackberry extracts were ineffective but both wild blackberries, Rubus brigantinus and Rubus vagabundus, presented neuroprotective effects. It was verified that a diminishment of intracellular ROS levels, modulation of glutathione levels and activation of caspases occurred during treatment. The last effect suggests a preconditioning effect since caspase activation was not accompanied by diminution in cell death and loss of functionality. Conclusions This is the first time that metabolites obtained from an in vitro digested food matrix, and tested at levels approaching the concentrations found in human plasma, have been described as inducing an adaptative response.