Disclosing the mechanisms underlying berries health promotion
CLOSED
Disclosing the mechanisms underlying berries health promotion in a neurodegeneration cell model
Background
Recent consumer's interest in berries is due to their highly advertised health impact. Evidences include protection in neurological diseases, in particular during aging.
The present proposal envisages disclosing molecular mechanisms underlying the beneficial health effect promoted by berries. To achieve this goal, new approaches will be addressed: i) a neurodegeneration cell model will be treated with physiological concentrations of berries digested metabolites [1]; ii) quasi-isogenic fruits will be used to allow the association of different mechanism to specific phytochemicals classes; and iii) cell mechanisms associated with survival/death, metabolism and redox status will be monitored. These new approaches come in line of thought that antioxidant phytochemicals are greatly altered during gastrointestinal digestion and the concentrations that reach serum and are bioavailable to brain are reduced comparing to the amounts ingested [2]. Accordingly to this, hardly phytochemicals could act in the brain as antioxidants per se, they are more prone to act trough modulation of signalling pathways and specific biological targets.
Genotype has being indicated as a major factor determining the chemical diversity of bioactive compounds content in the berries [3]. Cultivars of the same type of fruit have a very close constitution, being enriched or depleted in some phytochemicals classes. By this way, cultivars constitute quasi-isogenic foods. Using cultivars it will be associated different protective mechanism to specific berries phytochemicals classes.
Working plan
Task1- Neuroprotective potential of berries digested metabolites in a human cell line
Berry digested metabolites at serum relevant concentrations will be tested in a neurodegeneration cell model, already implemented in host lab [1]. The neuroprotection will be accessed by flow cytometry, monitoring plasmatic membrane integrity and mitochondrial transmembrane potential [1].
Task 2- Molecular mechanism underlying the neuroprotective effects This project will focus on three cellular processes that can be involved in prevention of neurodegeneration:
i. DEATH/SURVIVAL MECHANISMS will be assessed through flow cytometry/ fluorescent microscopy using PI and Annexin V conjugates as probes for NECROSIS and APOPTOSIS, respective.
ii. CELL METABOLISM will be monitorized for several metabolites traced by enzymatic assays, such as GLUCOSE, GLUTAMINE, LACTATE and ATP [4].
iii. REDOX METABOLIC alterations will be evaluated by glutathione balance (GSH/GSSG) assessed by HPLC with fluorimetric detection[5]. Compounds related to the glutathione metabolism will be used to reveal the importance of this antioxidant system and redox cell response.
References
[1] Tavares, L.; Figueira, I.; Macedo, D.; McDougall, G. J.; Leitão, M. C.; Vieira, H. L. A.; Stewart, D.; Alves, P. M.; Ferreira, R. B.; Santos, C. N. Food Chemistry, 2012, 131, 1443-1452.
[2] Manach, C.; Scalbert, A.; Morand, C.; Remesy, C.; Jimenez, L. Am J Clin Nutr, 2004, 79, 727-47.
[3] Anttonen, M. J.; Karjalainen, R. O. Journal of Food Composition and Analysis, 2005, 18, 759-769.
[4] Almeida, A. S.; Queiroga, C. S. F.; Sousa, M. F. Q.; Alves, P. M.; Vieira, H. L. A. Journal of Biological Chemistry, 2012, 287, 10761-10770.
[5] Kand'ár, R.; Záková, P.; Lotková, H.; Kucera, O.; Cervinková, Z. Journal of Pharmaceutical and Biomedical Analysis, 2007, 43, 1382-1387.
The present project is aligned with the aim of European project FP7-KBBE-2010-4 (265942) EUBerry.
Supervisors
Cláudia Nunes dos Santos (csantos@itqb.unl.pt)
Lucélia Rodrigues Tavares (ltavares@itqb.unl.pt)
Laboratory/Institution
