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Study of the Health Promoting Properties of the Tropical Highland

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Study of the Health Promoting Properties of the Tropical Highland Study of the health promoting properties of the tropical highland blackberry (Rubus adenotrichos) and the impact of digestion and pasteurization processes Gabriela Azofeifa To cite this version: Gabriela Azofeifa. Study of the health promoting properties of the tropical highland blackberry (Rubus adenotrichos) and the impact of digestion and pasteurization processes. Agricultural sciences. Uni- versité Montpellier II - Sciences et Techniques du Languedoc, 2013. English. NNT : 2013MON20064. tel-01002531 HAL Id: tel-01002531 https://tel.archives-ouvertes.fr/tel-01002531 Submitted on 6 Jun 2014 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Abstract This study evaluated biological activities of blackberries (Rubus adenotrichos Schltdl.) and the effects of digestion or pasteurization processes on these activities. The blackberries phenolic extract showed an ORAC value of 4.34 ± 0.14 mmol TE/g, higher than those of quercetin and ellagic acid. It also protected liposomes and liver homogenates against lipid peroxidation; with IC50 of 7.0 ± 0.5 and 20.3 ± 4.2 μg/mL, respectively. The blackberry polyphenols inhibited nitrite production in J774A.1 cells stimulated with LPS+IFNγ due to down-regulation of iNOS protein expression, suggesting an anti- inflammatory potential. The biological activities of the fruits could be affected by the industrial processes. The pasteurization of a blackberry juice caused a 7% decrease in the anthocyanins concentration of the highest temperature tested, 92 °C. For the same sample, a significant reduction of 27% and 15% were showed in the DPPH and NO scavenging capacity, respectively. However, no significant differences were evident neither for the peroxidation inhibitory capacity nor for the intracellular antioxidant activity after the pasteurization step. The antioxidant capacity is largely dependent of their bioavailability and because of this a blackberry juice was digested in vitro to obtain samples that represent the major stages of the digestion process. The gastric digestion had slight impact on polyphenolic composition while the pancreatin/bile digestion had marked effects on polyphenolic composition, particularly in the ratio anthocyanins-ellagitannins. These changes in phenolic composition caused in the dialyzed fraction a reduction of 33% and 35% in the activity against nitrogen derived radicals DPPH and NO respectively. As well the capacity to inhibit intracellular ROS was decrease by an 85% in the dialyzed fraction. Nevertheless, no significant differences were evident in the capacity to inhibit lipid peroxidation. Finally, this study used an in vivo model with diabetic rats to evaluate the effect of the consumption of a blackberry beverage. The blackberry intake provoked a decrease in plasmatic glucose (-48.6%), triglycerides (-43.5%) and cholesterol (-28.6%) levels in the diabetic rats with respect to the diabetic controls animals. The diabetic rats which consumed blackberry, improved (+7%) the plasma antioxidant capacity (PAC) values and reduced the levels of lipid peroxidation in plasma (-19%) and in kidney tissue (-23%). This study suggests that blackberry could be considered as a candidate for the group of functional foods even when considering the bioavailability aspects of the phenolic compounds and the pasteurization effects. Key words: Blackberry, Rubus adenotrichos, antioxidant activity, anti-inflammatory activity, diabetes mellitus, pasteurization, bioavailability. i Résumé Dans cette étude nous avons évalué les activités biologiques de la mûre tropicale de montagne (Rubus adenotrichos Schltdl.) et les effets de la digestion ou des procédés de pasteurisation sur ces activités. L’extrait polyphénolique de mûre présente un pouvoir antioxydant déterminé par la méthode ORAC de 4.34 ± 0.14 mmol TE/g, plus élevé que celui de la quercétine et de l'acide ellagique. L’extrait de mûre présente des propriétés protectrices des liposomes et des homogénats de foie contre la peroxydation lipidique avec des IC50 de 7.0 ± 0.5 et 20.3 ± 4.2 mg/ml, respectivement. Les polyphénols de mûre inhibent la production de nitrite dans par les macrophages murins J774A.1 stimulées par le mélange LPS, IFN. Cette activité est confirmée par la diminution du taux de NO synthase inductible dans ces mêmes cellules. Les activités biologiques des fruits pourraient être affectés par les procédés industriels. Le processus de pasteurisation du jus de mûre a provoqué une diminution d’environ 7% de la concentration en anthocyanes pour la température la plus élevée évalué, 92°C. Pour le même échantillon, une réduction significative de 27% de l’activité anti-oxydante mesurée par la méthode DPPH et de 16% de la capacité de complexassions des radicaux NO. Toutefois, aucune différence significative n’a été mise en évidence autant pour la capacité d'inhibition de la peroxydation que pour l'activité anti-oxydante intracellulaire. Les effets in vivo sont largement dépendants de la biodisponibilité des composés phénoliques et de ce fait un jus de mûre a été digéré in vitro pour obtenir des échantillons qui représentent les grandes étapes du processus de digestion. La digestion gastrique a peu d’influence au contraire de la digestion intestinale sur la composition polyphénolique de l’extrait de mûre en particulier au niveau du rapport anthocyanines/elagitannins. La fraction dialysable présente une réduction de 33% et de 35% de l'activité contre les radicaux dérivés de l’azote DPPH et NO, respectivement. De même, la capacité d'inhiber au niveau intracellulaire les radicaux libres (ROS) a diminué d’environ 85%. Toutefois, aucune différence significative n'a été mise en évidence au niveau de la capacité d'inhiber la peroxydation lipidique. Finalement, cette étude a utilisé un modèle in vivo pour évaluer l'effet de la consommation d'une boisson de mûre chez des rats diabétiques. L'apport de mûre a provoqué une diminution du glucose (-48.6%), des triglycérides (-43.5%) et du taux de cholestérol (- 28.6%) plasmatique des rats diabétiques par rapport aux témoins diabétiques qui n’ont pas consommé la boisson de ce fruit. De plus, les rats diabétiques qui consomment de la mûre ont vus améliorées les valeurs de la capacité antioxydante du plasma (+7%), tandis que les niveaux de peroxydation lipidique dans le plasma (-19%) et dans les tissus rénaux (-23%) ont été réduits. Cette étude suggère que la mûre pourrait être considérée comme un candidat pour le groupe des aliments fonctionnels, même si l'on considère les aspects de biodisponibilité et les effets du procédé de pasteurisation. Mots clés: Mûre, Rubus adenotrichos, activité anti-oxydante, activité anti-inflammatoire, diabète sucré, pasteurisation, biodisponibilité. ii Gratitude I thank Pr. Michel, Dr. Vaillant and Pr. Cristol for all the knowledge shared with me during all this doctoral processes. I thank Silvia Quesada, Marion Morena and Ana Mercedes Pérez for their scientific advice and even more for their friendship. I thank Karine Portet and Ana Lorena Torres for their excellent technical support. I thank all the staff of the Faculté de Pharmacie (UM1), IURC and CIRAD for all their technical advice and for making me feel welcome, particularly Frederic Boudard, Eric Badia, Christian Mertz, Florence Laurent, Patrick Poucheret, Jean Giaimis, Aurelie Goux, Nadine Lopez… I thank all my colleges on the Biochemistry department at University of Costa Rica for their support and friendship. I thank the University of Costa Rica, the Institute Francais d´Amerique Central and CIRAD for their financial support for my doctoral studies Last but not least, I thank my parents for all their support and motivation during my whole life, my sister for all her language support in the writing of this document and my brother for his prayers for me. I thank Carlos for his absolute support during all this process. Thanks for following me to France, for listening all the details of my experiments, for helping me in all the technological issues and for your unconditional support in all the aspects of my life. I thank God for all the doors He opened during this process and for giving me Alejandro, who is the inspiration of my life. iii Table of Contents Abstract ............................................................................................................................ i Résumé ............................................................................................................................. ii Table of Contents ........................................................................................................... iv Index of Tables ............................................................................................................. viii Index of Figures ............................................................................................................. ix List of Abbreviations ...................................................................................................... x Résumé ..........................................................................................................................
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