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Phytothérapie Et Polyphénols Naturels

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Phytothérapie Et Polyphénols Naturels REPUBLIQUE ALGERIENNE DEMOCRATIQUE ET POPULAIRE Ministère de l’Enseignement Supérieur et de la Recherche Scientifique Université Abdelhamid Ibn Badis Mostaganem Faculté Des Sciences De La Nature Et De La Vie Filière : Sciences Biologiques Spécialité : Microbiologie Appliquée Option : Interactions Micro- organismes, Hôtes et Environnements THÉSE PRESENTEE POUR L’OBTENTION DU DIPLOME DE DOCTORAT 3ème cycle LMD Par Mme. BENSLIMANE Sabria Contribution à l’étude de l’effet des extraits bruts des écorces du fruit de Punica granatum et des graines de Cuminum cyminum contre les biofilms à l’origine des infections bucco- dentaires. Soutenue le 18/01/2021 devant le jury: Président DJIBAOUI Rachid Pr Université de Mostaganem Directrice de thèse REBAI Ouafa MCA Université de Mostaganem Examinateur MEKHALDI Abdelkader Pr Université de Mostaganem Examinateur AIT SAADA Djamel MCA Université de Mostaganem Examinateur BEKADA Ahmed Med Ali Pr Centre Universitaire de Tissemsilet Année universitaire : 2020 -2021 Dédicaces Tout d’abord je tiens à dédier ce travail à la mémoire de ceux qui me sont chers mais qui ne font plus parti de ce monde, mon grand père Vladimir qui aurait été si fière de moi, mes grands parents paternel, ainsi que mon oncle parti si tôt, que dieu leur accorde sa miséricorde. À mes chers parents, pour tous leurs aides, leurs appuis, leurs dévouements, leurs sacrifices et leurs encouragements durant toutes mes années d’études. À ma très chère grand-mère Maria, qui m’a toujours soutenu et encouragé à poursuivre mes études, et pour tout ce qu’elle a fait pour moi depuis ma petite enfance. À mon très cher mari qui m’a toujours soutenu, encouragé et réconforté dans les moments les plus durs, merci pour ta compréhension et ton aide. À mes adorables enfants Mohamed Yanis et Lilia que dieu vous garde prés de moi et vous protège. À ma chère sœur Souria qui m’a toujours encouragé et aidé, à son mari Abdelghani, ainsi qu’à mes adorables neveux Abdelwaheb, Norhane et Lina Maria. À mes beaux parents, à mes belles sœurs Zineb et Nesrine, à mon beau frère Sofiane à mes neveux Nihel et Sadek Rayane ainsi qu’à la grand-mère Khdidja. À ma chère Ilhem pour son aide, à sa mère Samira et à ses filles adorée Kawtar, Mokhtaria et Rania. À mes amies que j’adore Asmaa, Kawtar, Katarina et Zuzanna pour vos encouragements et votre réconfort. À toute ma famille, mes oncles, mes tantes, mes cousins et cousines. I Remerciements En premier lieu, je remercie « Allah » le tout puissant, de m’avoir donné la force, le courage et la patience pour traverser ce long chemin difficile et enfin aboutir au bout de ce travail. Mes remerciements les plus vifs s’adressent à ma directrice de thèse Docteur REBAI Ouafa, pour ses conseils, son soutien, sa disponibilité et sa confiance tout au long de ce travail. J’exprime mes respectueux remerciements au professeur DJIBAOUI Rachid pour l’aide et les conseils qu’il a pu m’apporter, pour sa générosité et ses encouragements, ainsi que pour avoir accepté de présider le jury de soutenance. Mes remerciements vont également aux membres du jury pour avoir accepté d’examiner et d’évaluer ce travail. À monsieur MEKHALDI Abdelkader, professeur à l’université de Mostaganem et directeur du laboratoire de Microbiologie et Biologie Végétale, pour ses conseils, sa disponibilité et son soutien. À monsieur AIT SAADA Djamel Maitre de conférence A à l’université Abdelhamid Ibn Badis de Mostaganem et à monsieur BEKADA Ahmed Mohamed Ali Professeur à l’Université de Tissemsilt, de l’honneur qu’ils m’ont fait en acceptant de faire partie du jury. Ma sincère reconnaissance s’adresse à Monsieur ARABI Abed de l’université d’Oum El Bouaghi, pour ces précieux conseils, son aide, sa générosité ainsi que sa disponibilité tout au long de ce travail de thèse. Je tiens à exprimer ma sincère gratitude et ma reconnaissance au professeur DALACHE Fatiha qui a grandement contribué à ma formation durant mes années d’études, pour son aide, ces encouragements, sa générosité et ces précieux conseils, sans elle je ne serais pas là aujourd’hui. J’adresse un grand merci aux dentistes du centre de santé Izza Abdellah pour leur accueil chaleureux. Mes remerciements les plus distingués sont orientés éventuellement aux techniciens de laboratoires: Djahira, Amina et Nabil pour leur grande aide, leur disponibilité, leur gentillesse, et leurs rôles dans l’achèvement de ce travail. Je remercie tous mes anciens collègues Wafae A, Aicha L, Nadia B, Amina Z, Amina B, Hiba C, Imane B, Hakim H, Hicham D, ainsi que les enseignants du laboratoire de Microbiologie et biologie végétale pour les bons moments qu’on a pu partager ensemble, pour leurs conseils et leurs encouragements. À ma collègue B. Asmaa pour tous les moments de joies et de peines qu’on a pu partager. Mes remerciements vont aussi à l’égard de Monsieur BENCHEHIDA A et Mr DAHLOUM L pour leurs aides dans les analyses statistiques des données recueillies au cours de mon étude. Un grand merci à tous les professeurs de mon cursus universitaire à l’université de Mostaganem qui ont contribué à ce que je suis aujourd’hui, ainsi qu’à toute personne qui a contribué de prés ou de loin à la réalisation de ce travail. BENSlIMANE Sabria II Résumé Les communautés microbiennes buccales principalement organisées en biofilm, responsable du développement des caries et des maladies parodontales, présentent une résistance croissante aux antibiotiques, en plus, des inconvénients que présentent les produits utilisés pour réduire et inhiber la plaque dentaire. L'utilisation d'agents antimicrobiens d'origine végétale pourrait servir de traitement alternatif efficace contre les infections buccales. La présente étude visait à évaluer les activités antioxydante, antimicrobienne et antibiofilm des extraits d’écorces du fruit de Punica granatum et des graines de Cuminum cyminum contre des micro-organismes responsables d’infections bucco-dentaires. Les extraits ont été obtenus par macération avec trois différents solvants à raison de 70%: acétone, éthanol et le méthanol. Concernant la grenade, les teneurs totales en polyphénols et en flavonoïdes les plus élevées ont été obtenues avec l’extrait éthanolique 204.67± 15.26 mg EAG/g de matière sèche (MS), 67.67± 1.53 mg EQ/g MS respectivement. La teneur en tanin la plus élevée a été observée avec l’extrait acétonique 220± 17.32 mg EC/g MS. D’autre part, pour le cumin, la plus grande teneur en polyphénols, flavonoïdes et tanins, a été obtenue avec l’acétone 42 ± 3.46 mg EAG/g MS, 24.49± 1.24 mg EQ/g MS et 34.95± 8.71 mg EC/g MS respectivement. L’analyse chromatographique par HPLC menée sur les deux extraits des plantes a permis de caractériser les principaux composés phénoliques. La péduncalagine, la punigluconine et la punicalagine en tant qu'ellagitanine prédominante ont été identifiées pour l’extrait éthanolique d’écorces de grenade. Concernant l’extrait acétonique de C. cyminum, les résultats montrent la présence de l’acide p-coumarique, plusieurs flavonoïdes et plusieurs apigénines. L’évaluation in vitro, de l’activité antioxydante de nos différents extraits, a été réalisée par deux méthodes, à savoir le DPPH et FRAP. Pour le premier test les valeurs les plus élevées de l’activité de piégeage ont été observées avec l’extrait éthanolique de grenade (IC50= 220±14 µg/ml) et l’extrait acétonique de cumin (IC50= 940±50 µg/ml). L’évaluation du pouvoir antimicrobien des extraits par la méthode de diffusion des puits dans un milieu gélosé, a montré que l’ensemble possèdent un pouvoir antimicrobien sur toutes les souches Gram+ ainsi que sur les levures, en revanche, les bactéries Gram- se sont montrés moins sensibles envers les extraits de grenades et insensibles aux extraits de cumin. Les valeurs de concentration minimale inhibitrice (CMI) enregistrées par les extraits étudiés varient de 0.0125 à 100 mg/ml pour la grenade et de 12.5 à 200 mg/ml pour le cumin. En outre, dans l'activité antibiofilm, les extraits testés ont montré un pouvoir inhibiteur de la formation de biofilm, et ceci de manière dépendante de la concentration, contre les souches qui ont révélé une sensibilité envers ces extraits dans le test de l’activité antimicrobienne. Mots clés : Punica granatum, Cuminum cyminum, extraits phénoliques, biofilm buccal, activité antioxydante, activités antimicrobienne et antibiofilm. III Abstract The oral microbial communities mainly organized in biofilm, responsible for the development of cavities and periodontal diseases, present an increasing resistance to antibiotics. In addition, the disadvantages of products used to reduce and inhibit dental plaque. The use of antimicrobial agents derived from plants could serve as an effective alternative treatment against oral infections. The present study aimed to evaluate the antioxidant, antimicrobial and antibiofilm activities of extracts from the peels of fruit of Punica granatum and seeds of Cuminum cyminum against microorganisms responsible of oral infections. The extracts were obtained by maceration with three different solvents at a rate of 70%: acetone, ethanol and methanol. For pomegranate, the highest total polyphenols and flavonoids contents were obtained with the ethanolic extract 204.67±15.26 GAE/g dry weight (DW), 67.67± 1.53 mg QE/g DW respectively. The highest tannin content was observed with the acetone extract 220± 17.32 mg CE/g DW. On the other hand, for cumin, the greatest content of polyphenols, flavonoids and tannins, was obtained with acetone 42± 3.46 mg GAE/g DW, 24.49± 1.24 mg EQ/g DW and 34.95± 8.71 mg EC/g DW respectively. The chromatographic analysis by HPLC carried out on the two plant extracts allowed to characterize the main phenolics compounds.
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