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Etude Phytochimique Et Plantes Médicinales Pseudo-Scorodonia

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Etude Phytochimique Et Plantes Médicinales Pseudo-Scorodonia REPUBLIQUE ALGERIENNE DEMOCRATIQUE ET POPULAIRE MINISTERE DE L’ENSEIGNEMENT SUPERIEUR ET DE LA RECHERCHE SCIENTIFIQUE UNIVERSITE ABOU-BEKR-BELKAID TLEMCEN Faculté des Sciences de la Nature et de la Vie et Sciences de la Terre et de l’Univers Département de Biologie Laboratoire des Produits Naturels « LAPRONA » THÈSE Présentée par Mme BELARBI Karima ép. BOUTERFAS En vue de l’obtention du diplôme de Doctorat en Biologie Cellulaire et Biochimie Option : Produits Naturels, Aspects Nutritionnels et Activités Biologiques Thème Etude phytochimique et activités biologiques de deux plantes médicinales de la région de Tlemcen Teucrium pseudo-Scorodonia Desf. et Erica arborea L. Soutenue le : 12/05/2018 Devant le jury composé de : Président : Mr DJAZIRI Rabeh (Pr) Université de Tlemcen Directrice de thèse : Mme ATIK-BEKKARA Fawzia (Pr) Université de Tlemcen Co-directrice de thèse : Mme BEKHECHI Chahrazed (Pr) Université de Tlemcen Examinateurs : Mr DJEBLI Noureddine (Pr) Université de Mostaganem Mr BELLAHCEN Miloud (Pr) Centre universitaire d’Aïn Témouchent Année universitaire : 2017-2018 Remerciements Avant toute chose, je remercie ALLAH, le tout puissant de m’avoir donné la santé, la patience, la puissance et la volonté pour réaliser ce travail. J’ai eu la chance d’effectuer ce travail de recherche dans le laboratoire des « Produits Naturels » (LAPRONA) du département de Biologie, de la Faculté des Sciences de la Nature et de laVie, des Sciences de la Terre et de l’Univers (SNV/STU), de l’Université Abou Bekr Belkaïd de Tlemcen. Tout d’abord, je souhaite remercier très chaleureusement mon professeur et mon encadreur, Mme ATIK-BEKKARA Fawzia, Professeur à l’Université de Tlemcen et Directrice du laboratoire des Produits Naturels (LAPRONA) qui m’a fait l’honneur de réaliser ce travail sous sa direction. Je suis reconnaissante, Madame pour votre disponibilité, votre confiance, votre rigueur scientifique, vos conseils, vos encouragements et votre soutien dans les moments les plus difficiles qui m’ont permis de mener à bien ce travail. Je tiens aussi à adresser mes plus vifs remerciements à Mme BEKHECHI Chahrazed, Professeur à l’Université de Tlemcen, qui m’a fait l’honneur d’avoir accepté d’être mon Co- encadreur, recevez ici, chère Madame mes sentiments de gratitude pour votre aide, vos précieux conseils, votre compréhension et votre gentillesse. Je tiens également à exprimer à mes deux directrices, mes sincères remerciements pour les nombreuses heures investies dans la correction du présent travail. Aux membres de jury : Je tiens à remercier intensément Mr DJAZIRI Rabah, Professeur à l’Université de Tlemcen, d’avoir accepté de présider le jury de la soutenance, qu’il trouve ici toutes mes expressions respectueuses. Je veux exprimer mes sincères remerciements à Mr DJEBLI Noureddine, Professeur à l’Université de Mostaganem, pour avoir accepté de juger ce travail. Que Mr BELLAHCEN Miloud, Professeur au centre universitaire d’Aïn Témouchent, trouve ici l’expression de ma reconnaissance pour l’honneur qu’il ma fait d’avoir accepté de juger ce travail. J’exprime également mes remerciements et ma profonde reconnaissance à Mme BELYAGOUBI-BENHAMMOU Nabila, Maître de Conférences à l’Université de Tlemcen, pour son aide et ses encouragements. Un grand merci à Mr BENABADJI Noury, Professeur à l’Université de Tlemcen, pour l’aide qu’il nous a fourni pour l’identification des espèces végétales. De très précieux remerciements vont au Dr El-HACI Imad Abdelhamid qui n’a pas hésité de m’aider et de ne m’avoir jamais privé de son savoir. Je vous remercie pour votre disponibilité, vos encouragements et votre soutien dans les moments les plus difficiles. Mes remerciements s’adressent également à Melle BEDDOU Fawzia, Maître de Conférences à l’Université d’Alger, pour son amitié, sa gentillesse et ses encouragements. Je souhaite manifester mes vifs remerciements à Mme BENSAID Ilhem, pour sa vraie amitié, son aide, son soutien, son écoute, sa compréhension et ses encouragements dans les moments les plus difficiles. Je n’oublie pas de remercier vivement tous les doctorants et les membres du laboratoire des Produits Naturels (LAPRONA). Je vous remercie pour tous les bons moments passés au laboratoire. Je dédie ce manuscrit à mon cher regretté père (Allah yerhamou), à ma mère, mes sœurs et mes frères pour leur amour sans limites et leurs sacrifices déployés à mon égard. Enfin, les mots les plus simples étant les plus forts, j’adresse toute mon affection à mon mari et mes enfants Mohamed Khalil Allah et Fatima Zohra. Je vous remercie pour votre disponibilité, votre soutien moral et surtout votre patience, merci mes chers pour la joie que vous m’en donnez. Résumé Dans le présent travail, nous nous sommes intéressés à l’étude phytochimique et activités biologiques des extraits de deux plantes médicinales récoltés dans la région de Tlemcen : Teucrium pseudo-Scorodonia Desf. (Lamiaceae) et Erica arborea L. (Ericaceae). Le screening phytochimique réalisé sur les feuilles et les tiges de nos espèces nous a permis de mettre en évidence la présence des composés phénoliques, notamment, les flavonoïdes et les tanins. Les différents extraits organiques obtenus à partir des feuilles et des tiges de T. pseudo- Scorodonia ont présenté des teneurs considérables en polyphénols totaux et en flavonoïdes totaux. Les extraits bruts méthanoliques des feuilles ont présenté les teneurs les plus élevées. Ces teneurs sont de l’ordre de 266,36 ± 0,14 mg GAE/g, 161,23 ± 0,18 mg EC/g, respectivement. L’évaluation de l’activité antioxydante in vitro a révélé que tous les extraits présentent des capacités antioxydantes à différents niveaux. La fraction butanolique des feuilles a présenté la meilleure activité de piégeage du radical libre DPPH• (CI50 = 0,05 ± 0,00 mg/ml) et un fort pouvoir réducteur (CE50 = 0,19 ± 0,00 mg/ml). Ces valeurs sont inférieures à celles de la Vit E (CI50/DPPH = 0,07 ± 0,01 mg/ml ; CE50/FRAP = 0,24 ± 0,01 mg/ml). Les différents extraits ont été, également, évalué pour leur pouvoir antimicrobien. Cependant, aucune activité antimicrobienne n’a été révélée dans ces extraits. L’analyse chromatographique par RP-HPLC-PDA menée sur la fraction butanolique des feuilles a permis de mettre en évidence la présence de la rutine et de sept acides phénoliques (l’acide férulique, l’acide sinapique, l’acide rosmarinique, l’acide p-coumarique, l'acide caféique et l'acide syringique). Les différents extraits organiques obtenus à partir des feuilles et des tiges d’E. arborea ont exprimé des teneurs variables en polyphénols totaux et en flavonoïdes totaux. Les teneurs les plus élevées sont enregistré pour l’extrait brut méthanolique des feuilles à raison de 168,23 ± 3,54 mg GAE/g et 151,26 ± 3,65 mg EC/g, respectivement. La fraction butanolique des feuilles d’E. arborea a exprimé une valeur de CI50 de l’ordre de 0,03 ± 0,00 mg/ml pour le test de piégeage du radical libre DPPH•, comparable à celle de l’antioxydant de synthèse BHT. Pour le test de pouvoir réducteur, c’est la fraction tannique des feuilles qui a présenté le fort pouvoir réducteur avec une CE50 de l’ordre de 0,13 ± 0,00 mg/ml. En revanche, les extraits des feuilles et la fraction tannique des tiges montrent une certaine sensibilité contre Staphylococcus aureus avec des CMI et des CMB comprises entre 0,625 à 5 mg/ml. Six composés phénoliques ont été identifiés dans la fraction d’acétate d’éthyle des feuilles en utilisant la méthode RP-HPLC-PDA, il s’agit de : la rutine, l’acide p-coumarique, l'acide caféique, l’acide sinapique, l’acide férulique et l'acide syringique. Ces composés sont reconnus par leurs intéressantes propriétés thérapeutiques. Mots clés : Teucrium pseudo-Scorodonia, Erica arborea, Composés phénoliques, Activité antioxydante, Activité antimicrobienne, CCM, RP-HPLC-PDA. Abstract In the present work, we were interested to the phytochemical study and biological activities of the extracts of two medicinal plants collected from the region of Tlemcen; Teucrium pseudo- Scorodonia Desf. (Lamiaceae) and Erica arborea L. (Ericaceae). The phytochemical screening realized on the leaves and stems of our species allowed us to highlighting the presence of phenolic compounds, including, flavonoids and tanins. The different organic extracts obtained from the leaves and stems of T. pseudo-Scorodonia showed considerable contents of total polyphenols and total flavonoids. Methanolic crude extracts of the leaves showed the highest contents on the order of 266,36 ± 0,14 mg GAE/g and 161,23 ± 0,18 mg EC/g, respectively. The in vitro evaluation of the antioxidant activity has founding that all extracts showed antioxidant capacities at different levels. The butanolic fraction of the leaves exhibited the best DPPH• free radical scavenging activity (IC50 = 0,05 ± 0,00 mg/ml) and a high reducing power (EC50 = 0,19 ± 0,00 mg/ml), these values are lower than those of the Vit E (IC50/DPPH = 0,07 ± 0,01 mg/ml ; EC50/FRAP = 0,24 ± 0,01 mg/ml). The various extracts were also evaluated for their antimicrobial power, however, no antimicrobial activity was revealed in these extracts. The RP-HPLC-PDA chromatographic identification carried out on the butanolic fraction of the leaves allowed us to detect the presence of rutin and seven phenolic acids (ferulic acid, sinapic acid, rosmarinic acid; p-coumaric acid, caffeic acid and syringic acid). The different organic extracts obtained from the leaves and stems of E. arborea expressed variable contents of total polyphenols and total flavonoids, the highest contents reported in the methanolic crude extract of the leaves at 168,23 ± 3,54 mg GAE/g and 151,26 ± 3,65 mg EC/g, respectively. The butanolic fraction of the leaves of E. arborea expressed an IC50 value of 0,03 ± 0,00 mg/ml in the DPPH• free radical scavenging test, a value which is comparable to that of the synthetic antioxidant BHT, for the test of reducing power, it is the tannic fraction of the leaves which presented the strong reducing power with an EC50 on the order of 0,13 ± 0,00 mg/ml.
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