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Contribution À L'étude Phytochimique Et Recherche D'activités

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Contribution À L'étude Phytochimique Et Recherche D'activités République Algérienne Démocratique et Populaire Ministère de l’Enseignement Supérieur et de la Recherche Scientifique Université Aboubekr Belkaïd -Tlemcen- Faculté des Sciences de la Nature et de la Vie, des Sciences de la Terre et de l'Univers Département de Biologie Laboratoire Antibiotiques Antifongiques : Physico-chimie, Synthèse et Activité Biologique Thèse en vue de l’obtention du diplôme de Doctorat en Biologie Option : Substances à visée thérapeutique Présentée par : CHEKROUN Echaimaa Thème Contribution à l’étude phytochimique et recherche d’activités antioxydante et antidiabétique de deux cucurbitacées : Bryonia dioica Jacq et Citrullus colocynthis (L.) Schrad Devant le jury composé de : Président Dr. LAHFA Farid Université Aboubekr Belkaïd Tlemcen Directeur de thèse Pr. DJAZIRI Rabah Université Aboubekr Belkaïd Tlemcen Examinateurs Pr. BENAZZOUG Yasmina Université USTHB Alger Dr. BENMEHDI Houcine Université Mohammed Tahri Béchar Dr. AZZI Rachid Université Aboubekr Belkaïd Tlemcen Année universitaire : 2015-2016 Dédicaces Je dédie spécialement cette thèse à mes chers parents, pour leurs encouragements, leur tendresse, leur amour et leur soutien durant mes études ; vous trouverez ici le fruit de vos sacrifices et je souhaite que Dieu vous accorder une longue vie pleine de santé et de bonheur À ma grande mère À mes frères et ma petite sœur À mes nièces et mes neveux À toute ma famille et mes amis Echaimaa Remerciements J’exprime tout d’abord mes remerciements les plus sincères à mon directeur de thèse Mr DJAZIRI Rabah, Professeur au département de Biologie, faculté SNV/STU, Université Aboubekr Belkaïd Tlemcen, d’avoir accepté la charge de m’encadrer, pour son bienveillance au laboratoire et pour l’intérêt qu’il a donné pour accomplir ce travail. Je le remercie pour sa disponibilité, sa patience, ses conseils, sa compréhension, ses qualités pédagogiques et scientifiques. Veuillez trouver Monsieur, dans cette thèse, le témoignage de ma sincère reconnaissance et de mon profond respect. J’adresse mes sincères remerciements à Mr LAHFA Farid, Maître de conférences Classe A et Doyen de la faculté SNV/STU, Université Aboubekr Belkaïd Tlemcen, pour l’honneur qu’il m’a fait en acceptant de présider le jury de cette thèse, d’évaluer ce travail et de l’intérêt qu’il lui accordé. Je le remercie également pour sa compréhension et son aide précieuse à la réalisation de cette soutenance. J’exprime mes sincères remerciements à Mm BENAZZOUG Yasmina, Professeur à l’Université des Sciences et de la Technologie Houari Boumediène Alger, d’avoir accepté de participer à l’évaluation de ce travail et de l’enrichir de sa haute compétence scientifique. Je tiens à remercier Mr BENMEHDI Hocine, Maître de conférences Classe A à l’Université Mohammed Tahri Béchar, pour l’honneur qu’il m’a fait d'être examinateur de cette thèse. Qu’il trouve dans cette thèse le témoignage de mes sincères reconnaissances. Je suis très honoré que Mr AZZI Rachid, Maître de conférences Classe A au département de Biologie, faculté SNV/STU, Université Aboubekr Belkaïd Tlemcen, ait accepté d’examiner ce travail. Je le remercie vivement pour l’aide varié qu’il m’a donnée tout le long de la réalisation de ce travail, pour sa disponibilité et sa gentillesse. Qu’il trouve ici le témoignage de mon profond respect. Mes remerciements s’adressent à Mm BOUCHERIT-OTMANI Zahia, Professeur au département de Biologie, faculté SNV/STU, Université Aboubekr Belkaïd Tlemcen, et directrice du Laboratoire «Antibiotiques Antifongiques : Physico-chimie, Synthèse et Activité Biologique». Je remercie également Mm BENARIBA Nabila, Maître de conférences Classe B au département de Biologie, faculté SNV/STU, Université Aboubekr Belkaïd Tlemcen, pour sa disponibilité malgré ses nombreuses occupations, et de son aide précieuse pour les publications des résultats dans les revues scientifiques. Je la remercie pour ses conseils, ses encouragements, et son aide moral et scientifique. Qu’elle trouve dans cette thèse l’expression de mes remerciements les plus chaleureux. Au terme de plusieurs mois de travaux, je tiens à remercier Mm ABBAS-BECHIRI Asma, Melle ADIDA Houria, Mm BEKHTI-MEZERAI Rabiha, Melle DJAZIRI Fatima Zohra et Mm FELAHI-ZERIOUH Wafaa, Doctorantes aux Laboratoires de recherche LAPSAB et LAPRONA, pour leur aide dans la réalisation des essais in vivo et pour les discussions enrichissantes qu’elles ont attribué à ce travail. Je tiens aussi à remercier l’ensemble des doctorants ainsi que les techniciens du laboratoire «Antibiotiques Antifongiques : Physico-chimie, Synthèse et Activité Biologique», pour leur amitié, leur bonne humeur et leur aide. Je remercie tout particulièrement les membres de ma famille, pour leur présence constante, leur soutien et leur aide inestimable. Aussi, mes remerciements s’adressent à tous ceux qui de près ou de loin, ont contribué directement ou indirectement à la réalisation de ce modeste travail. Avant-propos Les travaux qui ont fait l’objet de cette présente thèse ont été réalisés dans le Laboratoire Antibiotiques Antifongiques : Physico-chimie, Synthèse et Activité Biologique (LAPSAB), Faculté des Sciences de la Nature et de la Vie, des Sciences de la Terre et de l’Univers, Université Aboubekr Belkaïd, Tlemcen - Algérie. Notre problématique s’inscrit dans un axe de recherche pluridisciplinaire qui concerne l’étude des activités biologiques et pharmacologiques des plantes médicinales utilisées pour le traitement du diabète sucré. L’approche ethnopharmacologique est adoptée afin de recenser et inventorier les plantes utilisées par la population diabétique en Algérie. Les plantes sélectionnées sont étudiées pour leur efficacité et leur toxicité chez différents modèles de diabète expérimental, induits par la streptozotocine. L’étude phytochimique du matériel végétal est également entreprise en vue de déterminer leur composition en phytoconstituants, de révéler les substances actives et d’élucider leur mécanisme d’action. الملخص نطبنًب سبْى انطب انخقهٛذ٘ كأسبط نهبحذ ػٍ أدٚٔت صذٚذة ػٍ غشٚق انفحص انخضشٚبٙ انكًٛٛبئٙ ٔانبٕٛنٕصٙ نًسخخهصبث انُببحبث انطبٛت. يٍ بٍٛ ْزِ انُببحبث Citrullus colocynthis (L.) Schrad ٔ Bryonia dioica Jacq ارُخٍٛ يٍ انقشٛػبث انًسخخذيت حقهٛذٚب نؼﻻس يشض انسكش٘، يشض يضيٍ خطٛش ٚخضاٚذ إَخشبسا فٙ صًٛغ أَحبء انؼبنى. انٓذف يٍ ْزِ اﻻغشٔحت ْٕ انذساست انكًٛٛبئٛت انُببحٛت نًسخخهصبث ْخٍٛ انُبخخٍٛ ٔحقٛٛى خصبئصًٓب انًعبدة نﻷكسذة فٙ انًخخبش ٔ انًعبدة نهسكش٘ فٙ انضسى انحػ ُٙذ انضشراٌ انًصببت ببنسكش٘ ػٍ غشٚق انحقٍ ببنسخشبخٕصٔحٕسػ (STZ) ٍُٛذ انٕﻻدة. حى ححعٛش يخخهف انًسخخهصبث يٍ صزٔس Bryonia dioica ٔ فٕاكّ Citrullus colocynthis : يسخخهصبث يبئٛت ٔ بٛخبَٕنٛت ببﻹظبفت انٗ يسخخهصبث انبٕنٛفُٕٛل ٔ انصببٍَٕٛ. كشفج انذساست انكًٛٛبئٛت انُببحٛت انٛػُٕت ٔانكًٛت نٓزِ انًسخخهصبث أسبسب ٔصٕد انكَُٕٛبث انحشة ٔانًشكببث انفُٕٛنٛت )انؼفص ٔانفﻻفَٕٛذاث( ٔانصببٍَٕٛ حشحٛشبٍٛ. ْزٍٚ اﻷخٛشاٌ ٕٚصذاٌ بكًٛبث يشحفؼت َسبٛب فٙ يسخخهصبث BDRSaponines ٔ BDRPolyphénols يغ ػ 952391 µg eq AO/mg ES ٔ 823326 µg eq AG/mg ESهٗ انخٕانٙ. دساست انُشبغ انًعبد نﻷكسذة ببسخخذاو غشٚقخٍٛ : غشٚقت انكسح انشادٚكبنٛت انًعبدة نﻷكسذة ػهٗ DPPH ٔ غشٚقت إسصبع انحذٚذ )FRAP(، أظٓشث َشبغ يعبد نﻷكسذة يخؼهق بخشكٛب انًسخخهص انًخخبش، حٛذ ٚحخٕ٘ انًسخخهص BDRPolyphénols (ػ (CI50=1,25 μg/mL ; CE50=21,67 µg/mLهٗ أْى َشبغ يعبد نﻷكسذة. ٔببﻹظبفت إنٗ رنك، فئٌ ْزا انًسخخهص بخشكٛض 1 يهغ / يم ﻻ ٚؤد٘ إنٗ إَحﻻل انذو غٕال 80 دقٛقت يٍ انحعبَت ػُذ 32 دسصت يئٕٚت. ْزٍٚ انُشبغٍٛ يعبد نﻷكسذة ٔ ٔاقٙ نهخهٛت يػ ٔ ًٍٛٓهٗ اﻷسصح ْزا ساصغ إنٗ إحخٕائًٓب ػهٗ يسخٕٚبث يشحفؼت يٍ انفُٕٛﻻث ٔانفﻻفَٕٛذاث انكهٛت. أظٓش حقٛٛى انخصبئص انًعبدة نهسكش٘ نضًٛغ يسخخهصبث C. colocynthisٔ B. dioica ، إَخفبظب قهٛﻻ فٙ يسخٕٖ انسكش فٙ انذو ػهٗ انًذٖ انقصٛش )3 سبػبث(، فٙ حٚ ٍٛصبح كبٛشا ػهٗ انًذٖ انًخٕسٚ 21( ػٕيب( ػُذيب حؼطٗ ٚ IPٕيٛب نهضشراٌ انًصببت بذاء انسكش٘. ببﻷخص، ٚخًٛض انًسخخهصبٌ (BDRSaponines (8 mg/kg pc) ٔ BDRn-but (40 mg/kg pc بخأرٛش كبٛش صذا فٙ خفط يسخٕٖ انسكش فٙ انذو ػهٗ انًذٖ انًخٕسػ ، فًٓب ٚعًُبٌ انحذ يٍ اسحفبع يسخٕٖ انسكش فٙ انذو بُسبت ػ ٪22- ٔ ٪86-هٗ انخٕانٚٔ ٙشصغ رنك أسبسب إنٗ احخٕائًٓب ػهٗ يسخٕٚبث يشحفؼت يٍ انصببٚٔ .ٍَٕٛشحبػ انًفؼٕل انًعبد نهسكش٘ ػهٗ انًذٖ انًخٕسػ بخحسٍٛ بؼط انًكَٕبث اﻷٚعٛت : انكٕنٛسخشٔل , انذٌْٕ انزﻻرٛت ٔانٕٛسٚب، ٔغٛشْب يٍ اﻻظطشاببث يزم انُٓبو، انؼطش ٔفقذاٌ انٕصٌ، كهٓب حغٛشاث ححذد بؼذ حقٍ ػ STZُذ انٕﻻدة. ػٔﻻٔة ػهٗ رنك، فئٌ انؼًبنضت بًسخخهصبث ْزِ انقشٛػبث نًذة ٚ 21ٕيب نى حظٓش أ٘ سًٛت فٙ انضشراٌ انطبٛؼٛت ٔ انًشٚعت ببنسكش٘. كم انُخبئش انخٙ حى انحصٕل ػهٛٓب فٙ ْزِ انذساست ٔاػذة، ﻷَٓب حخبشَب ػٍ انُشبغ انًعبد نﻷكسذة ٔ انًعبد نهسكش٘ نًسخخهصبث صزٔس B. dioica ٔ فٕاكّ C. colocynthis ٔ ْزا يب ٚؤكذ إنٗ حذ يب اسخخذايًٓب انخقهٛذ٘. ْزِ انخصبئص ْٙ أكزش أًْٛت ببنُسبت ل B. dioica. ْزِ انُخبئش ْٙ أٔنٛت ٔ حسخحق أٌ حؼًق أكزش يٍ أصم ححذٚذ ٔحٕصٛف انًشكببث انًسؤٔنت ػٍ ْزِ اﻷَشطت ٔيحبٔنت يؼشفت آنٛبث ػًهٓب ﻹدساصٓب فٙ انؼﻻس انذٔائٗ ظذ داء انسكش٘. الكلمات المفتاحية : Citrullus colocynthis ، Bryonia dioica ، انُشبغ انًعبد نﻷكسذة، انُشبغ انًعبد نًشض انسكش٘، انبٕنٛفُٕٛل، انصببٍَٕٛ، يشض انسكش٘ انخضشٚبٙ. Résumé La médecine traditionnelle a toujours servi comme une base à la recherche de nouveaux agents pharmacologiques actifs via le screening expérimental phytochimique et biologique des extraits de plantes médicinales. Parmi ces plantes Bryonia dioica Jacq et Citrullus colocynthis (L.) Schrad, deux cucurbitacées traditionnellement utilisées pour traiter le diabète sucré, maladie chronique grave dont la prévalence ne cesse d’augmenter à travers le monde. L’objectif de ce travail est consacré à l’étude phytochimique de quelques extraits et fractions de ces deux cucurbitacées ainsi que l’évaluation de leurs activités antioxydante in vitro et antidiabétique in vivo chez des rats rendus diabétiques par l’injection néonatale de la streptozotocine. Différents extraits aqueux et butanoliques en plus des fractions enrichies de polyphénols et de saponines ont été préparés à partir des racines broyées de B. dioica et des fruits concassés de C. colocynthis. L’étude phytochimique qualitative et quantitative de ces extraits a révélé principalement la présence de quinones libres, composés phénoliques (tanins et flavonoïdes) et de saponines triterpéniques. Ces deux derniers se trouvent à des taux relativement élevés dans les fractions BDRPolyphénols et BDRSaponines, avec des taux de 673,28 µg eq AG/mg ES et 592,51 µg eq AO/mg ES respectivement.
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