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These De Doctorat En Sciences N° d’’ordre… … … .... ار اا ااط ا RÉPUBLIQUE ALGÉRIENNE DÉMOCRATIQUE ET POPULAIRE MINISTÈRE DE L’ENSEIGNEMENT SUPERIEUR ET DE LA RECHERCHE SCIENTIFIQUE UNIVERSITÉ DJILLALI LIABES DE SIDI BEL ABBES FACULTÉ DES SCIENCES DE LA NATURE ET DE LA VIE Département des Sciences de l’Environnement TTHHEESSEE DDEE DDOOCCTTOORRAATT EENN SSCCII EENNCCEESS PPPPrrrréééésssseeeennnnttttééééeeee ppppaaaarrrr :::: Melle : KECHAR Kheira SSSpppéééccciiiiiiaaalllllliiiiiitttééé :::: Biiollogiie OOOppptttiiiiiiooonnn :::: Biiodiiversiité Végétalle et Valloriisatiion Intiitullée Etude éthnobotanique et phytochimique de Ballota hirsuta Benth du mont de Tessala (Sidi Bel’Abbes). Soutenue lle …..…/….…/.....2016 Devant ll’’honorablle jjury composé de : Présiident de jjury : Dr MENADI Noureddine.……...…...…. ...(M.C. A, UDL/SBA) Examiinateurs : Dr OUABED Asmahane………..…............ (M.C.A, Univ. Tiaret) Dr OULBACHIR Karima….….......... …. (M.C.A, Univ. Tiaret) Dr BACHIR-BOUIADJRA S.E………… (M.C.A, UDL/ SBA) Diirecteur de thèse : Pr HELLAL Benchaben……..…...…..... (Professeur, Univ. Tiaret) Année universitaire 20 15 – 2016 REMERCIEMENTS En premier lieu, je remercie Allah tout puissant, qui m’a Guidé et facilité le chemin de la réussite. Je tiens à remercier bien sincèrement mon encadreur, Pr HELLAL Benchaben de toute la peine qu’elle s’est donnée pour l’accompagnement de mon travail. Mes vifs remerciements vont à Dr BENAHMED-DJILALI Adiba pour son aide et son soutien Je remercie Dr MENADINoureddine pour avoir voulu présider le jury. Je remercie également Dr OUABED Asmahane, Dr OULBACHIR Karima et Dr BACHIR-BOUIADJRA pour avoir bien voulu examiner ce travail. C’est avec une grande gratitude, que je remercie mes amis«Youcef, Mostapha » pour l’aide efficace qu’il m’a apportée, et tous ceux qui ont participé de prés ou de loin à l’élaboration de ce travail, notamment mes collègues « MELLAH Rabha, BENAOUM Zineb». ــــص ارا ا أ 2012 ل ال اي ل Ballota hirsuta ي س و أظت أن ات ا ه ا أوع اوم . أ إاء ا ت Folin-Ciocalteu و ا ب AlCl 3 و ا أظت أن أوراق Ballota hirsuta ة ات ا ال , ا ا ال ا وراق ا ا ي. ا ا ت ا ا ق و اف (CCM, HPLC) د ود ا ت أوراق Ballota hirsuta و ا و اطت ا. اط اد ت اوراق ات ا وط اد ة ، إن أ ت ا أظ ط إ دات ة، ا اـ ط ر ب 0,07 / / . أ ل و أرورم أز ه Staphylococcus aureu ط اط ول اراص ( 0,49±33,33 1± 12 ) . أــ درا اــط اد ري ل ت أرو ت ــ ن إراز ه ات ارـــ Staphylococcus aureus .Pseudomonasaeruginosa ات ا : Ballota hirsuta Benth، اط اد ة، اط اد ر، ا RÉSUMÉ L’enquête réalisée en 2012 dans la ville de Sidi Bel Abbes et la commune de Tessala (Algérie occidentale) a permis d’identifier 11 pathologies traitées par les différentes parties de la ballote hirsute ( Ballota hirsuta Benth). Les maladies les plus fréquemment traitées par la plante étudiée sont la contusion, les blessures et les douleursrhumatismales. L’espèce étudiée est employée, dans la majorité des cas, sous forme de cataplasme et en poudre. Les dosages quantitatifs des polyphénols totaux par la méthode de Folin-Ciocalteu, les flavonoïdes par la méthode d’AlCl 3et les tanins ont révélé la richesse desfeuillesde Ballotahirsuta en métabolites doséesdurant les quatre saisons. Ces résultatsexpliquent l’utilisation abondante de cet organe en médecine traditionnelle. Les méthodes chromatographiques (CCM, HPLC) ont révélé l’existence de certaines classes flavonoïques, issues des feuilles de Ballota hirsuta , responsables de l’activité biologique. L’expression des résultats de l’activité anti-oxydante et antibactérienne a montré quel’extrait acétate d’éthyle est plus actif comme agents antioxydants avec une concentration inhibitrice de 50%de radicale libre DPPH est de l’ordre de 0,07 mg/ml .Cependant la fraction n- butanolique et chloroformique se sont montréesplus active contre Staphylococcus aureus avecdes diamètres d’inhibitions de 12 ±1à 33,33 ±0,49 mm. Les pigments chlorophylliens commela xanthophylle,produit terpénique, s’estavéré la plus active sur les trois souches Escherichia coli, StaphylococcusaureusetPseudomonasaeruginosa. Mots clés :Ballotahirsuta Benth , Activité Anti-oxydante, Activité antimicrobienne, Flavonoïdes. ABSTRACT The investigation conducted in 2012 in the city of SidiBel Abbes and the common of Tessala (western Algeria) allowed identify 11 diseases processed by the various parties of the shaggy horehound ( Ballota hirsute Benth). The diseases most frequently treated by the plant studied the contusion, the injuries and rheumatic pains. The species studied is employed, in most cases,as a poultice and powder. The quantitative assays of the total polyphenols by the method of Folin-Ciocalteu, the flavonoids by AlCl3 method and the tannins revealed the wealth of the leaves of the Ballota hirsute in metabolite measured during the four seasons. These results explain the abundant use of this organ in the traditional medicine. The Chromatographic methods (CCM, HPLC) revealed the existence of some flavonoïquesclasses, the outcome of the leaves of the Ballotahirsuta, responsible for the biological activity The expression of the results of the antioxidant and the antibacterial activity showed that the extract acétated’éthyle is more active like antioxidants agents with an inhibitory concentration of the 50% of free radical DPPH is of the order of 0.07 mg / ml. However the fraction n- butanol and chloroform have been more active with a view to Staphylococcus aureus with diameters of inhibition of 12 ±1 à 33,33 ±0,49 mm. The chlorophyll pigments as the xanthophyll, product the terpene, proved the more active of the three strains Escherichia coli, Staphylococcusaureus et Pseudomonas aeruginosa. Keywords :Ballotahirsuta Benth ,Antioxidantactivity,Antimicrobialactivity,Flavonoids. LISTE DES ABRÉVIATIONS CCM : Chromatographie sur couche mince DMSO : Diméthylsulfoxyde DO : Densité optique HPLC : Chromatographie en phase liquide à haute performance MEB :Microscope électronique à balayage AcOEt : Acétate d’éthyle cm : Centimètre MeOH : Méthanol mg : Milligramme min : Minutes ml : Millilitre DPPH : 2.2 diphenyl 1 picrylhydrazyl nm : Nanomètre Rf : Rapport frontal UV : Ultra-violet LISTE DES TABLEAUX Tableau n°1 : Pluviométrie moyennes de la zone d’étude (mm). ............................................ 17 Tableau n°2 : Répartition des précipitations annuelles (en mm) de la commune de Tessala (1985-2013). ............................................................................................................................. 18 Tableau n°3: Répartition saisonnière des précipitations dans la commune de Tessala(ONM,2014) ................................................................................................................. 21 Tableau n°4 : Moyenne mensuelle du taux d’humidité relative durant la période (1980/2009). .................................................................................................................................................. 21 Tableau n°5: Fréquence moyenne mensuelle des gelées Période (1985/2005)........................ 22 Tableau n°6 : Répartition général des classes de pentes .......................................................... 23 Tableau n°7 : Division des flavonoïdes .................................................................................... 35 Tableau n°8: Les différents systèmes solvants utilisés pour la CCM de gel de silice ............. 55 Tableau n°9 :germes étudies .................................................................................................... 58 Tableau n°10 : Les étapes de la déshydratation ....................................................................... 64 Tableau n°11 : Résultats de l’enquête ethnobotanique menée sur l’utilisation de la ballote hirsute( Ballota hirsuta Benth.) ................................................................................................. 74 Tableau n°12 : Caractères physico-chimiques de la matière végétale de la ballote hirsute du mont de Tessala (Algérie occidentale). .................................................................................... 76 Tableau n°13: Composition minérale de la matière végétale de la ballote hirsute du mont de Tessala (Algérie occidentale). .................................................................................................. 77 Tableau n°14: Résultats de screening dans les différentes organes de Ballota hirsuta Benth du mont de Tessala (Algérie occidentale). ............................................................................... 84 Tableau n°15: Concentration moyennes en métabolites des différents organes de la ballote hirsute du mont de Tesssala (Algérie occidentale)..................................................................85 Tableau n°16: Comportement chromatographique des flavonoïdes dans le système solvant 1 .................................................................................................................................................. 92 Tableau n° 17 : Les résultats de la chromatographie sur couche mince des témoins par le système solvant 2 ..................................................................................................................... 92 Tableau n°18: Comportement chromatographique dans le système solvant 2......................... 93 Tableau n°19 : Comportement chromatographique dans le système solvant 3........................ 94 Tableau n°20: Temps de rétention des flavonoïdes standard ................................................. 97 Tableau n°21 : Temps de rétention des flavonoïdes présents dans les extraits
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