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Eryngium Campestre L. Et Paronychia Capitata L. Présentée Par : BOUZIDI Soumia

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Eryngium Campestre L. Et Paronychia Capitata L. Présentée Par : BOUZIDI Soumia REPUBLIQUE ALGERIENNE DEMOCRATIQUE ET POPULAIRE MINISTERE DE L’ENSEIGNEMENT SUPERIEUR ET DE LA RECHERCHE SCIENTIFIQUE UNIVERSITE MOSTEF ABEN BOULAÏD-BATNA 2 FACULTE DES SCIENCES DE LA NATURE ET DE LA VIE DEPARTEMENT DE BIOLOGIE DES ORGANISMES Thèse en vue de l’obtention du diplôme de DOCTORAT troisième cycle Filière : Sciences Biologique Option : Biotechnologie Des Molécules Bioactives Et Pathologie Moléculaire Thème Evaluation des activités biologiques des extraits de deux plantes : Eryngium campestre L. et Paronychia capitata L. Présentée par : BOUZIDI Soumia Devant le jury Président : LAROUI Salah Pr Université Batna 2 Examinateur : YAHIA Abdelouahab Pr Centre Universitaire Mila Examinateur : YAHIA Mouloud Pr Université Batna 2 Examinatrice : HAMBABA Leila Pr Université Batna 2 Examinateur : BOUFENNARA Souhil MCA Université de Khenchela Directrice de thèse : BENKIKI Naima MCA Université Batna 2 Année universitaire : 2017-2018 REPUBLIQUE ALGERIENNE DEMOCRATIQUE ET POPULAIRE MINISTERE DE L’ENSEIGNEMENT SUPERIEUR ET DE LA RECHERCHE SCIENTIFIQUE UNIVERSITE MOSTEF ABEN BOULAÏD-BATNA 2 FACULTE DES SCIENCES DE LA NATURE ET DE LA VIE DEPARTEMENT DE BIOLOGIE DES ORGANISMES Thèse en vue de l’obtention du diplôme de DOCTORAT troisième cycle Filière : Sciences Biologique Option : Biotechnologie Des Molécules Bioactives Et Pathologie Moléculaire Thème Evaluation des activités biologiques des extraits de deux plantes : Eryngium campestre L. et Paronychia capitata L. Présentée par : BOUZIDI Soumia Devant le jury Président : LAROUI Salah Pr Université Batna 2 Examinateur : YAHIA Abdelouhab Pr Centre Universitaire Mila Examinateur : YAHIA Mouloud Pr Université Batna 2 Examinatrice : HAMBABA Leila Pr Université Batna 2 Examinateur : BOUFENNARA Souhil MCA Université de Khenchela Directrice de thèse : BENKIKI Naima MCA Université Batna 2 Année universitaire : 2017-2018 In life, you will realize there is a role for everyone you meet… Some will test you, some will use you, some will love you and some will teach you. However, the ones who are truly important are the ones who will bring out the best in you. They are rare and amazing people who remind you “why it’s all worth it …” Remerciements Je remercie tout d’abord ALLAH, le Tout Puissant de m’avoir accordé santé, courage et foi. Pour L’aboutissement de ce travail de thèse est évidemment lié à des moments durant lesquels diverses contributions m’ont été bénéfiques. Je remercie, Madame BENKIKI NAIMA, Maitre de conférences à l'université Mustapha Ben Boulaid, Batna d’avoir dirigée mes travaux au cours de toutes ces années avec beaucoup de patience. Votre rigueur, vos conseils et vos encouragements m’ont permis de mener à bien ce travail. Je tiens à remercier sincèrement l’ensemble des membres du jury pour avoir accepté de juger ce travail, notamment : Monsieur le professeur LAROUI SALAH, l’Université Mustapha Ben Boulaid, Batna de me faire l’honneur de présider le jury de ma thèse. Messieurs les professeurs YAHIA MOULOUD (Université Mustapha Ben Boulaid, Batna) et YAHIA ABDELOUHAB (Centre Universitaire Mila) je suis honorée que vous ayez accepté d’être juges de ma thèse. Vos compétences étaient d’une valeur inestimable pour enrichir ce travail. Monsieur BOUFENNARA SOUHIL, Maitre de conférences à l'université Abbès Laghrour, Khenchela et Madame HAMBABA LEILA Professeur à l’Université Mustapha Ben Boulaid, Batna qui m’ont fait l’honneur de juger et siéger dans le jury de ma thèse. Une grande part de ma reconnaissance s'adresse à Monsieur le Professeur HABA HAMADA, l’Université Hadj Lakhder, Batna pour son aide précieuse et son soutien. Je tiens aussi à exprimer toute ma reconnaissance à Monsieur KALLA ADEL et à mademoiselle GOUGIL RIMA pour l'accueil et la disponibilité dont vous avez fait preuve dans votre laboratoire. Je vous remercie pour tous vos efforts, pour votre collaboration dans le suivi de l’activité antimicrobienne des plantes étudiées. Docteur ZINA ALLOUAI et Madame BOUZID WAFA, je vous exprime mes plus sincères remerciements pour votre présence, votre aide et votre gentillesse au quotidien, tous vos encouragements et votre soutien. Monsieur HACHEMI MASOUAAD je tiens à vous remercier chaleureusement pour votre soutien infaillible. Je vous exprime ici ma plus profonde reconnaissance pour vos conseils et votre aide concernant l’étude in vivo de ce travail. Mes sincères remerciements, à tous les membres de l’équipe du laboratoire de biotechnologie des molécules bioactives et de la physiopathologie cellulaire à l’université Batna 2. A mes collègues de l’université Mustapha Ben Boulaid, Batna en particulier, ceux et celles de la faculté des sciences de la nature et de la vie. Ainsi que mes collègues du staff administratif de la faculté des sciences de la nature et de la vie. Je ne saurais oublier de vous remercier, vous, OUMAIMA, MERIEM et BATOUL, chère sœurs, pour votre soutien, votre aide, votre présence ainsi que votre affection. Un très grand merci à mes parents pour leur soutien inconditionnel tout au long de mes études et la confiance qu’ils m’ont toujours témoignée. Merci à toute ma famille et mes amis pour leur présence à mes côtés. Aussi, un grand merci pour toute personne ayant contribué de près ou de loin à la réalisation de ce modeste travail. Abstract The main purpose of this work was the valorization of medicinal flora of Algeria, to promote the sustainable use of medicinal plant resources. This thesis specially aimed to evaluate the potential of two plants Eryngium campestre L. (Apiaceae) used as a folk remedy worldwide for the treatment of various disorders and Paronychia capitata L. (Caryophyllaceae) in the treatment of oxidative stress, microbes and inflammation related diseases. Moreover, this study was accomplished to evaluate and compare the in vitro antioxidant and antimicrobial activities of extracts obtained from the roots and aerial parts of Eryngium campestre L. and of various extracts of Paronychia capitata L. In addition, to assess the acute oral toxicity the in vivo antipyretic and the anti-inflammatory activities of the n-butanol extract of roots and aerial parts of Algerian Eryngium campestre L. and the n-butanol extract of Paronychia capitata L. Extracts were obtained of roots and aerial parts of E. campestre L. using several solvents: ethyl acetate, n-butanol and water the yields were: [ECRAC 0.82 %, ECAAC 0.71 % (w/w)], [ECRBU 4.08%, ECABU 3.05% (w/w)] and [ECRAQ 1.68, ECAAQ 1.55% (w/w)] respectively. Total phenolic contents were determined using Folin-Ciocalteu reagent and found to be 22.30, 27.77, 7.18, 21.52 and 18.21, 21.50 µg GAE/mg of extract in ethyl acetate, n-butanol and water (roots and aerial parts) extracts respectively. Flavonoids were evaluated by AlCl3 method and shown to be 3.58, 6.46, 3.21, 3.26 and 9.46, 7.54 µg QE/mg of extract in ethyl acetate, n-butanol and water (roots and aerial parts) extracts respectively. On one hand, organic extracts of P. capitata L. were obtained with three solvents: petroleum ether then ethyl acetate and final n-butanol. The yields were 0.41% (w/w), 0.78 % (w/w) and 3 % (w/w) respectively. Whereas the aqueous extract had a yield 0.69 % (w/w) of extraction. Moreover, the butanol extract of P. capitata L. PCBU contains a high amount of total polyphenols (32.23 µg GAE/mg of extract) and the highest flavonoids goes to the aqueous extract (6.08 µg QE/mg of extract) The antioxidant properties were evaluated using four different methods (CAT/FRAP / β-Carotene linoleic acid/ DPPH) of various extracts from roots, aerial parts of Eryngium campestre L. and from P. capitata L. According to the results of our investigation, butanol extract of aerial parts ECABU had considerably higher antioxidant activity in (CAT, FRAP and DPPH) assays (263.40 µg/mg extract, EC50 = 263.37µg/ml and IC50 value of 16.14µg/ml) while the roots aqueous extract ECRAQ showed the highest inhibition of oxidation in the β-Carotene linoleic acid with (89.78%). Furthermore, results indicated that n-butanol extract of P. capitata L. showed greater antioxidant activity than other P. capitata L. extracts in (CAT,DPPH and β-Carotene linoleic acid) assays (259.70 µg/mg extract, IC50 = 08.667µg/ml and 84.92 %). The in vitro screening of the antimicrobial activity was determined using eight bacterial strains (GRAM+ and GRAM-) and seven fungi strains, according to the disk diffusion assay; extracts of both plants have shown a modest inhibitory effect against the microorganisms tested. The acute oral toxicity study of butanol extracts of E. campestre L. (roots, aerial parts) and P. capitata L. were evaluated on Albino Wistar female rats. Extracts at a single dose of 2000 mg/kg did not produce treatment related signs of toxicity or mortality in any of the animals tested during 14 days observation period; therefore, the LD50 of both plants was estimated above 2000 mg/kg. The obtained results revealed no significantly difference (p< 0.05) in the hematological parameter, all the serum biochemical parameters when compared to the control group. Furthermore, the anti-inflammatory and antipyretic activities of the n-butanol extracts of both plants were investigated on Wistar albino rats at the dose levels of 250 and 500 mg/kg body weight, using the egg albumin induced oedema, and brewer’s yeast induced pyrexia method respectively. The n-butanol extracts of E. campestre L. showed a good anti-inflammatory activity comparable to the standard drug diclofenac and the butanol extract of roots ECRBU possessed a better antipyretic effects (at both dose 250 mg/kg and 500 mg/kg) compared to the butanol extract of aerial parts. Regarding the second plant P. capitata L., the n-butanol fraction evoked a non-dose related anti-inflammatory activity and inhibited significantly the increase of paw edema from 30 minutes to 5 hours.
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