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Pdfs/ Endosulfan/Enviro Fate3.Pdf) ANNÉE 2014 THÈSE / UNIVERSITÉ DE RENNES 1 sous le sceau de l’Université Européenne de Bretagne pour le grade de DOCTEUR DE L’UNIVERSITÉ DE RENNES 1 Mention : Biologie et Sciences de la Santé École doctorale Vie-Agro-Santé présentée par Camille Savary Préparée à l’unité de recherche INSERM UMR 991 Foie, Métabolismes et Cancer (Pharmacie) Thèse soutenue à Rennes le 2 Juillet 2014 devant le jury composé de : Étude de la toxicité Pr François SICHEL Professeur des Universités chronique et du Université de Caen / rapporteur Dr Martine AGGERBECK potentiel cancérogène Chargé de recherche Paris Descartes, INSERM UMR-S 1124 / rapporteur de contaminants de Dr Brigitte LE MAGUERESSE Directeur de recherche l’environnement Université de Lyon, INSERM U1060 / examinateur Pr Lydie SPARFEL-BELIVET séparément et en Professeur des Universités Université de Rennes, IRSET / examinateur mélange sur les Pr André GUILLOUZO Professeur des Universités Université de Rennes 1, Inserm UMR 991 / directeur cellules HepaRG de thèse Dr Marie-Anne ROBIN Directeur de recherche Université de Rennes 1, Inserm UMR 991 / co- directeur de thèse Table des matières Index des Figures ............................................................................................................ 5 Index des tableaux .......................................................................................................... 7 Principales abréviations ................................................................................................... 8 Introduction générale ..................................................................................................... 13 I. Le foie, ses fonctions et les modèles d’études in vitro ............................................... 13 A. Le foie .................................................................................................................. 13 1. Les cellules hépatiques .................................................................................... 13 2. Le foie et ses principales fonctions ................................................................... 14 3. Les fonctions de détoxication du foie ................................................................ 16 B. Les modèles cellulaires hépatiques in vitro .......................................................... 30 1. Les différents modèles hépatiques in vitro ....................................................... 30 2. Le cas particulier de la lignée HepaRG ............................................................ 31 II. Les pesticides ............................................................................................................ 35 A. Avant-propos ....................................................................................................... 35 B. Les pesticides ...................................................................................................... 36 1. Les organophosphorés (OP) ............................................................................ 36 2. Les organochlorés (OC) ................................................................................... 37 C. Dispersion et devenir des pesticides dans l’environnement ............................. 39 1. Dispersion dans l’air, pollution atmosphérique ................................................. 39 2. Pollution des milieux aquatiques ...................................................................... 40 3. Migration et dégradation des pesticides dans les sols ...................................... 40 D. Les risques pour la santé humaine ................................................................... 41 1. Expositions ....................................................................................................... 41 2. Principaux effets des pesticides sur la santé humaine ..................................... 42 3. Mécanismes moléculaires liant pathologies chroniques et exposition aux pesticides ................................................................................................................ 47 E. Pesticides et la problématique des mélanges ...................................................... 51 1. La reconnaissance des risques liés aux mélanges de pesticides ..................... 51 2. L’approche expérimentale des effets de mélanges .......................................... 53 III. Cancérogenèse chimique ......................................................................................... 57 A. Généralité sur la transformation des cellules cancéreuses .................................. 57 1. Étapes de la cancérogénèse ............................................................................ 57 1 2. Propriétés des cellules tumorales ..................................................................... 58 B. Cancérogenèse chimique .................................................................................... 66 1. Les atteintes de l’ADN ...................................................................................... 66 2. Mécanismes de réparation de l’ADN ................................................................ 67 3. Les mécanismes épigénétiques ....................................................................... 68 4. Le stress oxydant et inflammation .................................................................... 69 C. Classification du risque carcinogène des composés chimiques ....................... 70 1. UE .................................................................................................................... 70 2. CIRC ................................................................................................................. 71 D. Les modèles utilisés pour la cancérogenèse chimique ..................................... 72 1. Les tests de génotoxicité .................................................................................. 72 2. Les tests in vitro de transformation cellulaire .................................................... 73 3. Les tests de cancérogenèse sur les rongeurs .................................................. 74 4. Études de cancérogenèse chimique à long terme in vitro ................................ 75 5. Recherche de biomarqueurs par les approches « omiques » .......................... 77 Objectifs de la thèse ...................................................................................................... 79 Chapitre 1 : Cellules HepaRG : évaluation fonctionnelle à confluence et stabilité au cours du temps .............................................................................................................. 81 I. Evaluation de la stabilité des cellules HepaRG : étude transcriptomique ................... 81 A. Article ................................................................................................................... 81 B. Résultats complémentaires : activités des CYPs dans les conditions de culture utilisées ...................................................................................................................... 83 II. Biocinétique de 4 médicaments après traitement unique ou répété dans les cellules HepaRG ......................................................................................................................... 84 A. Médicaments étudiés ........................................................................................... 84 1. Ciclosporine (CsA) ............................................................................................ 84 2. Chlorpromazine ................................................................................................ 85 3. Amiodarone ...................................................................................................... 86 4. Ibuprofène (IBU) ............................................................................................... 86 B. Matériel et méthodes ........................................................................................... 88 1. Le protocole expérimental ................................................................................ 88 2. Exposition aux médicaments ............................................................................ 89 3. La mesure analytique des produits et métabolites............................................ 90 C. Résultats .......................................................................................................... 91 1. Ciclosporine ...................................................................................................... 91 2 2. Chlorpromazine ................................................................................................ 93 3. Amiodarone ...................................................................................................... 94 4. Ibuprofène ........................................................................................................ 96 D. Discussion ........................................................................................................ 97 E. Conclusion ........................................................................................................... 99 Chapitre 2 : Effets des pesticides individuellement ou en mélange sur les cellules HepaRG après exposition aigüe ou répétée ................................................................ 101 I. Impact de l’isomalathion sur la cytotoxicité du malathion dans les cellules HepaRG 101 A. Résumé de l’étude ............................................................................................. 101
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