Les transporteurs membranaires jouent un rôle primordial dans la pharmacocinétique de médicaments mais aussi dans le transport de composés endogènes. La maîtrise de modèle in vitro permettant d’évaluer leur implication dans des interactions médicamenteuses, notamment au niveau hépatique, est donc primordiale. L’utilisation de ces modèles permettra aussi de déterminer l’impact potentiel de contaminants environnementaux, comme les particules diesel, sur l’expression fonctionnelle de transporteurs. Nos résultats démontrent la fiabilité de modèles hépatocytaires (hépatocytes et cellules d’hépatome HepaRG) en culture monocouche pour l’étude du transport membranaire de médicaments. Grâce à ces modèles, nous avons mis en évidence que des extraits de particules diesel (DEPe) peuvent modifier l’expression et/ou la fonction de transporteurs membranaires hépatiques, les Organic Anion Polypeptide Transporter (OATP) et les Multidrug Resistance associated Protein (MRP). Au niveau pulmonaire, les DEPe peuvent aussi augmenter l’expression du complexe LAT1/CD98hc, un complexe protéique de transport d’acides aminés souvent associé à de mauvais pronostics dans les cas de cancer du poumon. En conclusion, nos résultats mettent en évidence que les DEPe peuvent intervenir dans la régulation de l’activité et de l’expression de transporteurs membranaires tant au niveau hépatique qu’au niveau pulmonaire. / Membrane transporters play a major role in the pharmacokinetic of drugs and in the transport of endogenous compounds. The development of in vitro models for the study of their expression and activity is therefore important to consider, notably for analyzing their interactions with drugs or environmental contaminants such as diesel exhaust particles. Our results demonstrated the reliability of hepatocytic cells (hepatocytes and highly differentiated hepatoma HepaRG cells) in monolayer culture for the study of membrane transport of drugs. Using these models allowed us to demonstrate that extracts of diesel exhaust particles (DEPe) can alter the expression and / or function of major liver transporters such as organic anion transporting polypeptides (OATP) and Protein Multidrug associated Resistance (MRP). In lung cells, DEPe can increase the expression of complex LAT1 / CD98hc a protein complex, that is associated with poor prognosis in lung cancer. In conclusion, our results demonstrated that DEPe can regulate activity and expression of membrane transporters at hepatic and lung level.

Mechanistic aspects of carbonaceous aerosol toxicity were examined with respect to the ability of particles to catalyse reactive oxygen species-generating redox cycling reactions. To investigate the role of secondary organic material, we examined two systems. In the first, two-stroke engine exhaust particles were found to increase their ability to catalyse redox cycling in the presence of a reducing agent, dithiothreitol (DTT), when the exhaust was exposed to ozone. This occurred through deposition of redox-active secondary organic aerosol (SOA) onto the particle that was ten times more redox active per microgram than the primary engine particle. In the second system, naphthalene SOA formed highly redox active particles. Activity was strongly correlated to the amount of the 1,4- and 1,2-naphthoquinone measured in the particle phase. However, these species and the newly quantified naphthalene oxidation product 5-hydroxy-1,4-naphthoquinone accounted for only 30% of the observed DTT decay from the particles. Gas-particle partitioning coefficients suggest 1,4- and 1,2-naphthoquinone are not strong contributors to ambient particle redox activity at 25°C. However, a large number of redox active species are unidentified. Some of these may be highly oxidised products of sufficiently low vapour pressure to be atmospherically relevant. DTT activity of diesel particles was found to be high per unit mass. The activity was found to be associated with the insoluble fraction as filtration of the particles nearly eliminated DTT decay. Neither methanol nor dichloromethane extracts of diesel particles exhibited redox activity, indicating that the redox active species are associated with the black carbon portion of the particles. Examination of particle concentration techniques found that use of water condensation to grow and concentrate particles introduced a large organic artefact to the particles. Experiments with concentrated inorganic particles suggest that the source of this artefact is from irreversible uptake of water-soluble volatile organic compounds. Overall, carbonaceous redox active species can be thought of as a continuum from small, water-soluble species to redox active functionalities on elemental carbon backbones. In addition to clearly defined, quantifiable species, future research may need to consider examining broader chemical classes or redox-active chemical functionalities to overcome the inherent complexity of these constituents.

Mechanistic aspects of carbonaceous aerosol toxicity were examined with respect to the ability of particles to catalyse reactive oxygen species-generating redox cycling reactions. To investigate the role of secondary organic material, we examined two systems. In the first, two-stroke engine exhaust particles were found to increase their ability to catalyse redox cycling in the presence of a reducing agent, dithiothreitol (DTT), when the exhaust was exposed to ozone. This occurred through deposition of redox-active secondary organic aerosol (SOA) onto the particle that was ten times more redox active per microgram than the primary engine particle. In the second system, naphthalene SOA formed highly redox active particles. Activity was strongly correlated to the amount of the 1,4- and 1,2-naphthoquinone measured in the particle phase. However, these species and the newly quantified naphthalene oxidation product 5-hydroxy-1,4-naphthoquinone accounted for only 30% of the observed DTT decay from the particles. Gas-particle partitioning coefficients suggest 1,4- and 1,2-naphthoquinone are not strong contributors to ambient particle redox activity at 25°C. However, a large number of redox active species are unidentified. Some of these may be highly oxidised products of sufficiently low vapour pressure to be atmospherically relevant. DTT activity of diesel particles was found to be high per unit mass. The activity was found to be associated with the insoluble fraction as filtration of the particles nearly eliminated DTT decay. Neither methanol nor dichloromethane extracts of diesel particles exhibited redox activity, indicating that the redox active species are associated with the black carbon portion of the particles. Examination of particle concentration techniques found that use of water condensation to grow and concentrate particles introduced a large organic artefact to the particles. Experiments with concentrated inorganic particles suggest that the source of this artefact is from irreversible uptake of water-soluble volatile organic compounds. Overall, carbonaceous redox active species can be thought of as a continuum from small, water-soluble species to redox active functionalities on elemental carbon backbones. In addition to clearly defined, quantifiable species, future research may need to consider examining broader chemical classes or redox-active chemical functionalities to overcome the inherent complexity of these constituents.