title: Inhibition of cellular efflux pumps involved in multi xenobiotic resistance (MXR) in echinoid larvae as a possible mode of action for increased ecotoxicological risk of mixtures.
abstract: In marine organisms the multi xenobiotic resistance (MXR) mechanism via e.g. P-glycoprotein (P-gp) and multidrug resistance-associated protein (MRP) is an important first line of defense against contaminants by pumping contaminants out of the cells. If compounds would impair the MXR mechanism, this could result in increased intracellular levels of other compounds, thereby potentiating their toxicity. A calcein-AM based larval cellular efflux pump inhibition assay (CEPIA) was developed for echinoid (Psammechinus miliaris) larvae and applied for several contaminants. The larval CEPIA revealed that triclosan (TCS) and the nanoparticles P-85(®) (P-85) were 124 and 155× more potent inhibitors (IC(50) 0.5 ± 0.05 and 0.4 ± 0.1 μM, respectively) of efflux pumps than the model inhibitor Verapamil (VER). PFOS (heptadecafluorooctane sulfonic acid) and pentachlorophenol also were more potent than VER, 24 and 5×, respectively. Bisphenol A and o,p'-dichlorodiphenyltrichloroethane (o,p'-DDT) inhibited efflux pumps with a potency 3× greater than VER. In a 48 h early life stage bioassay with P. miliaris, exposure to a non-lethal concentration of the inhibitors TCS, VER, the model MRP inhibitor MK-571, the nanoparticles P-85 and the model P-gp inhibitor PSC-833, increased the toxicity of the toxic model substrate for efflux pumps vinblastine by a factor of 2, 4, 4, 8 and 16, respectively. Our findings show that several contaminants accumulating in the marine environment inhibit cellular efflux pumps, which could potentiate toxic effects of efflux pumps substrates.
authors.affiliation: Sub-department of Toxicology, Wageningen University, Tuinlaan 5, 6703 HE, Wageningen, The Netherlands.
authors.forename: Henrique M R
authors.initials: HM
authors.lastname: Anselmo
authors.forename: Johannes H J
authors.initials: JH
authors.lastname: van den Berg
authors.forename: Ivonne M C M
authors.initials: IM
authors.lastname: Rietjens
authors.forename: Albertinka J
authors.initials: AJ
authors.lastname: Murk
mesh_terms: D000818:Animals; D004305:Dose-Response Relationship, Drug; D018432:Drug Resistance, Multiple; D005452:Fluoresceins / Q000378:metabolism*; D007814:Larva / Q000187:drug effects / Q000254:growth & development / Q000502:physiology; D008856:Microscopy, Fluorescence; D012617:Sea Urchins / Q000187:drug effects* / Q000196:embryology / Q000254:growth & development / Q000502:physiology; D023382:Toxicity Tests, Acute / Q000379:methods*; D015262:Xenobiotics / Q000633:toxicity*
chemical_list: D005452:Fluoresceins; D015262:Xenobiotics; C007740:fluorexon