Inhibition of P-glycoprotein enhances sensitivity of Caenorhabditis elegans to ivermectin.

摘要

In vertebrates, the function of P-glycoprotein (PGP) is to protect against toxic compounds through active efflux of the toxin from target tissues. In clinical oncology, the overexpression of PGP confers drug resistance. The function(s) of PGP in nematode physiology or in conferring drug resistance is less understood. The objective of this study was to determine the role of PGP in drug resistance in nematodes using Caenorhabditis elegans and ivermectin (IVM) as the model system. The IVM sensitive wild-type Bristol N2 strain, seven PGP deletion strains and a triple IVM receptor (avr-14/avr-15/glc-1) knock-out strain showing synthetic resistance to IVM (IVM-R) were used to (1) compare the gene expression signatures of 15 PGPs in the wild-type and resistant strains following treatment; (2) measure motility and pharyngeal pumping phenotypes in the wild-type, IVM-R and PGP deletion strains before and after treatment; and (3) quantify the phenotypic responses of the wild-type and IVM-R strains to IVM or IVM co-administered with 12 chemosensitizers that interfere with PGP function. IVM induced changes in both amplitude and timing of gene expression for the 15 PGP genes. Following IVM treatment, the most significant effects were observed in the IVM-R strain for those PGP genes expressed in the neurons, pharynx and intestine. Inactivation of pgp-2, pgp-5, pgp-6, pgp-7, pgp-12 and pgp-13 resulted in increased sensitivity to IVM compared with the wild-type. The phenotypic responses of the IVM-R strain differed from those of the wild-type strain when exposed to IVM alone, or IVM co-administered with chemosensitizers. The phenotypic responses to the co-administration of chemosensitizers varied with the concentration of IVM used, suggesting that the action of PGP's is influenced by the concentration of IVM. Verapamil restored sensitivity to IVM in the IVM-R strain. Our results demonstrate that PGPs play a role in protecting C. elegans from IVM toxicity and inhibition of PGP enhances susceptibility to IVM. PGP may be a mechanism for multidrug resistance (MDR) in parasitic nematodes.