title: Enhanced expression of ATP-binding cassette transporter A1 in non-rafts decreases the sensitivity of vascular endothelial cells to Shiga toxin.
abstract: Shiga toxin (Stx) binds to globotriaosyl ceramide (Gb3) receptors on the surface of vascular endothelial cells, which is followed by Gb3-dependent endocytosis, and initiates a cascade leading to cell damage. The Gb3 receptor is localized in lipid rafts, in which cholesterol is tightly packed primarily with sphingolipids in a liquid-ordered state. Recent studies have indicated that phosphodiesterase (PDE) type 4 inhibitors enhance the expression of ATP-binding cassette 1 (ABCA1) which promotes cholesterol efflux from non-rafts at the plasma membrane. Here we report that rolipram, a PDE4 inhibitor, reduced the sensitivity to Stx2 of human umbilical vascular endothelial cells in association with increased apolipoproteinA-I (apoA-I)-mediated cholesterol efflux, and shift of some Gb3 molecules from lipid rafts into non-rafts. Although rolipram treatment did not reduce Gb3 content at the plasma membrane and Stx binding to whole cells of HUVECs, it reduced Stx2 endocytosis. Knockdown of ABCA1 by transfection with siRNA ABCA1 in vascular endothelial cells abrogated the protective effect of rolipram on Stx2-exposed cells. Our present results suggest that the expression level of ABCA1 protein is one of critical determinants of Stx sensitivity levels in vascular endothelial cells.
authors.affiliation: Department of Bacteriology, Nara Medical University, School of Medicine, Kashihara, Nara, Japan.
authors.forename: Nobutaka
authors.initials: N
authors.lastname: Higashi
authors.forename: Youko
authors.initials: Y
authors.lastname: Matsumura
authors.forename: Fumiko
authors.initials: F
authors.lastname: Mizuno
authors.forename: Kazuki
authors.initials: K
authors.lastname: Kasahara
authors.forename: Shigeki
authors.initials: S
authors.lastname: Sugiura
authors.forename: Keiichi
authors.initials: K
authors.lastname: Mikasa
authors.forename: Eiji
authors.initials: E
authors.lastname: Kita
mesh_terms: D064286:ATP Binding Cassette Transporter 1; D018528:ATP-Binding Cassette Transporters / Q000037:antagonists & inhibitors / Q000096:biosynthesis*; D016632:Apolipoprotein A-I / Q000378:metabolism; D017209:Apoptosis; D002462:Cell Membrane / Q000737:chemistry*; D002478:Cells, Cultured; D002784:Cholesterol / Q000378:metabolism; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D055785:Gene Knockdown Techniques; D006801:Humans; D021962:Membrane Microdomains / Q000378:metabolism; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D034741:RNA, Small Interfering / Q000378:metabolism; D020889:Rolipram / Q000494:pharmacology; D022621:Shiga Toxin / Q000378:metabolism / Q000633:toxicity*; D014281:Trihexosylceramides / Q000378:metabolism*
chemical_list: C577836:ABCA1 protein, human; D064286:ATP Binding Cassette Transporter 1; D018528:ATP-Binding Cassette Transporters; D016632:Apolipoprotein A-I; D010726:Phosphodiesterase Inhibitors; D034741:RNA, Small Interfering; D014281:Trihexosylceramides; C018549:globotriaosylceramide; D022621:Shiga Toxin; D002784:Cholesterol; D020889:Rolipram