title: Identification of Oxidized Phosphatidylinositols Present in OxLDL and Human Atherosclerotic Plaque.
abstract: Oxidized low-density lipoprotein (OxLDL) plays an important role in initiation and progression of atherosclerosis. Proatherogenic effects of OxLDL have been attributed to bioactive phospholipids generated during LDL oxidation. It is unknown what effect oxidation has on the phosphatidylinositol (PtdIns) molecules in LDL, even though PtdIns is 6% of the total LDL phospholipid pool. We sought to identify and quantitate oxidized phosphatidylinositol (OxPtdIns) species in OxLDL and human atherosclerotic plaque. Bovine liver PtdIns was subjected to non-enzymatic and lipoxygenase-catalyzed oxidation. Reversed-phase liquid chromatography with negative ESI-MS identified and confirmed compounds by fragmentation pattern analysis from which an OxPtdIns library was generated. Twenty-three OxPtdIns molecules were identified in copper-oxidized human LDL at 0, 6, 12, 24, 30, and 48 h, and in human atherosclerotic plaque. In OxLDL, OxPtdIns species containing aldehydes and carboxylates comprised 17.3 ± 0.1 and 0.9 ± 0.2%, respectively, of total OxPtdIns in OxLDL at 48 h. Hydroperoxides and isoprostanes at 24 h (68.5 ± 0.2 and 22.8 ± 0.2%) were significantly greater than 12 h (P < 0.01) without additional changes thereafter. Hydroxides decreased with increased oxidation achieving a minimum at 24 h (5.2 ± 0.3%). Human atherosclerotic plaques contained OxPtdIns species including aldehydes, carboxylates, hydroxides, hydroperoxides and isoprostanes, comprising 18.6 ± 4.7, 1.5 ± 0.7, 16.5 ± 7.4, 33.3 ± 1.1 and 30.2 ± 3.3% of total OxPtdIns compounds. This is the first identification of OxPtdIns molecules in human OxLDL and atherosclerotic plaque. With these novel molecules identified we can now investigate their potential role in atherosclerosis.
authors.affiliation: Institute of Cardiovascular Sciences, St. Boniface Hospital, Albrechtsen Research Centre, University of Manitoba, Winnipeg, MB, Canada.
authors.forename: Devin
authors.initials: D
authors.lastname: Hasanally
authors.affiliation: Institute of Cardiovascular Sciences, St. Boniface Hospital, Albrechtsen Research Centre, University of Manitoba, Winnipeg, MB, Canada.
authors.forename: Andrea
authors.initials: A
authors.lastname: Edel
authors.affiliation: Institute of Cardiovascular Sciences, St. Boniface Hospital, Albrechtsen Research Centre, University of Manitoba, Winnipeg, MB, Canada.
authors.forename: Rakesh
authors.initials: R
authors.lastname: Chaudhary
authors.affiliation: Institute of Cardiovascular Sciences, St. Boniface Hospital, Albrechtsen Research Centre, University of Manitoba, Winnipeg, MB, Canada. aravandi@sbgh.mb.ca.|Section of Cardiology, Department of Internal Medicine, Bergen Cardiac Care Centre, St. Boniface General Hospital, University of Manitoba, 409 Tache Avenue, Winnipeg, MB, R2H 2A6, Canada. aravandi@sbgh.mb.ca.
authors.forename: Amir
authors.initials: A
authors.lastname: Ravandi
mesh_terms: D000447:Aldehydes / Q000032:analysis; D000818:Animals; D002264:Carboxylic Acids / Q000032:analysis; D002417:Cattle; D056148:Chromatography, Reverse-Phase / Q000379:methods; D003300:Copper / Q000737:chemistry; D006801:Humans; D008077:Lipoproteins, LDL / Q000378:metabolism*; D008084:Lipoxygenase / Q000378:metabolism*; D008099:Liver / Q000378:metabolism*; D010716:Phosphatidylinositols / Q000032:analysis* / Q000737:chemistry; D058226:Plaque, Atherosclerotic / Q000737:chemistry*
chemical_list: D000447:Aldehydes; D002264:Carboxylic Acids; D008077:Lipoproteins, LDL; D010716:Phosphatidylinositols; C099252:oxidized low density lipoprotein; D003300:Copper; D008084:Lipoxygenase
keywords: Atheroma; Atherosclerosis; Lipoproteins; Mass spectrometry; OxLDL; Oxidized phospholipids; Phosphatidylinositol