Leukocytes utilize myeloperoxidase-generated nitrating intermediates as physiological catalysts for the generation of biologically active oxidized lipids and sterols in serum.
The initiation of lipid peroxidation and the concomitant formation of biologically active oxidized lipids and sterols is believed to play a central role in the pathogenesis of inflammatory and vascular disorders. Here we explore the role of neutrophil- and myeloperoxidase (MPO)-generated nitrating intermediates as a physiological catalyst for the initiation of lipid peroxidation and the formation of biologically active oxidized lipids and sterols. Activation of human neutrophils in media containing physiologically relevant levels of nitrite (NO(2)(-)), a major end product of nitric oxide (nitrogen monoxide, NO) metabolism, generated an oxidant capable of initiating peroxidation of lipids. Formation of hydroxy- and hydroperoxyoctadecadienoic acids [H(P)ODEs], hydroxy- and hydroperoxyeicosatetraenoic acids [H(P)ETEs], F(2)-isoprostanes, and a variety of oxysterols was confirmed using on-line reverse phase HPLC tandem mass spectrometry (LC/MS/MS). Lipid oxidation by neutrophils required cell activation and NO(2)(-), occurred in the presence of metal chelators and superoxide dismutase, and was inhibited by catalase, heme poisons, and free radical scavengers. LC/MS/MS studies demonstrated formation of additional biologically active lipid and sterol oxidation products known to be enriched in vascular lesions, such as 1-hexadecanoyl-2-oxovalaryl-sn-glycero-3-phosphocholine, which induces upregulation of endothelial cell adhesion and chemoattractant proteins, and 5-cholesten-3beta-ol 7beta-hydroperoxide, a potent cytotoxic oxysterol. In contrast to the oxidant formed during free metal ion-catalyzed reactions, the oxidant formed during MPO-catalyzed oxidation of NO(2)(-) readily promoted lipid peroxidation in the presence of serum constituents. Collectively, these results suggest that phagocytes may employ MPO-generated reactive nitrogen intermediates as a physiological pathway for initiating lipid peroxidation and forming biologically active lipid and sterol oxidation products in vivo.
Schmitt
D
D
Department of Cell Biology,VDepartment of Cardiology, Cleveland Clinic Foundation, Cleveland, Ohio 44195, USA.
Shen
Z
Z
Zhang
R
R
Colles
S M
SM
Wu
W
W
Salomon
R G
RG
Chen
Y
Y
Chisolm
G M
GM
Hazen
S L
SL
D000818:Animals; D002384:Catalysis; D002417:Cattle; D002478:Cells, Cultured; D018920:Coculture Techniques; D004730:Endothelium, Vascular / Q000166:cytology; D006801:Humans; D006861:Hydrogen Peroxide / Q000097:blood; D008051:Lipid Bilayers / Q000097:blood; D015227:Lipid Peroxidation; D008077:Lipoproteins, LDL / Q000097:blood / Q000502:physiology; D008670:Metals / Q000097:blood; D018375:Neutrophil Activation; D009504:Neutrophils / Q000378:metabolism* / Q000502:physiology; D009573:Nitrites / Q000097:blood*; D009584:Nitrogen / Q000097:blood; D010084:Oxidation-Reduction; D009195:Peroxidase / Q000097:blood* / Q000502:physiology; D013261:Sterols / Q000097:blood*; D013438:Sulfhydryl Compounds / Q000097:blood; D013481:Superoxides / Q000097:blood
D008051:Lipid Bilayers; D008077:Lipoproteins, LDL; D008670:Metals; D009573:Nitrites; D013261:Sterols; D013438:Sulfhydryl Compounds; D013481:Superoxides; D006861:Hydrogen Peroxide; D009195:Peroxidase; D009584:Nitrogen