Lipid hydroperoxides inhibit nitric oxide production in RAW264.7 macrophages.
The effects of oxidatively modified low density lipoprotein (oxLDL) on atherogenesis may be partly mediated by alterations in the production of nitric oxide (NO) by vascular cells. Lipid hydroperoxides (LOOH) and lysophosphatidylcholine (lysoPC) are the major primary products of LDL oxidation. The purpose of this study was to characterize the effects of oxLDL, LOOH and lysoPC on NO production and the expression of inducible nitric oxide synthase (iNOS) gene in lipopolysaccharide (LPS) stimulated macrophages. LDL was oxidized using an azo-initiator 2,2'-azobis (2-amidinopropane) HCl (ABAP) and octadecadienoic acid was oxidized by lipoxygenase to generate 13-hydroperoxyl octadecadienoic acid (13-HPODE). Our study showed that oxLDL markedly decreased the production of NO, the levels of iNOS protein and iNOS mRNA in LPS stimulated macrophages. The inhibition potential of oxLDL on NO production and iNOS gene expression depended on the levels of LOOH formed in oxLDL and was not due to oxLDL cytotoxicity. Furthermore, 13-HPODE markedly reduced NO production and iNOS protein levels, whereas lysoPC showed only slight reduction. The effects of 13-HPODE and lysoPC did not require an acetylated LDL carrier. Our results suggest that 13-HPODE is a much more potent inhibitor of NO production and iNOS gene expression than lysoPC in LPS stimulated RAW264.7 macrophages.
Huang
A
A
Department of Pediatrics and Physiology, James H. Quillen College of Medicine, East Tennessee State University, Johnson City 37614-0578, USA.
Li
C
C
Kao
R L
RL
Stone
W L
WL
D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D006801:Humans; D007700:Kinetics; D008041:Linoleic Acids / Q000138:chemical synthesis / Q000494:pharmacology; D008054:Lipid Peroxides / Q000138:chemical synthesis / Q000494:pharmacology*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008077:Lipoproteins, LDL / Q000097:blood / Q000494:pharmacology*; D008244:Lysophosphatidylcholines / Q000494:pharmacology*; D008264:Macrophages / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D051379:Mice; D009569:Nitric Oxide / Q000096:biosynthesis*
D008041:Linoleic Acids; D008054:Lipid Peroxides; D008070:Lipopolysaccharides; D008077:Lipoproteins, LDL; D008244:Lysophosphatidylcholines; C099252:oxidized low density lipoprotein; C038649:13-hydroperoxy-9,11-octadecadienoic acid; D009569:Nitric Oxide