Analysis of inflammatory gene induction by oxidized phospholipids in vivo by quantitative real-time RT-PCR in comparison with effects of LPS

Vascul Pharmacol. 2002 Apr;38(4):219-27. doi: 10.1016/s1537-1891(02)00172-6.

Abstract

Oxidized phospholipids are thought to play a role in the development of atherosclerosis and other chronic inflammatory processes. In this study, we analyzed the expression of inflammatory genes induced by oxidized L-alpha-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphorylcholin (OxPAPC) in vitro and in vivo using quantitative real-time reverse transcriptase-polymerase chain reaction (RT-PCR). Cultured human umbilical vein endothelial cells (HUVEC) and monocyte-like U937 cells were treated with OxPAPC or lipopolysaccharide (LPS) for 3 h. For in vivo studies, OxPAPC or LPS was injected intravenously into female C57Bl/6J mice and different tissues were isolated after 3 h. We found that both OxPAPC and LPS induced expression of early growth response factor 1 (EGR-1) and monocyte chemoattractant protein 1 (MCP-1) in HUVEC and of JE, the mouse homologue of MCP-1, in liver and heart. Interestingly, OxPAPC but not LPS increased expression of heme oxygenase 1 (HO-1) in U937 cells, HUVEC, aorta, heart, liver, and isolated blood cells. In contrast, E-selectin was selectively induced by LPS, but not by OxPAPC. Finally, OxPAPC-induced expression of HO-1 was blocked by a platelet-activating factor (PAF) receptor antagonist. We conclude that oxidized phospholipids are biologically active in vivo and exert a specific response inducing a pattern of genes that is different from that induced by LPS. In addition, we demonstrate that the quantitative real-time RT-PCR technology is a proper tool to investigate differential inflammatory gene induction in vivo.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms
  • Animals
  • Autoantigens / biosynthesis
  • Chemokine CCL2*
  • Culture Techniques
  • DNA, Complementary / biosynthesis
  • DNA, Complementary / genetics
  • DNA-Binding Proteins / biosynthesis
  • Early Growth Response Protein 1
  • Female
  • Gene Expression Regulation / drug effects*
  • Immediate-Early Proteins*
  • Inflammation / genetics*
  • Inflammation / pathology*
  • Injections, Intravenous
  • Lipopolysaccharides / pharmacology*
  • Liver / drug effects
  • Liver / metabolism
  • Mice
  • Mice, Inbred C57BL
  • Oxidation-Reduction
  • Phosphatidylcholines / pharmacology*
  • Phospholipids / administration & dosage
  • Phospholipids / pharmacology*
  • Platelet Membrane Glycoproteins / antagonists & inhibitors
  • RNA, Messenger / biosynthesis
  • RNA, Messenger / genetics
  • Receptors, Cell Surface / antagonists & inhibitors
  • Receptors, G-Protein-Coupled*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transcription Factors / biosynthesis
  • Transcriptional Activation
  • Up-Regulation / drug effects

Substances

  • Autoantigens
  • Ccl2 protein, mouse
  • Chemokine CCL2
  • DNA, Complementary
  • DNA-Binding Proteins
  • Early Growth Response Protein 1
  • Egr1 protein, mouse
  • Immediate-Early Proteins
  • Lipopolysaccharides
  • Phosphatidylcholines
  • Phospholipids
  • Platelet Membrane Glycoproteins
  • RNA, Messenger
  • Receptors, Cell Surface
  • Receptors, G-Protein-Coupled
  • Transcription Factors
  • oxidized-L-alpha-1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphorylcholine
  • platelet activating factor receptor