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Lipoxin generation by permeabilized human platelets.

M Romano1, C N Serhan

  • 1Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115.

Biochemistry
|September 8, 1992
PubMed
Summary
This summary is machine-generated.

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Human platelets efficiently convert leukotriene A4 into lipoxins, especially when cell membranes are compromised. This study reveals lipoxin generation as a key metabolic pathway in activated platelets.

Area of Science:

  • Biochemistry
  • Cell Biology
  • Immunology

Background:

  • Human platelets are known to metabolize leukotriene A4 (LTA4).
  • Transcellular biosynthesis involves the conversion of LTA4 to lipoxins by platelets.

Purpose of the Study:

  • To investigate and compare lipoxin generation in intact versus permeabilized human platelets.
  • To characterize the lipoxins produced and the kinetics of their formation.

Main Methods:

  • Comparison of lipoxin and peptidoleukotriene generation in intact and permeabilized human platelets stimulated with thrombin.
  • Identification and quantification of lipoxins (LXA4, LXB4) and their isomers using High-Performance Liquid Chromatography (HPLC) and Gas Chromatography-Mass Spectrometry (GC-MS).
  • Chiral analysis of lipoxin stereochemistry and kinetic studies (Km, Vmax) under varying pH conditions.

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Main Results:

  • Permeabilized platelets generated 10-15 times more lipoxins than peptidoleukotrienes from LTA4, whereas intact platelets produced similar amounts.
  • Lipoxin A4 (LXA4), lipoxin B4 (LXB4), and their all-trans isomers were identified; chiral analysis showed specific R configurations.
  • Permeabilized platelets exhibited higher lipoxin generation (3-5 times) than intact cells, with optimal activity at pH 7-9. LXB4 formation was inhibited by LTA4, suggesting suicide inactivation.

Conclusions:

  • Lipoxin formation is a significant metabolic pathway for LTA4 in activated human platelets, particularly when membrane integrity is lost.
  • Platelet lipoxin generation is more efficient in permeabilized cells compared to intact cells.
  • Human endothelial cells do not convert LTA4 to lipoxins, highlighting the specificity of platelets in this pathway.