Synthesis of 1-acyl-2-(3H)acetyl-SN-glycero-3-phosphocholine, a structural analog of platelet activating factor, by vascular endothelial cells

Journal Article · Wed May 15 04:00:00 EDT 1991 · Biochemical and Biophysical Research Communications; (United States)

DOI:https://doi.org/10.1016/0006-291X(91)90465-J· OSTI ID:5596008

Mueller, H W; Nollert, M U; Eskin, S G ^[1]

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1. Division of Cardiology, University of Texas Health Science Center, Houston (USA)

Human umbilical vein endothelial cells (HUVECS) were challenged with thrombin in the presence of (3H)acetate to stimulate the production of radiolabeled platelet activating factor (PAF, 1-O-alkyl-2-(3H)acetyl-sn-glycero-3-phosphocholine, 1-O-alkyl-2-(3H)acetyl-GPC). The 3H-product was isolated by thin-layer chromatography, and 1-radyl-2(3H),3- diacetylglycerols were prepared by phospholipase C digestion and subsequent acetylation at the sn-3 position. When the 1-radyl-2(3H),3-diacetylglycerols were analyzed by zonal thin-layer chromatography, 96-97% of the radiolabeled derivative migrated with 1-acyl-2,3-diacetylglycerol standard. Only minor amounts (3-4%) of 1-alkyl-2(3H),3-diacetylglycerol were observed, demonstrating that the predominant acetylated product synthesized by thrombin-stimulated HUVECS was 1-acyl-2-(3H)acetyl-GPC. This relative abundance of 1-acyl-2-(3H)-acetyl-GPC was not significantly affected by thrombin dose, incubation time, or cell passage, and was also observed in HUVECS challenged with ionophore A23187. In addition, the acetylated product from ionophore A23187- or bradykinin-stimulated bovine aortic endothelial cells contained 90% 1-acyl-2-(3H)acetyl-GPC, suggesting that the synthesis of the 1-acyl PAF analog is not unique to HUVECS. These findings demonstrate that PAF is a minor synthetic component of HUVECS and bovine aortic endothelial cells. In light of the integral role which the vascular endothelial cell plays in the regulation of thrombosis, these findings also suggest that the production of 1-acyl-2-acetyl-GPC may be biologically important.

• OSTI ID: 5596008
• Journal Information: Biochemical and Biophysical Research Communications; (United States), Journal Name: Biochemical and Biophysical Research Communications; (United States) Vol. 176:3; ISSN BBRCA; ISSN 0006-291X
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ACETATES
ACETYLATION
ACYLATION
ANIMAL TISSUES
ANIMALS
AORTA
ARTERIES
BIOSYNTHESIS
BLOOD VESSELS
BODY
CARBOXYLIC ACID SALTS
CARDIOVASCULAR DISEASES
CARDIOVASCULAR SYSTEM
CATTLE
CELL CULTURES
CHEMICAL REACTIONS
CHROMATOGRAPHY
DISEASES
DOMESTIC ANIMALS
ENDOTHELIUM
ESTERS
HYDROGEN COMPOUNDS
ISOTOPE APPLICATIONS
LIPIDS
MAMMALS
MAN
ORGANIC COMPOUNDS
ORGANIC PHOSPHORUS COMPOUNDS
ORGANS
PATHOGENESIS
PHOSPHOLIPIDS
PRIMATES
RUMINANTS
SEPARATION PROCESSES
SYNTHESIS
THIN-LAYER CHROMATOGRAPHY
THROMBOSIS
TISSUES
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VASCULAR DISEASES
VEINS
VERTEBRATES