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Title:Effect of Fatty Acid Modification on Prostacyclin Production in Cultured Human Endothelial Cells (Arachidonate, Elcosapentaenoate)
Author(s):Basgall, Edward Joseph
Department / Program:Food Science
Discipline:Food Science
Degree Granting Institution:University of Illinois at Urbana-Champaign
Degree:Ph.D.
Genre:Dissertation
Subject(s):Health Sciences, Nutrition
Abstract:An endothelial cell culture system was used to compare the effects of endogenous arachidonic acid (AA) and eicosapentaenoic acid (EPA) on prostacyclin synthesis. Supplemental levels of fatty acids (20-150 (mu)M) were supplied as sodium salts bound to bovine serum albumin (3:1 mole ratio) and added to M-199 with 20% fetal bovine serum. Both AA and EPA were readily incorporated into neutral and phospholipid fractions to serve as endogenous precursors for prostacyclin (PGI(,2)) synthesis. Thrombin (.5 U/ml) was used to stimulate prostacyclin synthesis. AA enrichment did not result in an increased PGI(,2) synthesis while EPA enrichment decreased PGI(,2) synthesis as measured by radioimmunoassay to 6-keto-PGF(,1(alpha)). Radiolabelled fatty acid eicosanoid products were separated by reverse phase HPLC. Unconverted fatty acid was the major product released upon thrombin stimulation in both cases. ('3)H-AA supplemented cells also released PGI(,2) and HETE in lesser amounts while ('14)C-EPA supplemented cells released only minimal amounts of PGI(,3) and HEPE. The relative percent incorporation of radiolabelled ('3)H-AA and ('14)C-EPA into cellular lipids was determined after two-dimensional thin-layer chromatography. Neutral lipid contained the vast majority of radiolabel ((TURN)50%). Of the phospholipids, phosphatidylcholine contained the greatest percentage of radiolabel. Phosphatidylinositol and phosphatidylethanolamine contained intermediate percentages while phosphatidylserine and sphingomyelin contained the least. In a dual label supplementation regimen (150 (mu)M ('14)C-EPA and 16 (mu)M ('3)H-AA), phosphatidylinositol appeared to be readily saturated with fatty acid at an early stage. This may be an important regulatory aspect for prostaglandin synthesis.
Issue Date:1983
Type:Text
Language:English
Description:153 p.
Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 1983.
URI:http://hdl.handle.net/2142/77444
Other Identifier(s):(UMI)AAI8409859
Date Available in IDEALS:2015-05-13
Date Deposited:1983


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