title: Increased lysophosphatidylcholine and non-esterified fatty acid content in LDL induces chemokine release in endothelial cells. Relationship with electronegative LDL.
abstract: Electronegative low-density lipoprotein (LDL(-)) is a plasma-circulating LDL subfraction with proinflammatory properties that induces the production of chemokines in cultured endothelial cells. However, the specific mechanism of LDL(-)-mediated chemokine release is presently unknown. A characteristic feature of LDL(-) is an increased content of lysophosphatidylcholine (LPC) and non-esterified fatty acids (NEFA). The effect of increasing amounts of LPC and NEFA associated with LDL on the release of chemokines by endothelial cells was studied. Total LDL was subfractionated by anion-exchange chromatography in electropositive (LDL(+)) and LDL(-). LDL(-) contained two-fold more LPC and NEFA than LDL(+) and induced two- to four-fold more (p < 0.05) interleukin-8 (IL-8, 11.5 +/- 8.2 ng/10(5) cells) and monocyte chemotactic protein-1 (MCP-1, 10.8 +/- 3.8 ng/10(5) cells) release by human umbilical vein endothelial cells (HUVEC) than LDL(+) (IL-8: 3.4 +/- 1.5 ng/10(5) cells, MCP-1: 5.8 +/- 2.9 ng/10(5) cells). The content of LPC and NEFA in LDL(+) was increased by enzymatic treatment with secretory phospholipase A(2) (sPLA(2)) at 5 ng/mL or 20 ng/mL or by incubation with NEFA at 2 mmol/L. Modification of LDL(+) by both methods did not result in oxidative modification as demonstrated by the lack of change in antioxidants, conjugated dienes and malondialdehyde content. sPLA(2) treatment resulted in an increase in LPC and NEFA in LDL(+) which enhanced its ability to release IL-8 and MCP-1 by HUVEC in a concentration-dependent manner (sPLA(2)(5)-LDL; IL-8: 7.1 +/- 3.8ng/10(5) cells, MCP-1: 8.0 +/- 5.1 ng/10(5) cells; sPLA(2)(20)-LDL; IL-8: 20.8 +/- 11.2 ng/10(5) cells, MCP-1: 15.0 +/- 7.5 ng/10(5) cells). NEFA loading of LDL(+) also favored the release of IL-8 and MCP-1 (IL-8: 7.8 +/- 6.1 ng/10(5) cells, MCP-1: 8.4 +/- 2.7 ng/10(5) cells, p < 0.05 versus LDL(+)). These effects were observed when modified LDL(+) reached a content of LPC and/or NEFA similar that of LDL(-). These data indicate that non-oxidized polar lipids associated with LDL promote an inflammatory response in endothelial cells and suggest that increased NEFA and LPC could be involved in the inflammatory activity of LDL(-).
authors.affiliation: Department of Biochemistry and Inflammation Mediators Laboratory, Institut de Recerca, Hospital de la Santa Creu i Sant Pau, C/ Antoni Maria Claret 167, Barcelona 08025, Spain.
authors.forename: Sònia
authors.initials: S
authors.lastname: Benítez
authors.forename: Mercedes
authors.initials: M
authors.lastname: Camacho
authors.forename: Rosa
authors.initials: R
authors.lastname: Arcelus
authors.forename: Luís
authors.initials: L
authors.lastname: Vila
authors.forename: Cristina
authors.initials: C
authors.lastname: Bancells
authors.forename: Jordi
authors.initials: J
authors.lastname: Ordóñez-Llanos
authors.forename: José Luis
authors.initials: JL
authors.lastname: Sánchez-Quesada
mesh_terms: D018932:Chemokine CCL2 / Q000378:metabolism; D018925:Chemokines / Q000378:metabolism*; D002852:Chromatography, Ion Exchange; D042783:Endothelial Cells / Q000378:metabolism*; D005230:Fatty Acids, Nonesterified / Q000097:blood*; D006801:Humans; D016209:Interleukin-8 / Q000378:metabolism; D008077:Lipoproteins, LDL / Q000097:blood*; D008244:Lysophosphatidylcholines / Q000097:blood*
chemical_list: C111835:CCL2 protein, human; D018932:Chemokine CCL2; D018925:Chemokines; D005230:Fatty Acids, Nonesterified; D016209:Interleukin-8; D008077:Lipoproteins, LDL; D008244:Lysophosphatidylcholines