title: Role of caveolin-1 in EGCG-mediated protection against linoleic-acid-induced endothelial cell activation.
abstract: Flavonoids can protect against inflammatory diseases such as atherosclerosis by decreasing vascular endothelial cell activation. Plasma microdomains called caveolae may be critical in regulating endothelial activation. Caveolae are particularly abundant in endothelial cells and play a major role in endothelial trafficking and the regulation of signaling pathways associated with the pathology of vascular diseases. We hypothesize that flavonoids can down-regulate endothelial inflammatory parameters by modulating caveolae-regulated cell signaling. We focused on the role of caveolae and its major protein, caveolin-1, in mechanisms of linoleic-acid-induced endothelial cell activation and protection by the catechin epigallocatechin-3-gallate (EGCG). Exposure to linoleic acid for 6 h induced expression of both caveolin-1 and cyclooxygenase (COX)-2. Pretreatment with EGCG blocked fatty-acid-induced caveolin-1 and COX-2 expression in a time- and concentration-dependent manner. Similar results were observed with nuclear factor-kappa B DNA binding activity, which was also reduced by caveolin-1 silencing. Exposure to linoleic acid rapidly increased phosphorylation of several kinases, including p38 MAPK, extracellular signal regulated kinase 1/2 (ERK1/2) and amino kinase terminal (Akt), with maximal induction at about 10 min. Inhibitors of ERK1/2 and Akt down-regulated the linoleic-acid-induced increase in COX-2 protein, which also occurred after pretreatment with EGCG. Caveolin-1 silencing blocked linoleic-acid-induced phosphorylation of ERK1/2 and protein expression of COX-2, suggesting that specific MAPK signaling is caveolae dependent. Our data provide evidence that caveolae may play a critical role in regulating vascular endothelial cell activation and protection by flavonoids such as EGCG.
authors.affiliation: Graduate Center for Nutritional Sciences, University of Kentucky, Lexington, 40536-0200, USA.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Zheng
authors.forename: Eum Jin
authors.initials: EJ
authors.lastname: Lim
authors.forename: Lei
authors.initials: L
authors.lastname: Wang
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Smart
authors.forename: Michal
authors.initials: M
authors.lastname: Toborek
authors.forename: Bernhard
authors.initials: B
authors.lastname: Hennig
mesh_terms: D002392:Catechin / Q000031:analogs & derivatives* / Q000494:pharmacology; D051242:Caveolin 1 / Q000502:physiology*; D002460:Cell Line; D051546:Cyclooxygenase 2 / Q000096:biosynthesis; D015536:Down-Regulation; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000378:metabolism*; D006801:Humans; D019787:Linoleic Acid / Q000494:pharmacology*; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D016328:NF-kappa B / Q000187:drug effects / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D034741:RNA, Small Interfering / Q000494:pharmacology; D015854:Up-Regulation
chemical_list: D051242:Caveolin 1; D016328:NF-kappa B; D034741:RNA, Small Interfering; D002392:Catechin; D019787:Linoleic Acid; C045651:epigallocatechin gallate; D051546:Cyclooxygenase 2; D051057:Proto-Oncogene Proteins c-akt; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3