Gugiu B Gabriel, Mouillesseaux Kevin, Duong Victoria, Herzog Tabitha, Hekimian Avetis, Koroniak Lukasz, Vondriska Thomas M, Watson Andrew D
Department of Pathology and Laboratory Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA.
J Lipid Res. 2008 Mar;49(3):510-20. doi: 10.1194/jlr.M700264-JLR200. Epub 2007 Dec 10.
Oxidation products of 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphatidylcholine (Ox-PAPC) are found in atherosclerotic lesions, apoptotic cells, and oxidized LDL and stimulate human aortic endothelial cells (HAECs) to produce inflammatory cytokines, leukocyte chemoattractants, and coagulation factors. This regulation is thought to be a receptor-mediated process in which oxidized phospholipids activate specific receptors on HAECs to evoke an inflammatory response. To characterize the HAEC proteins with which oxidized phospholipids interact, a biotinylated PAPC analog, 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphatidyl-(N-biotinylethanolamine) (PAPE-N-biotin), was synthesized. Oxidation of PAPE-N-biotin in air generated a mixture of biotin-labeled oxidized lipids analogous to Ox-PAPC. Ox-PAPE-N-biotin, like Ox-PAPC, induced interleukin-8 (IL-8) protein synthesis and stimulated IL-8, low density lipoprotein receptor, heme oxygenase-1, and activating transcription factor-3 mRNA expression in HAECs. After treatment of HAECs with Ox-PAPE-N-biotin, the cellular proteins were isolated and separated by SDS-PAGE. Western analysis with streptavidin-HRP demonstrated at least 20 different biotinylated HAEC proteins to which the Ox-PAPE-N-biotin was associated, which were not detected with unoxidized PAPE-N-biotin treatment. This work suggests that oxidized phospholipids, such as those found in oxidized LDL, apoptotic cells, and atherosclerotic lesions, form tight interactions with specific endothelial cell proteins, which may be responsible for the inflammatory response. Identification of these putative oxidized phospholipid targets may reveal therapeutic targets to modulate inflammation and atherosclerosis.
1-棕榈酰-2-花生四烯酰-sn-甘油-3-磷脂酰胆碱（Ox-PAPC）的氧化产物存在于动脉粥样硬化病变、凋亡细胞和氧化型低密度脂蛋白中，并刺激人主动脉内皮细胞（HAECs）产生炎性细胞因子、白细胞趋化因子和凝血因子。这种调节被认为是一个受体介导的过程，其中氧化磷脂激活HAECs上的特定受体以引发炎症反应。为了鉴定与氧化磷脂相互作用的HAEC蛋白，合成了一种生物素化的PAPC类似物，即1-棕榈酰-2-花生四烯酰-sn-甘油-3-磷脂酰-（N-生物素乙醇胺）（PAPE-N-生物素）。PAPE-N-生物素在空气中氧化产生了一种生物素标记的氧化脂质混合物，类似于Ox-PAPC。Ox-PAPE-N-生物素与Ox-PAPC一样，诱导白细胞介素-8（IL-8）蛋白合成，并刺激HAECs中IL-8、低密度脂蛋白受体、血红素加氧酶-1和激活转录因子-3的mRNA表达。用Ox-PAPE-N-生物素处理HAECs后，分离细胞蛋白并通过SDS-PAGE进行分离。用链霉亲和素-HRP进行的Western分析表明，至少有20种不同的生物素化HAEC蛋白与Ox-PAPE-N-生物素相关，而未氧化的PAPE-N-生物素处理未检测到这些蛋白。这项工作表明，氧化磷脂，如在氧化型低密度脂蛋白、凋亡细胞和动脉粥样硬化病变中发现的那些，与特定的内皮细胞蛋白形成紧密相互作用，这可能是炎症反应的原因。鉴定这些假定的氧化磷脂靶点可能揭示调节炎症和动脉粥样硬化的治疗靶点。
