Carluccio M A, Massaro M, Bonfrate C, Siculella L, Maffia M, Nicolardi G, Distante A, Storelli C
CNR Institute of Clinical Physiology, Lecce, Italy.
Arterioscler Thromb Vasc Biol. 1999 Feb;19(2):220-8. doi: 10.1161/01.atv.19.2.220.
Because oleic acid is implicated in the antiatherogenic effects attributed to the Mediterranean diet, we investigated whether this fatty acid can modulate endothelial activation, ie, the concerted expression of gene products involved in leukocyte recruitment and early atherogenesis. We incubated sodium oleate with human umbilical vein endothelial cells for 0 to 72 hours, followed by coincubation of oleate with human recombinant tumor necrosis factor, interleukin (IL)-1alpha, IL-1beta, IL-4, Escherichia coli lipopolysaccharide (LPS), or phorbol 12-myristate 13-acetate for a further 6 to 24 hours. The endothelial expression of vascular cell adhesion molecule-1 (VCAM-1), E-selectin, and intercellular adhesion molecule-1 was monitored by cell surface enzyme immunoassays or flow cytometry, and steady-state levels of VCAM-1 mRNA were assessed by Northern blot analysis. At 10 to 100 micromol/L for >24 hours, oleate inhibited the expression of all adhesion molecules tested. After a 72-hour incubation with oleate and a further 16-hour incubation with oleate plus 1 microg/mL LPS, VCAM-1 expression was reduced by >40% compared with control. Adhesion of monocytoid U937 cells to LPS-treated endothelial cells was reduced concomitantly. Oleate also produced a quantitatively similar reduction of VCAM-1 mRNA levels on Northern blot analysis and inhibited nuclear factor-kappaB activation on electrophoretic mobility shift assays. Incubation of endothelial cells with oleate for 72 hours decreased the relative proportions of saturated (palmitic and stearic) acids in total cell lipids and increased the proportions of oleate in total cell lipids without significantly changing the relative proportions of polyunsaturated fatty acids. Although less potent than polyunsaturated fatty acids in inhibiting endothelial activation, oleic acid may contribute to the prevention of atherogenesis through selective displacement of saturated fatty acids in cell membrane phospholipids and a consequent modulation of gene expression for molecules involved in monocyte recruitment.
由于油酸与地中海饮食的抗动脉粥样硬化作用有关，我们研究了这种脂肪酸是否能调节内皮细胞活化，即参与白细胞募集和早期动脉粥样硬化的基因产物的协同表达。我们将油酸钠与人脐静脉内皮细胞孵育0至72小时，然后将油酸与人重组肿瘤坏死因子、白细胞介素（IL）-1α、IL-1β、IL-4、大肠杆菌脂多糖（LPS）或佛波醇12-肉豆蔻酸酯13-乙酸酯再共同孵育6至24小时。通过细胞表面酶免疫测定或流式细胞术监测血管细胞黏附分子-1（VCAM-1）、E-选择素和细胞间黏附分子-1的内皮表达，并通过Northern印迹分析评估VCAM-1 mRNA的稳态水平。在10至100 μmol/L浓度下处理超过24小时，油酸可抑制所有测试黏附分子的表达。在用油酸孵育72小时并再用油酸加1 μg/mL LPS孵育16小时后，与对照相比，VCAM-1表达降低了40%以上。单核细胞样U937细胞与LPS处理的内皮细胞的黏附也随之减少。Northern印迹分析显示，油酸还使VCAM-1 mRNA水平出现了类似程度的降低，并在电泳迁移率变动分析中抑制了核因子-κB的活化。用油酸将内皮细胞孵育72小时可降低总细胞脂质中饱和（棕榈酸和硬脂酸）酸的相对比例，并增加总细胞脂质中油酸的比例，而多不饱和脂肪酸的相对比例没有明显变化。尽管在抑制内皮细胞活化方面不如多不饱和脂肪酸有效，但油酸可能通过选择性取代细胞膜磷脂中的饱和脂肪酸以及随后调节参与单核细胞募集的分子的基因表达，从而有助于预防动脉粥样硬化。
