Heemskerk J W, Feijge M A, Kalafusz R, Hornstra G
Department of Biochemistry, University of Limburg, Maastricht, The Netherlands.
Biochim Biophys Acta. 1989 Aug 8;1004(2):252-60. doi: 10.1016/0005-2760(89)90275-0.
The apparent steady-state fluorescence anisotropy of DPH- or TMA-DPH-labeled washed rat platelets is strongly affected by factors that also influence the turbidity by these platelet suspensions. Sonicated preparations from platelet lipids have a low turbidity and give anisotropy values which are hardly affected by the experimental conditions. We studied the effect of four high-fat diets on membrane fluidity, lipid composition and activation tendency of washed platelets. The diets contained 50 energy% of oils with different levels of saturated and (poly)unsaturated fatty acids. Only small diet-induced differences in DPH fluorescence anisotropy were found, which were comparable for intact platelets and platelet lipids. These differences were unrelated to the degree of saturation of the dietary fatty acids. Platelets from rats fed mainly saturated fatty acids differed significantly from other diet groups in a higher unsaturation degree of phospholipids and a lower cholesterol/phospholipid ratio, but this was not detected by DPH in terms of decreased anisotropy. These platelets aggregated less than other platelets in response to thrombin or collagen. The lower response to collagen persisted in indomethacin-treated platelets activated with the thromboxane A2 mimetic U46619, indicating a different sensitivity of these platelets for thromboxane A2. We conclude that in rat platelets: (a) the overall membrane fluidity and phospholipid unsaturation degree are subject to strong homeostatic control; (b) steady-state anisotropy with DPH or TMA-DPH label is inadequate to reveal subtile changes in lipid profile; (c) changes in platelet responsiveness to thrombin and thromboxane A2, rather than (plasma) membrane fluidity, determine the effect of dietary fatty acids on platelet aggregation.
用DPH或TMA - DPH标记的洗涤大鼠血小板的表观稳态荧光各向异性受那些也会影响这些血小板悬液浊度的因素的强烈影响。血小板脂质的超声处理制剂浊度低,其给出的各向异性值几乎不受实验条件的影响。我们研究了四种高脂饮食对洗涤血小板的膜流动性、脂质组成和活化倾向的影响。这些饮食含有50%能量的不同饱和及(多)不饱和脂肪酸水平的油。仅发现饮食诱导的DPH荧光各向异性存在微小差异,完整血小板和血小板脂质的差异相当。这些差异与饮食脂肪酸的饱和度无关。主要饲喂饱和脂肪酸的大鼠的血小板与其他饮食组相比,磷脂的不饱和度更高,胆固醇/磷脂比更低,但用DPH未检测到各向异性降低。这些血小板对凝血酶或胶原的聚集反应比其他血小板少。在用血栓素A2模拟物U46619激活的吲哚美辛处理的血小板中,对胶原的较低反应仍然存在,表明这些血小板对血栓素A2的敏感性不同。我们得出结论,在大鼠血小板中:(a)整体膜流动性和磷脂不饱和度受到强大的稳态控制;(b)用DPH或TMA - DPH标记的稳态各向异性不足以揭示脂质谱的细微变化;(c)血小板对凝血酶和血栓素A2反应性的变化,而非(血浆)膜流动性,决定了饮食脂肪酸对血小板聚集的影响。