Coleman Leon G, Polanowska-Grabowska Renata K, Marcinkiewicz Marek, Gear Adrian R L
Department of Biochemistry and Molecular Genetics, University of Virginia Health Sciences Center, Charlottesville, VA 22908, USA.
Blood. 2004 Jul 15;104(2):380-9. doi: 10.1182/blood-2003-08-2961. Epub 2004 Mar 30.
The in vitro oxidation of low-density lipoprotein (LDL) by hypochlorous acid produces a modified form (HOCl-LDL) capable of stimulating platelet function. We now report that HOCl-LDL is highly effective at inducing platelet function, causing stable aggregation and alpha-granule secretion. Such stimulation depended on the presence of low levels of primary agonists such as adenosine diphosphate (ADP) and thrombin, or others like epinephrine (EPI) and macrophage-derived chemokine (MDC, CCL22). Agonist levels, which by themselves induced little or reversible aggregation, caused strong stable aggregation when combined with low levels of HOCl-LDL. Platelet activation by HOCl-LDL and ADP (1 microM) caused P-selectin (CD62P) exposure, without serotonin or adenosine triphosphate (ATP) secretion. Intracellular calcium levels rose slowly (from 100 to 200 nM) in response to HOCl-LDL alone and rapidly when combined with ADP to about 300 nM. p38 mitogen-activated protein kinase (MAPK) became phosphorylated in response to HOCl-LDL alone. This phosphorylation was not blocked by the protein kinase C (PKC) inhibitor bisindolylmaleimide, which reduced the extent of aggregation and calcium increase. However, the p38 MAPK inhibitor SB203580 blocked platelet aggregation and phosphorylation of p38 MAPK. These findings suggest that HOCl-LDL exposed during atherosclerotic plaque rupture, coupled with low levels of primary agonists, can rapidly induce extensive and stable thrombus formation.
次氯酸对低密度脂蛋白(LDL)进行体外氧化会产生一种能够刺激血小板功能的修饰形式(HOCl-LDL)。我们现在报告,HOCl-LDL在诱导血小板功能方面非常有效,可导致稳定的聚集和α-颗粒分泌。这种刺激依赖于低水平的主要激动剂,如二磷酸腺苷(ADP)和凝血酶,或其他如肾上腺素(EPI)和巨噬细胞衍生趋化因子(MDC,CCL22)的存在。单独使用时只能诱导少量或可逆聚集的激动剂水平,与低水平的HOCl-LDL联合使用时会导致强烈的稳定聚集。HOCl-LDL和ADP(1 microM)激活血小板会导致P-选择素(CD62P)暴露,但不会分泌5-羟色胺或三磷酸腺苷(ATP)。单独使用HOCl-LDL时细胞内钙水平缓慢升高(从100 nM升至200 nM),与ADP联合使用时则迅速升高至约300 nM。单独使用HOCl-LDL时p38丝裂原活化蛋白激酶(MAPK)会发生磷酸化。这种磷酸化不会被蛋白激酶C(PKC)抑制剂双吲哚马来酰胺阻断,后者会降低聚集程度和钙升高幅度。然而,p38 MAPK抑制剂SB203580可阻断血小板聚集和p38 MAPK的磷酸化。这些发现表明,动脉粥样硬化斑块破裂时暴露的HOCl-LDL,与低水平的主要激动剂相结合,可迅速诱导广泛而稳定的血栓形成。