Cominacini L, Rigoni A, Pasini A F, Garbin U, Davoli A, Campagnola M, Pastorino A M, Lo Cascio V, Sawamura T
Department of Biomedical and Surgical Sciences, Verona University, 37134 Verona, Italy.
J Biol Chem. 2001 Apr 27;276(17):13750-5. doi: 10.1074/jbc.M010612200. Epub 2001 Jan 24.
Oxidized low density lipoprotein (ox-LDL) has been suggested to affect endothelium-dependent vascular tone through a decreased biological activity of endothelium-derived nitric oxide (NO). Oxidative inactivation of NO is regarded as an important cause of its decreased biological activity, and in this context superoxide (O(2)) is known to inactivate NO in a chemical reaction during which peroxynitrite is formed. In this study we examined the effect of ox-LDL on the intracellular NO concentration in bovine aortic endothelial cells and whether this effect is influenced by ox-LDL binding to the endothelial receptor lectin-like ox-LDL receptor-1 (LOX-1) through the formation of reactive oxygen species and in particular of O(2). ox-LDL induced a significant dose-dependent decrease in intracellular NO concentration both in basal and stimulated conditions after less than 1 min of incubation with bovine aortic endothelial cells (p < 0.01). In the same experimental conditions ox-LDL also induced O(2) generation (p < 0.001). In the presence of radical scavengers and anti-LOX-1 monoclonal antibody, O(2) formation induced by ox-LDL was reduced (p < 0.001) with a contemporary rise in intracellular NO concentration (p < 0.001). ox-LDL did not significantly modify the ability of endothelial nitric oxide synthase to metabolize l-arginine to l-citrulline. The results of this study show that one of the pathophysiological consequences of ox-LDL binding to LOX-1 may be the inactivation of NO through an increased cellular production of O(2).
氧化型低密度脂蛋白(ox-LDL)被认为可通过降低内皮源性一氧化氮(NO)的生物活性来影响内皮依赖性血管张力。NO的氧化失活被视为其生物活性降低的一个重要原因,在这种情况下,超氧阴离子(O₂)已知在化学反应中使NO失活,在此过程中会形成过氧亚硝酸盐。在本研究中,我们检测了ox-LDL对牛主动脉内皮细胞内NO浓度的影响,以及这种影响是否受ox-LDL通过活性氧物质尤其是O₂的形成与内皮受体凝集素样ox-LDL受体-1(LOX-1)结合的影响。与牛主动脉内皮细胞孵育不到1分钟后,ox-LDL在基础和刺激条件下均诱导细胞内NO浓度出现显著的剂量依赖性降低(p < 0.01)。在相同实验条件下,ox-LDL还诱导了O₂的产生(p < 0.001)。在存在自由基清除剂和抗LOX-1单克隆抗体的情况下,ox-LDL诱导的O₂形成减少(p < 0.001),同时细胞内NO浓度升高(p < 0.001)。ox-LDL并未显著改变内皮型一氧化氮合酶将L-精氨酸代谢为L-瓜氨酸的能力。本研究结果表明,ox-LDL与LOX-1结合的病理生理后果之一可能是通过细胞O₂产生增加使NO失活。