Görög P, Pearson J D, Kakkar V V
Thrombosis Research Unit, King's College School of Medicine and Dentistry, London, U.K.
Atherosclerosis. 1988 Jul;72(1):19-27. doi: 10.1016/0021-9150(88)90058-5.
We have studied the ability of particulate stimuli to induce the release of reactive oxygen metabolites from sub-cultured monolayers of human endothelial cells. Basal release of superoxide (O2-) and hydrogen peroxide from undisturbed monolayers was very low (108 pmol O2- and 75 pmol H2O2 in 3 h from dishes of 3 X 10(5) cells). Addition of 1-micron diameter polystyrene microspheres, which were phagocytosed by the cells progressively, caused a dramatic increase in release of both metabolites; by 3 h, a 13.5- and 6.6-fold increase over controls was observed respectively (P less than 0.001). Addition of formaldehyde-fixed human platelets or chylomicron-size lipid particles also increased production of reactive oxygen species. Similar rises in H2O2 and O2- production were induced by treatment with 10(-7) M phorbol myristate acetate. Pretreatment of endothelial cells with neuraminidase, heparinase or heparitinase to alter their glycocalyx composition substantially enhanced the effect of microspheres on H2O2 and O2- generation. We conclude that the interactions of particles, including platelets and lipids, with endothelial cells leads to the generation of significant pericellular levels of reactive oxygen species. These metabolites can oxidise a wide variety of nearby molecules, leading to cell damage and altered uptake characteristics for lipoproteins containing peroxidized lipids. These effects are exacerbated when endothelial cell glycocalyx composition is disrupted.
我们研究了颗粒性刺激物诱导人内皮细胞传代单层释放活性氧代谢产物的能力。未受干扰的单层细胞中超氧化物（O₂⁻）和过氧化氢的基础释放量非常低（在含有3×10⁵个细胞的培养皿中，3小时内分别为108皮摩尔O₂⁻和75皮摩尔H₂O₂）。添加直径为1微米的聚苯乙烯微球，细胞会逐渐吞噬这些微球，这导致两种代谢产物的释放显著增加；到3小时时，分别观察到比对照组增加了13.5倍和6.6倍（P＜0.001）。添加甲醛固定的人血小板或乳糜微粒大小的脂质颗粒也会增加活性氧的产生。用10⁻⁷M佛波醇肉豆蔻酸酯乙酸盐处理也会诱导H₂O₂和O₂⁻产生类似的增加。用神经氨酸酶、肝素酶或类肝素酶预处理内皮细胞以显著改变其糖萼组成，可大大增强微球对H₂O₂和O₂⁻生成的影响。我们得出结论，包括血小板和脂质在内的颗粒与内皮细胞的相互作用会导致细胞周围产生大量的活性氧。这些代谢产物可以氧化多种附近的分子，导致细胞损伤并改变对含有过氧化脂质的脂蛋白的摄取特性。当内皮细胞糖萼组成被破坏时，这些影响会加剧。
