Holland J A, Meyer J W, Chang M M, O'Donnell R W, Johnson D K, Ziegler L M
Department of Medicine, State University of New York Health Science Center, Syracuse 13210, USA.
Endothelium. 1998;6(2):113-21. doi: 10.3109/10623329809072198.
In order to study the major cellular source of reactive oxygen species (ROS) in perturbed human endothelial cells (EC), the effect of thrombin, a phospholipase A2 activator, on cultured EC ROS generation has been investigated. EC were incubated with 0.1-1 unit/ml thrombin and cellular superoxide anion (O(-)2) release and hydrogen peroxide (H2O2) production measured. Thrombin exposure caused an elevation in EC O(-)2 release and H2O2 production. The effects of protein kinase C, arachidonic acid metabolism, NADPH oxidase, and phospholipase A2 inhibitors on thrombin-induced EC H2O2 production were examined. EC were exposed to 0.5 unit/ml thrombin and cellular H2O2 production measured in the presence and absence of the protein kinase C inhibitor, H-7; arachidonic acid metabolism inhibitors, indomethacin, nordihydroguaiaretic acid, and SKF525A; NADPH oxidase inhibitor, apocynin; and phospholipase A2 inhibitor, 4-bromophenacyl bromide. All inhibitors, with the exception of H-7 and indomethacin, suppressed thrombin-induced EC H2O2 production. The pattern of effects of these metabolic antagonists on thrombin-induced EC ROS production is similar to that previously reported on ROS production in EC exposed to high low-density lipoprotein levels, and in stimulated leukocytes. These findings further implicate NADPH oxidase as a major ROS source in EC.
为了研究人内皮细胞（EC）受到干扰时活性氧（ROS）的主要细胞来源，我们研究了凝血酶（一种磷脂酶A2激活剂）对培养的EC产生ROS的影响。将EC与0.1 - 1单位/毫升的凝血酶孵育，并测量细胞超氧阴离子（O(-)2）释放和过氧化氢（H2O2）生成。凝血酶暴露导致EC的O(-)2释放和H2O2生成增加。我们检测了蛋白激酶C、花生四烯酸代谢、NADPH氧化酶和磷脂酶A2抑制剂对凝血酶诱导的EC产生H2O2的影响。将EC暴露于0.5单位/毫升的凝血酶，并在存在和不存在蛋白激酶C抑制剂H - 7、花生四烯酸代谢抑制剂吲哚美辛、去甲二氢愈创木酸和SKF525A、NADPH氧化酶抑制剂鱼藤酮以及磷脂酶A2抑制剂4 - 溴苯甲酰溴的情况下测量细胞H2O2生成。除H - 7和吲哚美辛外，所有抑制剂均抑制凝血酶诱导的EC产生H2O2。这些代谢拮抗剂对凝血酶诱导的EC产生ROS的影响模式与先前报道的EC暴露于高低密度脂蛋白水平以及刺激的白细胞中ROS产生的影响模式相似。这些发现进一步表明NADPH氧化酶是EC中ROS的主要来源。
