Salvemini D, de Nucci G, Gryglewski R J, Vane J R
William Harvey Research Institute, Saint Bartholomew's Hospital Medical College, London, United Kingdom.
Proc Natl Acad Sci U S A. 1989 Aug;86(16):6328-32. doi: 10.1073/pnas.86.16.6328.
Incubation of neutrophils or mononuclear cells with washed platelets (all prepared from human venous blood) resulted in an inhibition of thrombin-induced platelet aggregation that was dependent on the number of nucleated cells added. The inhibition was potentiated by superoxide dismutase and reversed by oxyhemoglobin. In the case of neutrophils the inhibition was associated with an increase in cGMP, whereas with mononuclear cells both cAMP and cGMP were increased. The inhibitory activity of neutrophils or mononuclear cells was prevented by their preincubation with NG-monomethyl-L-arginine methyl ester. L-Arginine reversed the action of NG-monomethyl-L-arginine methyl ester, whereas D-arginine was ineffective. Preincubation of the cells with catalase or mannitol did not prevent their inhibitory action on platelet aggregation. The inhibition of platelet aggregation was not due to platelet damage or to uptake of thrombin by neutrophils or mononuclear cells. It was overcome by increasing the concentration of thrombin and was absent in cell-free supernatants obtained from a suspension of neutrophils or mononuclear cells or from mixtures of platelets with neutrophils or platelets with mononuclear cells. These data provide evidence for the release of a nitric oxide-like factor from human neutrophils and mononuclear cells. In addition, evidence is provided that, as in stimulated murine macrophages and endothelial cells, the precursor of this factor is L-arginine.
用洗涤过的血小板（均由人静脉血制备）与中性粒细胞或单核细胞共同孵育，会导致凝血酶诱导的血小板聚集受到抑制，这种抑制作用取决于所添加的有核细胞数量。超氧化物歧化酶可增强这种抑制作用，而氧合血红蛋白则可逆转这种抑制作用。就中性粒细胞而言，这种抑制作用与环鸟苷酸（cGMP）的增加有关，而对于单核细胞，环磷酸腺苷（cAMP）和cGMP均增加。中性粒细胞或单核细胞的抑制活性可通过其与NG-单甲基-L-精氨酸甲酯预孵育而被阻止。L-精氨酸可逆转NG-单甲基-L-精氨酸甲酯的作用，而D-精氨酸则无效。用过氧化氢酶或甘露醇对细胞进行预孵育并不能阻止它们对血小板聚集的抑制作用。血小板聚集的抑制并非由于血小板损伤或中性粒细胞或单核细胞对凝血酶的摄取。通过增加凝血酶浓度可克服这种抑制作用，并且在从中性粒细胞或单核细胞悬液或血小板与中性粒细胞或血小板与单核细胞的混合物中获得的无细胞上清液中不存在这种抑制作用。这些数据为人类中性粒细胞和单核细胞释放一氧化氮样因子提供了证据。此外，还提供了证据表明，与受刺激的小鼠巨噬细胞和内皮细胞一样，该因子的前体是L-精氨酸。
