Naccache P H, Bourgoin S, Plante E, Roberge C J, de Medicis R, Lussier A, Poubelle P E
Centre de Recherche en Inflammation, Immunologie et Rhumatologie, CHUL, Ste.-Foy, Quebec, Canada.
Arthritis Rheum. 1993 Jan;36(1):117-25. doi: 10.1002/art.1780360119.
To investigate the involvement of phospholipase D in the signaling pathways activated by 2 pathologically relevant inflammatory microcrystals, monosodium urate (MSU) and calcium pyrophosphate dihydrate (CPPD).
Human peripheral blood neutrophils were used throughout. Phospholipase D activity was monitored by measuring 3 separate indices: 1) the mass of phosphatidic acid, 2) the levels of alkyl-phosphatidic acid, and 3) the levels of formation, in the presence of ethanol, of phosphatidylethanol. The latter 2 parameters were measured in cells labeled with 1-0-3H-alkyl-2-acetyl-sn-glycero-3-phosphocholine. The cells were stimulated with microcrystals of triclinic morphology.
Both MSU and CPPD crystals induced a time- and concentration-dependent accumulation of phosphatidic acid mass and elevation in levels of alkyl-phosphatidic acid and phosphatidylethanol in prelabeled cells. The activation of phospholipase D by the microcrystals was partially sensitive to colchicine and largely resistant to pertussis toxin. Inhibition of phosphatidic acid formation by wortmannin or ethanol reduced the microcrystal-stimulated production of superoxide anions.
These results indicate that microcrystals stimulate phospholipase D in human neutrophils and that at least some of the functional consequences of neutrophil-microcrystal interactions may be dependent on this biochemical pathway.
研究磷脂酶D在由两种病理相关炎症微晶——尿酸钠（MSU）和二水焦磷酸钙（CPPD）激活的信号通路中的作用。
全程使用人外周血中性粒细胞。通过测量3个独立指标监测磷脂酶D活性：1）磷脂酸的质量；2）烷基磷脂酸的水平；3）在乙醇存在下磷脂酰乙醇的生成水平。后两个参数在标记有1-0-³H-烷基-2-乙酰基-sn-甘油-3-磷酸胆碱的细胞中测量。用三斜晶形微晶刺激细胞。
MSU和CPPD晶体均诱导预标记细胞中磷脂酸质量随时间和浓度依赖性积累，以及烷基磷脂酸和磷脂酰乙醇水平升高。微晶对磷脂酶D的激活对秋水仙碱部分敏感，对百日咳毒素基本耐受。渥曼青霉素或乙醇对磷脂酸形成的抑制降低了微晶刺激的超氧阴离子产生。
这些结果表明微晶刺激人中性粒细胞中的磷脂酶D，并且中性粒细胞与微晶相互作用的至少一些功能后果可能依赖于这一生化途径。
