Kouoh F, Gressier B, Luyckx M, Brunet C, Dine T, Ballester L, Cazin M, Cazin J C
Laboratoire de Pharmacologie, Pharmacocinétique et Pharmacie Clinique, rue du Professeur Laguesse, BP 83, 59006 Lille, France.
Biol Pharm Bull. 2000 Nov;23(11):1382-3. doi: 10.1248/bpb.23.1382.
We report the successful application to human venous blood of a novel method developed to purify rabbit polymorphonuclear neutrophils (PMNs) from whole blood. Human PMNs were separated from whole blood after sedimentation with dextran and histopaque density gradient centrifugation. 3.92 +/- 0.26 x 10(6) PMNs per ml of blood was harvested. The purity of the preparation was 92.00 +/- 1.10%. The PMNs isolated were capable of generating a high amount of reactive oxygen species (ROS) and elastase after stimulation with phorbol 12-myristate 13-acetate (PMA): 13.43 +/- 0.3 microM of O2-, 9.62 +/- 0.15 microM of H2O2 and 5.48 +/- 0.01 microM of elastase. This method gives equivalent yield and viability when applied to isolating human or rabbit PMNs, in comparison with standard methods used to isolate human PMNs. Our method could be usefully exploited for comparative studies of rabbit and human PMNs with a cellular model of inflammatory oxidative stress in which the monitoring parameters are ROS and elastase. Thus, the results of animal (rabbit) studies can be extended to human diseases.
我们报告了一种从全血中纯化兔多形核中性粒细胞（PMN）的新方法在人静脉血中的成功应用。人PMN通过葡聚糖沉淀和Histopaque密度梯度离心从全血中分离出来。每毫升血液收获了3.92±0.26×10⁶个PMN。制备物的纯度为92.00±1.10%。分离出的PMN在用佛波酯12-肉豆蔻酸酯13-乙酸酯（PMA）刺激后能够产生大量活性氧（ROS）和弹性蛋白酶：超氧阴离子为13.43±0.3微摩尔，过氧化氢为9.62±0.15微摩尔，弹性蛋白酶为5.48±0.01微摩尔。与用于分离人PMN的标准方法相比，该方法在应用于分离人或兔PMN时具有相当的产量和活力。我们的方法可有效地用于兔和人PMN在炎症氧化应激细胞模型中的比较研究，其中监测参数为ROS和弹性蛋白酶。因此，动物（兔）研究的结果可以扩展到人类疾病。
