Vocks Andreas, Petković Marijana, Arnhold Jürgen
Institute of Medical Physics and Biophysics, Medical Faculty, University of Leipzig.
Cell Physiol Biochem. 2003;13(3):165-72. doi: 10.1159/000071867.
BACKGROUND/AIM: Polymorphonuclear leukocytes (neutrophils) release a variety of toxic agents--proteins and reactive oxygen species (ROS)--that are used to inactivate foreign microorganisms in the non-specific immune response. This study was undertaken to compare intracellular signalling pathways that lead to the ROS production as well as degranulation of azurophilic granules of human fMet-Leu-Phe/cytochalasin B stimulated neutrophils.
Luminol-amplified chemiluminescence was used for monitoring the oxidative activity of human neutrophils in the presence of various inhibitors. The elastase activity was assessed in the neutrophil supernatant as a marker for degranulation of azurophilic granules.
Tested inhibitors of enzymes of signalling cascades showed the same effect on the ROS production and on the activity of elastase released from neutrophils. The only difference was obtained with staurosporine: it inhibited the chemiluminescence response, but increased the elastase release.
Early signalling pathways leading to the ROS production and the degranulation are ubiquitous in human neutrophils. They are branching most probably at the point of the phosphatidic acid production by phospholipase D. A protein kinase activated by this lipid second messenger might play a central regulatory role in human neutrophils.
背景/目的：多形核白细胞（中性粒细胞）会释放多种毒性因子——蛋白质和活性氧（ROS），这些因子在非特异性免疫反应中用于使外来微生物失活。本研究旨在比较导致人fMet-Leu-Phe/细胞松弛素B刺激的中性粒细胞产生ROS以及嗜天青颗粒脱颗粒的细胞内信号通路。
鲁米诺增强化学发光法用于在存在各种抑制剂的情况下监测人中性粒细胞的氧化活性。中性粒细胞上清液中的弹性蛋白酶活性作为嗜天青颗粒脱颗粒的标志物进行评估。
测试的信号级联酶抑制剂对ROS产生和从中性粒细胞释放的弹性蛋白酶活性显示出相同的作用。唯一的差异是与星形孢菌素有关：它抑制化学发光反应，但增加弹性蛋白酶释放。
导致ROS产生和脱颗粒的早期信号通路在人中性粒细胞中普遍存在。它们很可能在磷脂酶D产生磷脂酸的点处分支。由这种脂质第二信使激活的蛋白激酶可能在人中性粒细胞中起核心调节作用。
