Stahl G L, Morse D S, Martin S L
Department of Anesthesia, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.
Am J Physiol. 1997 Jun;272(6 Pt 1):C1821-8. doi: 10.1152/ajpcell.1997.272.6.C1821.
Polymorphonuclear neutrophils (PMN) and platelets interact to produce both inflammatory and anti-inflammatory lipid mediators during human disease. Because swine models of human disease are used, it is important to understand the mechanisms involved in the formation of lipid mediators from porcine PMN-platelet interactions. In the present study, we investigated the mechanism of thromboxane (Tx) A2 and lipoxin A4 (LXA4) formation from porcine PMN and platelets, respectively. PMN (10(7)/ml) and platelet (30 x 10(7)/ml) suspensions stimulated with porcine C5a (pC5a), but not recombinant human C5a (rhC5a), significantly enhanced TxB2 formation. After cytochalasin B treatment, pC5a or rhC5a significantly and equally enhanced TxB2 formation from PMN-platelet suspensions. A-23187-induced TxB2 formation from platelets was not significantly augmented by the presence of PMN in these suspensions. A-23187 induced significant LXA4 production from porcine PMN that was not augmented by addition of platelets. Flow cytometric analysis of PMN-platelet suspensions revealed activated platelets adherent to PMN following pC5a stimulation. CV-6209, a platelet-activating factor (PAF) receptor antagonist, dose dependently prevented pC5a-induced platelet adherence to PMN and TxB2 formation. These data demonstrate that 1) porcine PMN alone can biosynthesize LXA4 without the assistance of platelets, which is in sharp contrast to human PMN-platelet interactions, and 2) in the absence of cytochalasin B, pC5a stimulates PAF biosynthesis from porcine PMN, resulting in TxB2 formation from platelets.
在人类疾病过程中,多形核中性粒细胞(PMN)和血小板相互作用产生促炎和抗炎脂质介质。由于使用了人类疾病的猪模型,了解猪PMN-血小板相互作用中脂质介质形成的机制很重要。在本研究中,我们分别研究了猪PMN和血小板形成血栓素(Tx)A2和脂氧素A4(LXA4)的机制。用猪C5a(pC5a)而非重组人C5a(rhC5a)刺激PMN(10⁷/ml)和血小板(30×10⁷/ml)悬液,可显著增强TxB2的形成。用细胞松弛素B处理后,pC5a或rhC5a可显著且同等程度地增强PMN-血小板悬液中TxB2的形成。在这些悬液中,A-23187诱导的血小板TxB2形成并未因PMN的存在而显著增加。A-23187诱导猪PMN产生显著的LXA4,添加血小板后并未增强。对PMN-血小板悬液的流式细胞术分析显示,pC5a刺激后活化的血小板黏附于PMN。血小板活化因子(PAF)受体拮抗剂CV-6209剂量依赖性地阻止pC5a诱导的血小板黏附于PMN及TxB2的形成。这些数据表明:1)猪PMN可单独生物合成LXA4,无需血小板协助,这与人类PMN-血小板相互作用形成鲜明对比;2)在无细胞松弛素B的情况下,pC5a刺激猪PMN合成PAF,导致血小板形成TxB2。