Actin polymerization, calcium-transients, and phospholipid metabolism in human neutrophils after stimulation with interleukin-8 and N-formyl peptide.
Actin polymerization, calcium-transients, and phospholipid metabolism in human neutrophils after stimulation with interleukin-8 and N-formyl peptide.
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用 IL-8 和 N-甲酰肽刺激后人中性粒细胞中的肌动蛋白聚合、钙瞬变和磷脂代谢。
DOI：
10.1111/1523-1747.ep12371788
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发表时间：
1994
期刊：
影响因子：
--
通讯作者：
Schraufstätter,IU
中科院分区：
文献类型：
--
作者：
Norgauer,J;Krutmann,J;Dobos,GJ;Traynor-Kaplan,AE;Oades,ZG;Schraufstätter,IU
对中性粒细胞中白细胞介素 8 (IL-8) 的信号转导进行了分析，并与众所周知的中性粒细胞激活剂 N-甲酰肽进行比较。用 IL-8 刺激人中性粒细胞诱导肌动蛋白快速聚合，通过 f-肌动蛋白的 7-硝基苯甲-2-氧杂-1,3-二唑-(NBD)-鬼笔酸丁染色检测到，并减少监测到的直角光散射。肌动蛋白聚合在添加 IL-8 后 10 秒内达到峰值，与 N-甲酰肽诱导的刺激相比，持续时间较短。通过薄层色谱分析磷脂并通过高压液相色谱 (HPLC) 分析脂质提取物的脱酰化产物表明，IL-8 引发 [32P] 磷脂酰肌醇 (3,4,5) 三磷酸 (PtdInsP3) 快速升高，随后缓慢升高[32P]磷脂酰肌醇(3,4)二磷酸(Ptdlns-3,4-P2)随着[32P]磷脂酰肌醇(4,5)二磷酸(Ptdlns-4,5-P2)的快速减少。多磷酸肌醇代谢的变化比 N-甲酰肽获得的变化更为温和和短暂。此外，与 N-甲酰肽激活的反应相比，IL-8 刺激的 [32P]磷脂酸 (PA) 产生是最小且短暂的。IL-8 和 N-甲酰肽均诱导细胞内储存的 Ca++ 动员，但与 N-甲酰肽相比，IL-8 未能触发 Ca++ 从细胞外介质的二次流入。总之，IL-8 和 N-甲酰肽刺激相似且不同的细胞内激活步骤模式。这项研究表明，IL-8 是趋化性所需的细胞内事件的有效激活剂，但对于与超氧阴离子生成和促炎活性相关的事件的激活剂相对较弱。
Signal transduction of interleukin-8 (IL-8) was analyzed in neutrophils, and compared with the well known neutrophil activator N-formyl peptide. Stimulation of human neutrophils with IL-8 induced a rapid polymerization of actin as detected by 7-nitrobenz-2-oxa-1,3-diazol-(NBD)-phallacidin staining of f-actin and reduction of monitored right-angle light scatter. Actin polymerization peaked within 10 seconds after the addition of IL-8 and was short-lived as compared to N-formyl peptide-induced stimulation.Analysis of phospholipids by thin-layer chromatography and analysis of deacylation products of lipid extracts by high-pressure liquid chromatography (HPLC) showed that IL-8 triggered a rapid rise of [32P]phosphatidyl-inositol(3,4,5)trisphosphate (PtdInsP3) followed by a slower increase of [32P]phosphatidyl-inositol(3,4)bisphosphate (Ptdlns-3,4-P2) along with a rapid decrease of [32P]phosphatidylinositol(4,5)bisphosphate (Ptdlns-4,5-P2). Changes in polyphos-phoinositide metabolism were more moderate and transient than those obtained by N-formyl peptide. Moreover, [32P]phosphatidic acid (PA) production stimulated by IL-8 was minimal and transient as compared to the response activated by N-formyl peptide.Both IL-8 and N-formyl peptide induced Ca++mobilization from intracellular stores, but IL-8 in contrast to N- formyl peptide failed to trigger the secondary influx of Ca++from the extracellular medium. In summary, IL-8 and N-formyl peptide stimulated similar and distinct patterns of intracellular activation steps. This study indicates that IL-8 is a potent activator of intracellular events presumably required for chemotaxis, but a relatively weak activator for events associated with superoxide anion generation and proinflammatory activity.