Actin reorganization and morphological changes in human neutrophils stimulated by TNF, GM-CSF, and G-CSF: the role of MAP kinases
Actin reorganization and morphological changes in human neutrophils stimulated by TNF, GM-CSF, and G-CSF: the role of MAP kinases
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DOI：
10.1152/ajpcell.00131.2003
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发表时间：
2004-01-01
影响因子：
5.5
通讯作者：
Kitagawa, S
中科院分区：
文献类型：
--
作者：
Kutsuna, H;Suzuki, K;Kitagawa, S
肿瘤坏死因子- α (TNF)、粒细胞-巨噬细胞集落刺激因子（GM-CSF）或粒细胞CSF （G-CSF）刺激人中性粒细胞可导致FITC-phalloidin（肌动蛋白解聚）荧光强度降低和形态改变。细胞因子诱导的肌动蛋白解聚取决于用作刺激的细胞因子的浓度。在TNF或GM-CSF刺激后10分钟和G-CSF刺激后20分钟检测到最大变化。刺激后细胞因子诱导的肌动蛋白解聚持续至少30分钟。相反，n -甲酰基-蛋氨酸-亮基-苯丙氨酸（FMLP）快速（45 s内）诱导FITC-phalloidin（肌动蛋白聚合）荧光强度增加和形态改变。PD-98059(丝裂原活化蛋白激酶（MAPK)/细胞外信号调节激酶（ERK）激酶抑制剂）或SB-203580 （p38 MAPK抑制剂）可部分抑制TNF-和gm - csf诱导的肌动蛋白解聚和形态变化，但fmlp诱导的反应未被抑制，并且这些抑制剂联合使用几乎完全消除。PD-98059几乎完全消除了g - csf诱导的反应，SB-203580则不受影响。这些发现与这些细胞因子激活人类中性粒细胞中独特的MAPK亚型级联的能力是一致的。在细胞因子或FMLP刺激的中性粒细胞中，磷酸化的ERK和p38 MAPK未与f -肌动蛋白共定位。此外，细胞因子预处理可防止fmlp诱导的极化和肌动蛋白聚合。这些发现表明，TNF、GM-CSF和G-CSF通过激活ERK和/或p38 MAPK诱导肌动蛋白解聚和形态学改变，细胞因子诱导的肌动蛋白重组可能是这些细胞因子抑制中性粒细胞趋化作用的部分原因。
Stimulation of human neutrophils with tumor necrosis factor-alpha (TNF), granulocyte-macrophage colony-stimulating factor (GM-CSF), or granulocyte CSF (G-CSF) resulted in decreased fluorescence intensity of FITC-phalloidin ( actin depolymerization) and morphological changes. Cytokine-induced actin depolymerization was dependent on the concentration of cytokines used as stimuli. The maximal changes were detected at 10 min after stimulation with TNF or GM-CSF and at 20 min after stimulation with G-CSF. Cytokine-induced actin depolymerization was sustained for at least 30 min after stimulation. In contrast, N-formyl-methionyl-leucyl-phenylalanine ( FMLP) rapidly ( within 45 s) induced an increase in the fluorescence intensity of FITC-phalloidin ( actin polymerization) and morphological changes. TNF- and GM-CSF-induced actin depolymerization and morphological changes, but not FMLP-induced responses, were partially inhibited by either PD-98059, an inhibitor of mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) kinase, or SB-203580, an inhibitor of p38 MAPK, and were almost completely abolished by these inhibitors in combination. G-CSF-induced responses were almost completely abolished by PD-98059 and were unaffected by SB-203580. These findings are consistent with the ability of these cytokines to activate the distinct MAPK subtype cascade in human neutrophils. Phosphorylated ERK and p38 MAPK were not colocalized with F-actin in neutrophils stimulated by cytokines or FMLP. Furthermore, FMLP-induced polarization and actin polymerization were prevented by cytokine pretreatment. These findings suggest that TNF, GM-CSF, and G-CSF induce actin depolymerization and morphological changes through activation of ERK and/or p38 MAPK and that cytokine-induced actin reorganization may be partly responsible for the inhibitory effect of these cytokines on neutrophil chemotaxis.