对大鼠中性粒细胞中由甲酰甲硫氨酰亮氨酰苯丙氨酸导致细胞外信号调节激酶激活的信号转导途径的研究。

Examination of the signal transduction pathways leading to activation of extracellular signal-regulated kinase by formyl-methionyl-leucyl-phenylalanine in rat neutrophils.

作者信息

Chang L C, Wang J P

机构信息

Department of Education and Research, Taichung Veterans General Hospital, Taiwan, ROC.

出版信息

FEBS Lett. 1999 Jul 2;454(1-2):165-8. doi: 10.1016/s0014-5793(99)00717-6.

DOI:10.1016/s0014-5793(99)00717-6

PMID:10413116

Abstract

The signaling pathways leading to extracellular signal-regulated kinase (ERK) activation in formyl-methionyl-leucyl-phenylalanine (fMLP)-stimulated rat neutrophils were examined. fMLP-stimulated ERK activation based on immunoblot analysis with antibodies against the phosphorylation form of ERK was attenuated by the pretreatment of cells with pertussis toxin but not with a dual cyclo-oxygenase/lipoxygenase inhibitor BW755C. Exposure of cells to the tyrosine kinase inhibitor genistein, phosphatidylinositol 3-kinase (PI3K) inhibitors wortmannin and LY294002, or protein kinase C (PKC) inhibitors Gö6976, Gö6983, and GF109203X inhibited fMLP-stimulated ERK phosphorylation in a concentration-dependent manner. In addition, both the phospholipase C (PLC) inhibitor U73122 and the Ca2+ chelator BAPTA attenuated ERK activation. These results indicate that G(i/o) protein, tyrosine kinase, P13K, PKC, and PLC/Ca2+, but not arachidonate metabolites, act upstream of fMLP-stimulated ERK activation.

摘要

研究了在甲酰甲硫氨酰亮氨酰苯丙氨酸（fMLP）刺激的大鼠中性粒细胞中导致细胞外信号调节激酶（ERK）激活的信号通路。基于用抗ERK磷酸化形式抗体进行免疫印迹分析，fMLP刺激的ERK激活被百日咳毒素预处理细胞所减弱，但双环氧化酶/脂氧合酶抑制剂BW755C预处理则无此作用。细胞暴露于酪氨酸激酶抑制剂染料木黄酮、磷脂酰肌醇3激酶（PI3K）抑制剂渥曼青霉素和LY294002，或蛋白激酶C（PKC）抑制剂Gö6976、Gö6983和GF109203X，均以浓度依赖性方式抑制fMLP刺激的ERK磷酸化。此外，磷脂酶C（PLC）抑制剂U73122和Ca2+螯合剂BAPTA均减弱了ERK激活。这些结果表明，G(i/o)蛋白、酪氨酸激酶、PI3K、PKC和PLC/Ca2+，而非花生四烯酸代谢产物，在fMLP刺激的ERK激活上游起作用。