Inhibition of superoxide anion generation by CHS-111 via blockade of the p21-activated kinase, protein kinase B/Akt and protein kinase C signaling pathways in rat neutrophils
Inhibition of superoxide anion generation by CHS-111 via blockade of the p21-activated kinase, protein kinase B/Akt and protein kinase C signaling pathways in rat neutrophils
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DOI：
10.1016/j.ejphar.2009.04.050
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发表时间：
2009-08-01
影响因子：
5
通讯作者：
Wang, Jih-Pyang
中科院分区：
文献类型：
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作者：
Chang, Ling-Chu;Lin, Ruey-Hseng;Wang, Jih-Pyang
在甲酰-Met-Leu-Phe（fMLP）刺激的大鼠中性粒细胞中，2-苄基-3-（4-羟甲基苯基）吲唑（CHS-111）可抑制超氧阴离子（O-2（中心点-））的产生，这不是通过清除产生的O-2（中心点-）或细胞毒性作用介导的，并减弱迁移。CHS-111对花生四烯酸诱导的NADPH氧化酶活化或无细胞系统中GTP γ S刺激的Rac 2膜转位没有影响，但它有效地减弱了p40（phox）、p47（phox）和p67（phox）的膜募集，p47 Ph（phox）中Set残基的磷酸化，p47（phox）和p22（phox）之间的结合，和fMLP刺激的嗜中性粒细胞中的Rac活化。CHS-111还能抑制p21激活激酶（PAK）的磷酸化和膜募集，抑制PAK激酶活性，抑制PAK与p47（phox）的相互作用。CHS-111有效降低Akt激酶活性和Akt与p47（phox）之间的关联，适度抑制Akt和磷酸化PDK 1的膜募集，并轻微减弱Akt（Thr 308）磷酸化，而对Akt（Ser 473）磷酸化或p110 γ膜转位无影响。蛋白激酶C（PKC）-α、-β I、β II、-δ和-zeta PKC磷酸化和PKC激酶活性的膜募集被CHS-111减弱，而CHS-111不影响p38丝裂原活化蛋白激酶（MAPK）或下游MAPK活化蛋白激酶-2的磷酸化。需要较高浓度的CHS-111来降低fMLP刺激的细胞内游离Ca 2+浓度（[Ca 2 +]（i））升高（在存在但不存在细胞外Ca 2+的情况下），并降低细胞环AMP但略微增加环GMP水平。综上所述，这些结果表明CHS-111通过阻断PAK、Akt和PKC信号通路抑制大鼠嗜中性粒细胞中fMLP刺激的O-2（中心点-）产生。(C)2009年爱思唯尔BIN。All rights reserved.
In formyl-Met-Leu-Phe (fMLP)-stimulated rat neutrophils, 2-benzyl-3-(4-hydroxymethylphenyl)indazole (CHS-111) inhibited superoxide anion (O-2(center dot-)) generation, which was not mediated by scavenging the generated O-2(center dot-) or by a cytotoxic effect, and attenuated migration. CHS-111 had no effect on the arachidonic acid-induced NADPH oxidase activation or the GTP gamma S-stimulated Rac2 membrane translocation in cell-free systems, whereas it effectively attenuated the membrane recruitment of p40(phox), p47(phox) and p67(phox), phosphorylation of Set residues in p47Ph(phox), association between p47(phox) and p22(phox), and Rac activation in fMLP-stimulated neutrophils. Moreover, the phosphorylation and membrane recruitment of p21-activated kinase (PAK), PAK kinase activity and the interaction of PAK with p47(phox) were inhibited by CHS-111. CHS-111 effectively reduced Akt kinase activity and the association between Akt and p47(phox), moderately inhibited the membrane recruitment of Akt and phospho-PDK1, and slightly attenuated Akt (Thr308) phosphorylation, whereas it had no effect on Akt (Ser473) phosphorylation or p110 gamma membrane translocation. The membrane recruitment of protein kinase C (PKC)-alpha, -beta I, beta II, -delta and -zeta PKC phosphorylation and PKC kinase activity was attenuated by CHS-111, whereas CHS-111 did not affect the phosphorylation of p38 mitogen-activated protein kinase (MAPK) or downstream MAPK-activated protein kinase-2. Higher concentrations of CHS-111 were required to decrease fMLP-stimulated intracellular free Ca2+ concentration ([Ca2+](i)) elevation in the presence but not in the absence of extracellular Ca2+, and to reduce cellular cyclic AMP but slightly increase cyclic GMP levels. Taken together, these results suggest that CHS-111 inhibits fMLP-stimulated O-2(center dot-) generation in rat neutrophils through the blockade of PAK, Akt and PKC signaling pathways. (C) 2009 Elsevier BIN. All rights reserved.