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
| Autor: | Li-Jiau Huang, Sheng-Chu Kuo, Ruey-Hseng Lin, Ling-Chu Chang, Chi-Sen Chang, Jih-Pyang Wang |
|---|---|
| Rok vydání: | 2009 |
| Předmět: |
Indazoles
Protein Kinase C-alpha Neutrophils Biology p38 Mitogen-Activated Protein Kinases Protein kinase C signaling Rats Sprague-Dawley Superoxides Cyclic AMP Animals Phosphorylation Kinase activity Protein kinase A Cyclic GMP Protein kinase B Protein Kinase C Protein kinase C Pharmacology Cell-Free System Kinase NADPH Oxidases Molecular biology Rats Cell biology p21-Activated Kinases Signal transduction Proto-Oncogene Proteins c-akt Signal Transduction |
| Zdroj: | European Journal of Pharmacology. 615:207-217 |
| ISSN: | 0014-2999 |
| DOI: | 10.1016/j.ejphar.2009.04.050 |
| Popis: | In formyl-Met-Leu-Phe (fMLP)-stimulated rat neutrophils, 2-benzyl-3-(4-hydroxymethylphenyl)indazole (CHS-111) inhibited superoxide anion (O(2)(-)) generation, which was not mediated by scavenging the generated O(2)(-) or by a cytotoxic effect, and attenuated migration. CHS-111 had no effect on the arachidonic acid-induced NADPH oxidase activation or the GTPgammaS-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 Ser residues in p47(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 p110gamma membrane translocation. The membrane recruitment of protein kinase C (PKC)-alpha, -betaI, -betaII, -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 Ca(2+) concentration ([Ca(2+)](i)) elevation in the presence but not in the absence of extracellular Ca(2+), 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)(-) generation in rat neutrophils through the blockade of PAK, Akt and PKC signaling pathways. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect