Modification of the detrimental effect of TNF-α on human endothelial progenitor cells by fasudil and Y27632

Maria Luisa Balestrieri, Alfonso Giovane, Lara Milone, Francesca Felice, Carmela Fiorito, Valeria Crudele, Annaclaudia Esposito, Raffaele Rossiello, Pellegrino Biagio Minucci, Bartolomeo Farzati, Luigi Servillo, Claudio Napoli

Research output: Contribution to journalArticle

Abstract

Exposure of human endothelial progenitor cells (EPCs) to tumor necrosis factor-α (TNF-α) reduced their number and biological activity. Yet, signal transduction events linked to TNF-α action are still poorly understood. To address this issue, we examined the possible effect of fasudil and Y27632, two inhibitors of Rho kinase pathway, which is involved in endothelial dysfunction, atherosclerosis, and in- flammation. Results demonstrated that incubation with fasudil starting from 50 μM but not Y27632 determined a dose-dependent improvement of EPC number during exposure to TNF-α (P <0.05 vs. TNF-α alone). Analysis of the signal transduction pathway activated by TNF-α revealed that the increased expression of p-p38 was not significantly altered by fasudil. Instead, fasudil blocked the TNF-α induced phosphorylation of Erk1/2 (P <0.05 vs. TNF-α) as well as the inhibitor of Erk1/2-specific phosphorylated form, i.e., PD98059 (P <0.05 vs. TNF-α). These results were confirmed by analysis of these kinases by confocal microscopy. Finally, 2D-DIGE and MALDI-TOF/TOF analysis of EPCs treated with fasudil revealed increased expression levels of an actin-related protein and an adenylyl cyclase associated protein and decreased expression levels of proteins related to radical scavenger and nucleotide metabolism. These findings suggest that fasudil positively affects EPC number and that other major signals might take part to this complex pathway.

Original languageEnglish
Pages (from-to)351-360
Number of pages10
JournalJournal of Biochemical and Molecular Toxicology
Volume24
Issue number6
DOIs
Publication statusPublished - Nov 2010

Keywords

  • Endothelial progenitor cells
  • Fasudil
  • TNF-α

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Molecular Medicine
  • Toxicology
  • Health, Toxicology and Mutagenesis

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    Balestrieri, M. L., Giovane, A., Milone, L., Felice, F., Fiorito, C., Crudele, V., Esposito, A., Rossiello, R., Minucci, P. B., Farzati, B., Servillo, L., & Napoli, C. (2010). Modification of the detrimental effect of TNF-α on human endothelial progenitor cells by fasudil and Y27632. Journal of Biochemical and Molecular Toxicology, 24(6), 351-360. https://doi.org/10.1002/jbt.20345