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

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.

The Rho kinase inhibitor fasudil augments the number of functional endothelial progenitor cells in ex vivo cultures

“…However, rocK inhibitors may be beneficial in EPC culture, since only the more selective ROCK inhibitors effectively enhanced the number of ePcs. another group has reported that fasudil modifies the detrimental effect of tnf-α on human peripheral blood ePcs by recovering ePc number (24), supporting our notion that fasudil positively affects ePc number. However, in a previous study, treatment of fasudil for 24 h did not noticeably augment ePc number when mncs from the peripheral blood of healthy human volunteers were treated (16).…”

Involvement of endothelial progenitor cells in the formation of plexiform lesions in broiler chickens: possible role of local immune/inflammatory response

“…We therefore used a cell co-culture system consisting of EPCs and lymphocytes at a ratio of 1:1 to test our hypothesis. We believed that our co-culture system would be more realistic to mimic the in vivo events than stimulating the EPC with a single type of proinflammatory cytokine (Balestrieri et al, 2010;Zhao et al, 2012). The EPC population used in this work was composed of so-called "late" EPCs, which have proliferative capacity and are able to differentiate into mature endothelial cells (Hur et al, 2004).…”

Identification of the Transmembrane Glucose Regulated Protein 78 as a Biomarker for the Brain Cancer Glioblastoma Multiforme by Gene Expression and Proteomic Studies.

“…In particular, 2D-DIGE utilizes mass and charge-matched spectrally resolvable fluorescent dyes (Cy3 and Cy5) to label two different protein samples in vitro prior to two-dimensional electrophoresis. To date, 2D/DIGE is still one of the central technologies in proteomics for the separation and differential comparison of thousands of proteins in a complex mixture [ 21 - 23 ].…”