Isoeugenodilol inhibits smooth muscle cell proliferation and neointimal thickening after balloon injury via inactivation of ERK1/2 pathway

Abstract

The purpose of this study was to determine the efficacy and the possible mechanism of action of the synthesized drug isoeugenodilol (a new third-generation beta-adrenoceptor blocker) on the growth factor-induced proliferation of cultured rat vascular smooth muscle cells (VSMCs) and neointimal formation in a rat carotid arterial balloon injury model. Isoeugenodilol significantly inhibited 10% FBS, 20 ng/ml PDGF-BB, and 20 ng/ml vascular endothelial growth factor (VEGF)-induced proliferation. In accordance with these findings, isoeugenodilol revealed blocking of the FBS-inducible progression through the G(0)/G(1) to the S phase of the cell cycle in synchronized cells. Neointimal formation, measured 14 days after injury, was reduced by the oral administration of isoeugenodilol (10 mg/kg/day). In an in vitro assay, isoeugenodilol inhibited the migration of VSMCs stimulated by PDGF-BB. These findings indicate that isoeugenodilol shows an inhibitory potency on neointimal formation due to inhibition of both migration and proliferation of VSMCs. In addition, isoeugenodilol in concentration-dependent manner decreased the levels of phosphorylated ERK1/2 in both VSMCs and balloon-injured carotid arteries. The levels of phosphorylated MEK1/2 and Pyk2 as well as intracellular Ca(2+) and reactive oxygen species (ROS) were in concentration-dependent manner reduced by isoeugenodilol. Taken together, these results indicate that isoeugenodilol may suppress mitogen-stimulated proliferation and migration partially through inhibiting cellular ROS and calcium, and hence, through activation of the Pyk2-ERK1/2 signal pathway. This suggests that isoeugenodilol has potential for the prevention of atherosclerosis and restenosis.

Induction of oxidative stress as a possible mechanism of the antifungal action of three phenylpropanoids

“…It is reasonable to envisage the involvement of oxidative stress in the antifungal activity of these phenylpropanoids, as the beneficial health effects of eugenol and related compounds are shown to be attributed mainly to free radical scavenging, metal chelating and modulating cell signalling pathways (Yeh et al , 2008; Kaur et al , 2010); however, the antimicrobial effects in this context are not fully characterized. Although all aerobically growing organisms are continuously exposed to reactive oxidants, oxidative stress occurs only when the concentration of the oxidants increases beyond the antioxidant capacity of the cells (Fridovich, 1998).…”

Gamma-secretase Inhibitor Prevents Proliferation and Migration of Ductus Arteriosus Smooth Muscle Cells through the Notch3-HES1/2/5 Pathway

“…In order to measure ROS production, cells were stained with 10 μM of DCFH-DA for 30 min at 37℃, detached with trypsin/EDTA, washed, re-suspended in PBS, and then immediately analyzed by flow cytometry. A FACScan flow cytometer was used to measure ROS generation by the fluorescence intensity (FL-1, 525 nm) of 10,000 cells as we previously described 23.…”

“…The protein samples were extracted from the whole cells with the procedures as we previously described 22, 23. All of the extracted protein solutions were stored at -80℃ until analysis.…”

Hydrogen-rich saline attenuates vascular smooth muscle cell proliferation and neointimal hyperplasia by inhibiting reactive oxygen species production and inactivating the Ras-ERK1/2-MEK1/2 and Akt pathways

“…Histologically stained sections were used to assess the extent of neointimal formation using computed planimetry. The intima-to-media (I/M) area ratio was calculated as the mean of these determinations (23).…”

“…VSMCs (1x10 5 cells) were incubated with 10% FBS in the presence or absence of HRM for 12 h. Cells were stained with 10 µM of DCFH-DA for 30 min at 37˚C, detached with trypsin/EDTA, washed, re-suspended in PBS, and immediately analyzed by flow cytometry using a FACScan by fluorescence intensity (FL-1,525 nm) of 10,000 cells (23).…”