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Ethanol inhibits mitogen activated protein kinase activity and growth of vascular smooth muscle cells in vitro
R J Hendrickson1, P A Cahill, I H McKillop
1Georgetown University Medical Center, Department of Surgery, Pasquerilla Healthcare Center, Washington, DC 20007, USA.
Abstract:
The aim of this study was to determine the effect of ethanol on vascular smooth muscle cell proliferation and mitogen activated protein kinase (MAPK) signaling. Rat aortic smooth muscle cell growth in vitro was determined by measuring cell counts and [3H]thymidine incorporation. MAPK signaling was determined by assessing MEK (also referred to as MAPK kinase) activity by measuring phosphorylated extracellular signal-regulated kinase (pp44ERK - 1 and pp42ERK - 2) expression, and ERK activity by measuring ERK-2-dependent phosphorylation of myelin basic protein (MBP). In quiesced smooth muscle cells, ethanol treatment (24 h) inhibited serum-stimulated mitogenesis in a dose-dependent manner, (IC50 = 60 mM), in the absence of any effect on smooth muscle cell viability. In addition, ethanol treatment caused a significant shift to the right in the smooth muscle cell growth curve, extending the population doubling time from approximately 48 h (control) to approximately 70 h (ethanol). Acute (15 min) ethanol treatment reduced serum-stimulated pp44ERK - 1 and pp42ERK - 2 expression in a dose dependent fashion; 24.5+/-1.5% and 77.6+/-3.2% inhibition for 20 mM and 160 mM ethanol, respectively. Furthermore, there was a significant dose-dependent decrease in ERK2 activity in ethanol treated smooth muscle cells as compared to control smooth muscle cells. These data demonstrate an inhibitory effect of ethanol on smooth muscle cell proliferation and MAPK signalling in vitro. It is tempting to speculate that these actions of ethanol may contribute to its cardiovascular effects in vivo.
Insights
Ethanol inhibits vascular smooth muscle cell proliferation and mitogen-activated protein kinase (MAPK) signaling in vitro. This study investigated ethanol
Area of Science:
- Vascular Biology
- Cell Signaling
- Pharmacology
Background:
- Vascular smooth muscle cells (VSMCs) are crucial for regulating blood pressure and vessel tone.
- Mitogen-activated protein kinase (MAPK) pathways are key regulators of cell proliferation and survival.
- Ethanol's cardiovascular effects are complex and not fully understood at the cellular level.
Purpose of the Study:
- To investigate the impact of ethanol on VSMC proliferation.
- To elucidate the effect of ethanol on MAPK signaling pathways in VSMCs.
- To determine if ethanol affects VSMC viability.
Main Methods:
- Rat aortic smooth muscle cells were cultured in vitro.
- Cell proliferation was assessed by cell counts and [3H]thymidine incorporation.
- MAPK signaling was analyzed by measuring MEK activity, ERK phosphorylation (pp44ERK and pp42ERK), and ERK2 activity.
Main Results:
- Ethanol inhibited serum-stimulated VSMC mitogenesis in a dose-dependent manner (IC50 = 60 mM) without affecting cell viability.
- Ethanol treatment extended the population doubling time of VSMCs from 48 to 70 hours.
- Acute ethanol exposure dose-dependently reduced ERK phosphorylation and ERK2 activity.
Conclusions:
- Ethanol exhibits an inhibitory effect on vascular smooth muscle cell proliferation in vitro.
- Ethanol interferes with MAPK signaling pathways in VSMCs.
- These cellular effects of ethanol may contribute to its in vivo cardiovascular actions.