Highly effective non-viral gene transfer into vascular smooth muscle cells of cultured resistance arteries demonstrated by genetic inhibition of sphingosine-1-phosphate-induced vasoconstriction

The linkage of vascular genes to specific functions will lead to a
better understanding of cardiovascular pathophysiology. We
developed an experimental model that enables the introduction of
one or multiple gene(s) into vascular smooth muscle cells (VSMCs)
of isolated resistance arteries. Exposure of the arteries to a
green fluorescent protein (GFP)-encoding plasmid in combination
with the transfectant Effectene(R) for 20 h resulted in the
expression of GFP in virtually all VSMCs in the arterial wall at
fully preserved vascular function. For functional validation of the
model, plasmids encoding the specific RhoA inhibitors C3
transferase or N19RhoA were transfected. In subsequent functional
tests, inhibition of RhoA-dependent constriction induced by
sphingosine-1-phosphate was similar to that in arteries treated
with exogenous C3 transferase protein or the Rho kinase inhibitor
Y27632. Responses to norepinephrine remained unaffected. This novel
transfection technique enables gene function to be assessed in
direct conjunction with signalling pathways in vascular tissue and
provides, therefore, a new tool for microvascular proteomics.
Copyright (C) 2003 S. Karger AG, Basel.