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1 Department of Anatomy and Cell Biology and Cardiovascular Center, University of Iowa, Iowa City, Iowa 52242; and 2 Department of Medical Physiology, Texas A&M University, and System Health Science Center, College Station, Texas 77843
To test the hypotheses that cyclic stretch of
1) cardiac myocytes produces factors that trigger angiogenic
events in coronary microvascular endothelial cells (CMEC) and
2) CMEC enhances the expression of growth factors, cardiac
myocytes and CMEC were subjected to cyclic stretch in a
Flexercell Strain Unit. Vascular endothelial growth factor
(VEGF) but not basic fibroblast growth factor mRNA and protein levels
increased approximately twofold in myocytes after 1 h of stretch.
CMEC DNA synthesis increased approximately twofold when conditioned
medium from stretched myocytes or VEGF protein was added, and addition
of VEGF neutralizing antibody blocked the increase. CMEC migration and
tube formation increased with the addition of conditioned media but
were markedly attenuated by VEGF neutralizing antibody. Myocyte tumor
growth factor-
(TGF-) increased 2.5-fold after 1 h of
stretch, and the addition of TGF- neutralizing antibodies inhibited
the stretch-induced upregulation of VEGF. Stretch of CMEC increased
VEGF mRNA in these cells (determined by Northern blot and RT-PCR) and
increased the levels of VEGF protein (determined by ELISA analysis) in
the conditioned media. Therefore, cyclic stretch of cardiac myocytes
and CMEC appears to be an important primary stimulus for coronary
angiogenesis through both paracrine and autocrine VEGF pathways. These
data indicate that 1) CMEC DNA synthesis, migration, and
tube formation are increased in response to VEGF secreted from
stretched cardiac myocytes; 2) VEGF in CMEC subjected to
stretch is upregulated and secreted; and 3) TGF-
signaling may regulate VEGF expression in cardiac myocytes.
cardiac myocytes; endothelial cells; basic fibroblast growth
factor; cell migration; cell proliferation; vascular endothelial growth
factor; tumor growth factor-
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