(Circulation. 1999;99:2523-2529.)
© 1999 American Heart Association, Inc.
Clinical Investigation and Reports |
Calcium Antagonists Ameliorate Ischemia-Induced Endothelial Cell Permeability by Inhibiting Protein Kinase C
From the Franz Volhard Clinic and Max Delbrück Center for Molecular Medicine (A.H., C.L., C.M., M.M., R.B., F.C.L., H.H.), Medizinische Fakultät der Charité, Humboldt University of Berlin, and Physiologisches Institut (T.N.), Justus Liebig University, Giessen, Germany.
Correspondence to Hermann Haller, MD, Franz Volhard Clinic, Wiltberg Strasse 50, 13122 Berlin, Germany. E-mail haller{at}fvk-berlin.de
Background—Dihydropyridines block calcium channels; however, they also influence endothelial cells, which do not express calcium channels. We tested the hypothesis that nifedipine can prevent ischemia-induced endothelial permeability increases by inhibiting protein kinase C (PKC) in cultured porcine endothelial cells.
Methods and Results—Ischemia was induced by potassium
cyanide/deoxyglucose, and permeability was measured by albumin
flux. Ion channels were characterized by patch clamp.
[Ca2+]i was measured by fura 2. PKC activity
was measured by substrate phosphorylation after cell
fractionation. PKC isoforms were assessed by Western blot and confocal
microscopy. Nifedipine prevented the
ischemia-induced increase in permeability in a dose-dependent
manner. Ischemia increased [Ca2+]i,
which was not affected by nifedipine. Instead,
ischemia-induced PKC translocation was prevented by
nifedipine. Phorbol ester also increased
endothelial cell permeability, which was dose
dependently inhibited by nifedipine. The effects of
non–calcium-channel–binding dihydropyridine
derivatives were similar. Analysis of the PKC isoforms showed
that nifedipine prevented ischemia-induced
translocation of PKC-
and PKC-
. Specific inhibition of PKC
isoforms with antisense oligodeoxynucleotides demonstrated
a major role for PKC-.
Conclusions—Nifedipine exerts a direct effect on
endothelial cell permeability that is independent of
calcium channels. The inhibition of ischemia-induced
permeability by nifedipine seems to be mediated primarily
by PKC- inhibition. Anti-ischemic effects of
dihydropyridine calcium antagonists
could be due in part to their effects on endothelial
cell permeability.
Key Words: cells • ischemia • proteins • calcium
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