(Circulation. 1995;91:2226-2234.)
© 1995 American Heart Association, Inc.
Articles |
1-Adrenoceptor Activation Increases Ecto-5'-Nucleotidase Activity and Adenosine Release in Rat Cardiomyocytes by Activating Protein Kinase C
From the First Department of Medicine, Osaka University School of Medicine, Suita; Cardiology, Department of Internal Medicine, Sakurabashi Watanabe Hospital, Osaka (K.I.); and the Department of Medical Information Science, Osaka University Hospital, Suita (M.I.), Japan.
Background Adenosine is an important regulator of many
cardiac functions and is synthesized primarily by ecto- and cytosolic
5'-nucleotidase. We have previously reported that
1-adrenoceptor blockade attenuates adenosine release
from ischemic myocardium, raising the possibility that
1-adrenoceptor activation activates 5'-nucleotidase.
This study tested whether activation of protein kinase C by
1-adrenoceptor activation increases 5'-nucleotidase
activity and augments adenosine release.
Methods and Results Cardiomyocytes were isolated from adult male
Wistar rats and suspended in modified HEPES-Tyrode's buffer solution.
After stabilization, the cardiomyocytes were incubated with and without
an exposure to norepinephrine (10-9 to 10-5
mol/L) while being treated with propranolol and yohimbine or with and
without an exposure to methoxamine (10-9 to
10-5 mol/L). Ecto-5'-nucleotidase activity was
increased
by norepinephrine and methoxamine during 30 minutes in a dose-dependent
manner, whereas cytosolic 5'-nucleotidase was not activated. These
increases in ecto-5'-nucleotidase activity were inhibited by
GF109203X, an inhibitor of protein kinase C, and mimicked by phorbol
12-myristate 13-acetate (PMA), an activator of protein kinase C. The
increase in ecto-5'-nucleotidase was not prevented by cycloheximide.
When ecto-5'-nucleotidase activity increased, adenosine release was
augmented in methoxamine- and PMA-treated cardiomyocytes (1299±252%
and 1372±149%, respectively) compared with the untreated group
(578±26%). The increase in adenosine release was blunted by GF109203X
and ,ß-methyleneadenosine 5'-diphosphate, an inhibitor of
ecto-5'-nucleotidase.
Conclusions Thus, we conclude that
1-adrenoceptor–mediated increases in
ecto-5'-nucleotidase activity are attributed to activation of protein
kinase C in rat cardiomyocytes.
Key Words: nucleotides • norepinephrine • ischemia • hypoxia
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