Dissociation of the effects of forskolin and dibutyryl cAMP on force of contraction and phospholamban phosphorylation in human heart failure.
Forskolin and dibutyryl cyclic adenosine monophosphate (cAMP) stimulate force of contraction independent of beta-adrenoceptor stimulation. We studied their effects on force of contraction and phosphorylation of regulatory proteins in isolated electrically driven trabeculae carneae from failing human ventricles. The phosphorylation state of the regulatory protein phospholamban was studied because its phosphorylation usually faithfully follows contractility. For comparison, the phosphorylation state of the inhibitory subunit of troponin was studied. The phosphorylation state was inferred from in vitro phosphorylation of homogenates with cAMP-dependent protein kinase in the presence of radioactive gamma[32P]ATP Proteins were separated by electrophoresis, and radioactivity in the proteins of interest was quantified. The maximal positive inotropic effects occurred at 30 microM forskolin and were attenuated in comparison with the maximal effects to dibutyryl cAMP (1 mM). Both forskolin and dibutyryl cAMP enhanced phospholamban phosphorylation. However, phospholamban phosphorylation in intact trabeculae treated with 30 microM forskolin and 1 mM dibutyryl cAMP was comparable. It is suggested that phospholamban phosphorylation can be dissociated from inotropy at least in isolated trabeculae from failing human hearts.
Neumann
J
J
Institut für Pharmakologie und Toxikologie, Universität Münster, FRG.
Bartel
S
S
Eschenhagen
T
T
Haverich
A
A
Hirt
S
S
Karczewski
P
P
Krause
E G
EG
Schmitz
W
W
Scholz
H
H
Stein
B
B
Thoenes
M
M
D000293:Adolescent; D000328:Adult; D000368:Aged; D003994:Bucladesine / Q000494:pharmacology*; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D002316:Cardiotonic Agents; D005576:Colforsin / Q000494:pharmacology*; D005260:Female; D006333:Heart Failure / Q000503:physiopathology*; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D008875:Middle Aged; D009200:Myocardial Contraction / Q000187:drug effects*; D010766:Phosphorylation; D000099211:Phospholamban
D003994:Bucladesine; D002135:Calcium-Binding Proteins; D002316:Cardiotonic Agents; D005576:Colforsin; D000099211:Phospholamban