TY - JOUR
T1 - Synergistic modulation of ATP-sensitive K+ currents by protein kinase C and adenosine
T2 - Implications for ischemic preconditioning
AU - Liu, Yongge
AU - Gao, Wei Dong
AU - O'Rourke, Brian
AU - Marban, Eduardo
PY - 1996/3
Y1 - 1996/3
N2 - Ischemic preconditioning has been shown to involve the activation of adenosine receptors, protein kinase C (PKC), and ATP sensitive K+ (K(ATP)) channels. We investigated the effects of PKC activation and adenosine on K(ATP) current (I(KATP))) and action potentials in isolated rabbit ventricular myocytes. Responses to pinacidil (100 to 400 μmol/L), an opener of K(ATP) channels, were markedly increased by preexposure to the PKC activator phorbol 12-myristate 13-acetate (PMA, 100 nmol/L). I(KATP) measured at 0 mV was increased by PMA pretreatment from 0.55±0.32 to 3.25±0.47 nA (n=6, P<.01). We next determined whether PKC activation abbreviates the time required to turn on I(KATP) during metabolic inhibition (MI). In control cells in which MI was induced by 2 mmol/L cyanide and 0 glucose, I(KATP) developed after an average of 15.1±2.4 minutes (n=8). Ten-minute pretreatment with PMA alone (PMA+MI) did not significantly alter this latency (11.9±2.0 minutes, n=8). Since adenosine receptor activation has been shown to play an important role in the preconditioning response, two groups of myocytes were studied with adenosine (10 μmol/L) included during MI. Without PMA, adenosine alone (MI+Ado) did not affect the latency to develop I(KATP) (12.3±1.5 minutes, n=8). However, if cells were pretreated with PMA and then subjected to MI in the presence of adenosine (PMA+MI+Ado), the latency was greatly shortened to 5.5±1.6 minutes (n=8; P<.02 versus MI, PMA+MI, and MI+Ado groups). This effect could not bc reproduced by an inactive phorbol but was completely abolished by the adenosine receptor antagonist 8-(p- sulfophenyl)-theophylline. The opening of K(ATP) channels may be cardioprotective because of the abbreviation of action potential duration (APD) during ischemia. Therefore, we tested whether PKC activation could modify the time course of APD shortening during MI. Consistent with the ionic current measurements, PMA pretreatment significantly accelerated APD shortening, but only when adenosine (10 μmol/L) was included during MI. The effects were not attributable to accelerated ATP consumption: PMA pretreatment did not alter the time required to induce rigor during MI, whether or not adenosine was included. Our results indicate that PKC activation increases the I(KATP) induced by pinacidil or by MI. The latter effect requires concomitant adenosine receptor activation. The synergistic modulation of I(KATP) by PKC and adenosine provides an explicit basis for current paradigms of ischemic preconditioning.
AB - Ischemic preconditioning has been shown to involve the activation of adenosine receptors, protein kinase C (PKC), and ATP sensitive K+ (K(ATP)) channels. We investigated the effects of PKC activation and adenosine on K(ATP) current (I(KATP))) and action potentials in isolated rabbit ventricular myocytes. Responses to pinacidil (100 to 400 μmol/L), an opener of K(ATP) channels, were markedly increased by preexposure to the PKC activator phorbol 12-myristate 13-acetate (PMA, 100 nmol/L). I(KATP) measured at 0 mV was increased by PMA pretreatment from 0.55±0.32 to 3.25±0.47 nA (n=6, P<.01). We next determined whether PKC activation abbreviates the time required to turn on I(KATP) during metabolic inhibition (MI). In control cells in which MI was induced by 2 mmol/L cyanide and 0 glucose, I(KATP) developed after an average of 15.1±2.4 minutes (n=8). Ten-minute pretreatment with PMA alone (PMA+MI) did not significantly alter this latency (11.9±2.0 minutes, n=8). Since adenosine receptor activation has been shown to play an important role in the preconditioning response, two groups of myocytes were studied with adenosine (10 μmol/L) included during MI. Without PMA, adenosine alone (MI+Ado) did not affect the latency to develop I(KATP) (12.3±1.5 minutes, n=8). However, if cells were pretreated with PMA and then subjected to MI in the presence of adenosine (PMA+MI+Ado), the latency was greatly shortened to 5.5±1.6 minutes (n=8; P<.02 versus MI, PMA+MI, and MI+Ado groups). This effect could not bc reproduced by an inactive phorbol but was completely abolished by the adenosine receptor antagonist 8-(p- sulfophenyl)-theophylline. The opening of K(ATP) channels may be cardioprotective because of the abbreviation of action potential duration (APD) during ischemia. Therefore, we tested whether PKC activation could modify the time course of APD shortening during MI. Consistent with the ionic current measurements, PMA pretreatment significantly accelerated APD shortening, but only when adenosine (10 μmol/L) was included during MI. The effects were not attributable to accelerated ATP consumption: PMA pretreatment did not alter the time required to induce rigor during MI, whether or not adenosine was included. Our results indicate that PKC activation increases the I(KATP) induced by pinacidil or by MI. The latter effect requires concomitant adenosine receptor activation. The synergistic modulation of I(KATP) by PKC and adenosine provides an explicit basis for current paradigms of ischemic preconditioning.
KW - ATP-sensitive
KW - K current
KW - adenosine
KW - ischemic preconditioning
KW - pinacidil
KW - protein kinase C
UR - http://www.scopus.com/inward/record.url?scp=9044246680&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=9044246680&partnerID=8YFLogxK
U2 - 10.1161/01.RES.78.3.443
DO - 10.1161/01.RES.78.3.443
M3 - Article
C2 - 8593703
AN - SCOPUS:9044246680
SN - 0009-7330
VL - 78
SP - 443
EP - 454
JO - Circulation research
JF - Circulation research
IS - 3
ER -