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Pharmacological and ischemic preconditioning of the human myocardium: mitoK(ATP) channels are upstream and p38MAPK is downstream of PKC.

Loubani M, Galiñanes M - BMC Physiol. (2002)

Bottom Line: Similar protection was obtained with alpha1 agonist phenylephrine, adenosine and IP and their combination did not afford additional cardioprotection.Furthermore, the protection induced by PMA was abolished by SB203580 but not by 5-hydroxydecanoate, whereas the protection induced by anisomycin was unaffected by either 5-hydroxydecanoate or chelerythrine.Opening of mitoK(ATP) channels and activation of PKC and p38MAPK are obligatory steps in the signal transduction cascade of IP and PP of the human myocardium with PKC activation being downstream of the opening of mitoK(ATP) channels and upstream of p38MAPK activation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Integrative Human Cardiovascular Physiology and Functional Genomics, Division of Cardiac Surgery, University of Leicester, Glenfield Hospital, Groby Road, Leicester. mahmoud.loubani@ntlworld.com

ABSTRACT

Background: These studies investigate the role of mitoK(ATP) channels, protein kinase C (PKC) and Mitogen activated protein kinase (p38MAPK) on the cardioprotection of ischemic (IP) and pharmacological preconditioning (PP) of the human myocardium and their sequence of activation.

Results: Right atrial appendages from patients undergoing elective cardiac surgery were equilibrated for 30 min and then subjected to 90 min of simulated ischemia followed by 120 min reoxygenation. At the end of each protocol creatinine kinase leakage (CK U/g wet wt) and the reduction of MTT to formazan dye (mM/g wet wt) were measured. Similar protection was obtained with alpha1 agonist phenylephrine, adenosine and IP and their combination did not afford additional cardioprotection. Blockade of mitoK(ATP) channels with 5-hydroxydecanoate, PKC with chelerythrine, or p38MAPK with SB203580 abolished the protection of IP and of PP. In additional studies, the stimulation of mitoK(ATP) channels with diazoxide or activation of PKC with PMA or p38MAPK with anisomycin induced identical protection to that of IP and PP. The protection induced by diazoxide was abolished by blockade of PKC and by blockade of p38MAPK. Furthermore, the protection induced by PMA was abolished by SB203580 but not by 5-hydroxydecanoate, whereas the protection induced by anisomycin was unaffected by either 5-hydroxydecanoate or chelerythrine.

Conclusions: Opening of mitoK(ATP) channels and activation of PKC and p38MAPK are obligatory steps in the signal transduction cascade of IP and PP of the human myocardium with PKC activation being downstream of the opening of mitoK(ATP) channels and upstream of p38MAPK activation.

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Protocol for Study 2 to investigate the role of PKC, p38MAPK and mitoKATP channels in the cardioprotective effect of IP and PP with phenylephrine (P) or adenosine (A) (n = 6 specimens/group): (i) time-matched aerobic control, (ii) SI/R alone, (iii) IP alone prior to SI, (iv) chelerythrine (CHE) for 10 min prior to IP, (v) SB203580 (SB) for 10 min prior to IP, (vi) 5-hydroxydecanoate (5-HD) for 10 min prior to IP, (vii) phenylephrine (P) for 5 min and 5 min washout (W) before SI, (viii) chelerythrine (CHE) for 10 min prior to phenylephrine (P) for 5 min and 5 min washout (W), (ix) SB203580 (SB) for 10 min prior to phenylephrine (P) for 5 min and 5 min washout (W), (x) 5-hydroxydecanoate (5-HD) for 10 min prior to phenylephrine (P) for 5 min and 5 min washout (W), (xi) adenosine (A) for 5 min and 5 min washout (W) before SI, (xii) chelerythrine (CHE) for 10 min prior to adenosine (A) for 5 min and 5 min washout (W), (xiii) SB203580 (SB) for 10 min prior to adenosine (A) for 5 min and 5 min washout (W) and (xiv) 5-hydroxydecanoate (5-HD) for 10 min prior to adenosine (A) for 5 min and 5 min washout (W).
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Figure 2: Protocol for Study 2 to investigate the role of PKC, p38MAPK and mitoKATP channels in the cardioprotective effect of IP and PP with phenylephrine (P) or adenosine (A) (n = 6 specimens/group): (i) time-matched aerobic control, (ii) SI/R alone, (iii) IP alone prior to SI, (iv) chelerythrine (CHE) for 10 min prior to IP, (v) SB203580 (SB) for 10 min prior to IP, (vi) 5-hydroxydecanoate (5-HD) for 10 min prior to IP, (vii) phenylephrine (P) for 5 min and 5 min washout (W) before SI, (viii) chelerythrine (CHE) for 10 min prior to phenylephrine (P) for 5 min and 5 min washout (W), (ix) SB203580 (SB) for 10 min prior to phenylephrine (P) for 5 min and 5 min washout (W), (x) 5-hydroxydecanoate (5-HD) for 10 min prior to phenylephrine (P) for 5 min and 5 min washout (W), (xi) adenosine (A) for 5 min and 5 min washout (W) before SI, (xii) chelerythrine (CHE) for 10 min prior to adenosine (A) for 5 min and 5 min washout (W), (xiii) SB203580 (SB) for 10 min prior to adenosine (A) for 5 min and 5 min washout (W) and (xiv) 5-hydroxydecanoate (5-HD) for 10 min prior to adenosine (A) for 5 min and 5 min washout (W).

Mentions: Creatine Kinase (CK) leakage into the media (A) during the 120 min reoxygenation period and MTT reduction by the slices (B) at the end of the reoxygenation period in human atrial myocardium subjected to various protocols (see Figure 2) to investigate the role of PKC, p38MAPK and mitoKATP channels in ischemic and pharmacological preconditioning with phenylephrine (P) or adenosine (Study 2). Data are expressed as mean ± SEM of six experiments. *p < 0.05 vs. SI/R alone group. (5-HD: 5-hydroxydecanoate, SB: SB203580).


Pharmacological and ischemic preconditioning of the human myocardium: mitoK(ATP) channels are upstream and p38MAPK is downstream of PKC.

Loubani M, Galiñanes M - BMC Physiol. (2002)

Protocol for Study 2 to investigate the role of PKC, p38MAPK and mitoKATP channels in the cardioprotective effect of IP and PP with phenylephrine (P) or adenosine (A) (n = 6 specimens/group): (i) time-matched aerobic control, (ii) SI/R alone, (iii) IP alone prior to SI, (iv) chelerythrine (CHE) for 10 min prior to IP, (v) SB203580 (SB) for 10 min prior to IP, (vi) 5-hydroxydecanoate (5-HD) for 10 min prior to IP, (vii) phenylephrine (P) for 5 min and 5 min washout (W) before SI, (viii) chelerythrine (CHE) for 10 min prior to phenylephrine (P) for 5 min and 5 min washout (W), (ix) SB203580 (SB) for 10 min prior to phenylephrine (P) for 5 min and 5 min washout (W), (x) 5-hydroxydecanoate (5-HD) for 10 min prior to phenylephrine (P) for 5 min and 5 min washout (W), (xi) adenosine (A) for 5 min and 5 min washout (W) before SI, (xii) chelerythrine (CHE) for 10 min prior to adenosine (A) for 5 min and 5 min washout (W), (xiii) SB203580 (SB) for 10 min prior to adenosine (A) for 5 min and 5 min washout (W) and (xiv) 5-hydroxydecanoate (5-HD) for 10 min prior to adenosine (A) for 5 min and 5 min washout (W).
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Figure 2: Protocol for Study 2 to investigate the role of PKC, p38MAPK and mitoKATP channels in the cardioprotective effect of IP and PP with phenylephrine (P) or adenosine (A) (n = 6 specimens/group): (i) time-matched aerobic control, (ii) SI/R alone, (iii) IP alone prior to SI, (iv) chelerythrine (CHE) for 10 min prior to IP, (v) SB203580 (SB) for 10 min prior to IP, (vi) 5-hydroxydecanoate (5-HD) for 10 min prior to IP, (vii) phenylephrine (P) for 5 min and 5 min washout (W) before SI, (viii) chelerythrine (CHE) for 10 min prior to phenylephrine (P) for 5 min and 5 min washout (W), (ix) SB203580 (SB) for 10 min prior to phenylephrine (P) for 5 min and 5 min washout (W), (x) 5-hydroxydecanoate (5-HD) for 10 min prior to phenylephrine (P) for 5 min and 5 min washout (W), (xi) adenosine (A) for 5 min and 5 min washout (W) before SI, (xii) chelerythrine (CHE) for 10 min prior to adenosine (A) for 5 min and 5 min washout (W), (xiii) SB203580 (SB) for 10 min prior to adenosine (A) for 5 min and 5 min washout (W) and (xiv) 5-hydroxydecanoate (5-HD) for 10 min prior to adenosine (A) for 5 min and 5 min washout (W).
Mentions: Creatine Kinase (CK) leakage into the media (A) during the 120 min reoxygenation period and MTT reduction by the slices (B) at the end of the reoxygenation period in human atrial myocardium subjected to various protocols (see Figure 2) to investigate the role of PKC, p38MAPK and mitoKATP channels in ischemic and pharmacological preconditioning with phenylephrine (P) or adenosine (Study 2). Data are expressed as mean ± SEM of six experiments. *p < 0.05 vs. SI/R alone group. (5-HD: 5-hydroxydecanoate, SB: SB203580).

Bottom Line: Similar protection was obtained with alpha1 agonist phenylephrine, adenosine and IP and their combination did not afford additional cardioprotection.Furthermore, the protection induced by PMA was abolished by SB203580 but not by 5-hydroxydecanoate, whereas the protection induced by anisomycin was unaffected by either 5-hydroxydecanoate or chelerythrine.Opening of mitoK(ATP) channels and activation of PKC and p38MAPK are obligatory steps in the signal transduction cascade of IP and PP of the human myocardium with PKC activation being downstream of the opening of mitoK(ATP) channels and upstream of p38MAPK activation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Integrative Human Cardiovascular Physiology and Functional Genomics, Division of Cardiac Surgery, University of Leicester, Glenfield Hospital, Groby Road, Leicester. mahmoud.loubani@ntlworld.com

ABSTRACT

Background: These studies investigate the role of mitoK(ATP) channels, protein kinase C (PKC) and Mitogen activated protein kinase (p38MAPK) on the cardioprotection of ischemic (IP) and pharmacological preconditioning (PP) of the human myocardium and their sequence of activation.

Results: Right atrial appendages from patients undergoing elective cardiac surgery were equilibrated for 30 min and then subjected to 90 min of simulated ischemia followed by 120 min reoxygenation. At the end of each protocol creatinine kinase leakage (CK U/g wet wt) and the reduction of MTT to formazan dye (mM/g wet wt) were measured. Similar protection was obtained with alpha1 agonist phenylephrine, adenosine and IP and their combination did not afford additional cardioprotection. Blockade of mitoK(ATP) channels with 5-hydroxydecanoate, PKC with chelerythrine, or p38MAPK with SB203580 abolished the protection of IP and of PP. In additional studies, the stimulation of mitoK(ATP) channels with diazoxide or activation of PKC with PMA or p38MAPK with anisomycin induced identical protection to that of IP and PP. The protection induced by diazoxide was abolished by blockade of PKC and by blockade of p38MAPK. Furthermore, the protection induced by PMA was abolished by SB203580 but not by 5-hydroxydecanoate, whereas the protection induced by anisomycin was unaffected by either 5-hydroxydecanoate or chelerythrine.

Conclusions: Opening of mitoK(ATP) channels and activation of PKC and p38MAPK are obligatory steps in the signal transduction cascade of IP and PP of the human myocardium with PKC activation being downstream of the opening of mitoK(ATP) channels and upstream of p38MAPK activation.

Show MeSH
Related in: MedlinePlus