The activation of p38alpha, and not p38beta, mitogen-activated protein kinase is required for ischemic preconditioning

Abstract 

Numerous studies show that pharmacological inhibition of p38 mitogen-activated protein kinases (p38s) before lethal ischemia prevents conditioning. However, these inhibitors have off-target effects and do not discriminate between the alpha and beta isoforms; the activation of which is thought to have diverse and perhaps opposing actions with p38α aggravating, and p38β reducing, myocardial injury. We adopted a chemical genetic approach using mice in which either the p38α (DRα) or p38β (DRβ) alleles were targeted to substitute the “gatekeeper” threonine residue for methionine, thereby preventing the binding of a pharmacological inhibitor, SB203580. Isolated, perfused wild-type (WT), DRα and DRβ mouse hearts underwent ischemic preconditioning with 4cycles of 4min ischemia/6min reperfusion, with or without SB203580 (10µM), followed by 30min of global ischemia and 120min of reperfusion. In WT and DRβ hearts, SB203580 completely abolished the reduction in myocardial infarction seen with preconditioning and also the phosphorylation of downstream substrates of p38. These effects of SB203580 were not seen in DRα hearts. Furthermore ischemic preconditioning occurred unaltered in p38β null hearts. Contrary to expectation the activation of p38α, and not p38β, is necessary for ischemic preconditioning. Since p38α is also the isoform that leads to lethal myocardial injury, it is unlikely that targeted therapeutic strategies to achieve isoform-selective inhibition will only prevent the harmful consequences of activation.

Abbreviations: βKO, p38β knock-out, I/R%, myocardial infarction as a percentage of myocardium at risk of infarction, DRα, drug-resistant p38α, DRβ, drug-resistant p38β, HSP27, heat stress protein of 27kDa, MAPK, mitogen-activated protein kinase, MAPKK, mitogen-activated protein kinase kinase, MAPKKK, mitogen-activated protein kinase kinase kinase, P38, p38 mitogen-activated protein kinase, SAPK, stress activated protein kinase, TAB1, transforming growth factorβ-activated protein kinase 1-binding protein 1, T106M, substitution of threonine 106 for methionine, WT, wild type

Keywords: Myocardial infarction, SB203580, Ischemic preconditioning, p38 MAPK, Chemical genetics