GENERATION OF SURVIVAL SIGNAL BY DIFFERENTIAL INTERACTION OF p38MAPKα and p38MAPKβ WITH CAVEOLIN-1 AND CAVEOLIN-3 IN THE ADAPTED HEART

摘要

Sphingomyelin breakdown product ceramide has recently been found to induce an adaptive response and reduce myocardial ischemia/reperfusion injury. Since activation of MAP kinases plays an essential role in myocardial adaptation to ischemic stress and since ceramide is involved in lipid raft formation where MAP kinases can be translocated in response to stress, we reasoned that preconditioning may potentiate the translocation of MAP kinases into the lipid raft. To test the hypothesis, rats were divided into five groups: i) control, ii) ischemia/reperfusion (I/R), iii) I/R + C2-ceramide, iv) adapted and v) adapted+ desipramine, an inhibitor of ceramide formation. Isolated hearts were preperfused for 15 min with Krebs Henseleit Bicarbonate (KHB) buffer in the absence or presence of 10 μM desipramine followed by adaptation induced by four cyclic episodes of 5 min ischemia and 10 min reperfusion. For myocardial adaptation to ischemia with ceramide, the hearts were perfused with 1μM C-2-ceramide. All hearts were then subjected to 30 min ischemia and 2 h of reperfusion. As expected, both ischemic adaptation and ceramide adaptation made the heart resistant to I/R injury as evidenced by improved ventricular performance and reduced myocardial infarct size and cardiomyocyte apoptosis, which were significantly blocked with desipramine indicating the involvement of ceramide in ischemic adaptation. Ceramide also participated in the formation of lipid raft, and desipramine disrupted the raft formation. In the adapted hearts, there was an increased association of the proapoptotic p38MAPKα with caveolin-1 while there was a reduced association of anti-apoptotic p38MAPKβ with caveolin-3 indicating reduced amount of p38MAPKα and increased amount of p38MAPKβ were available to the adapted hearts thereby generating a survival signal. Desipramine decreased the association of P38MAPKα and C-2 ceramide increased the association of P38MAPKα with the lipid raft. The survival signal was further confirmed by increased phosphorylation of AKT and enhanced induction of expression of Bcl-2 during adaptation and its reversal with desipramine. The results indicated a unique ceramide signaling the ischemic and PC hearts involving lipid rafts, which generated a survival signal by differentially associating the p38MAPKα and p38MAPKβ with the caveolin-1 and caveoli-3, respectively.