Attenuation of myocardial ischaemic injury 24 h after diacylglycerol treatment in vivo.
Previous work suggests that delayed protection against infarction following ischaemic preconditioning of rabbit myocardium may involve the activation of protein kinase C (PKC). Preconditioning in the presence of chelerythrine, an inhibitor of PKC, abolished the late anti-infarct effect of preconditioning. In the studies described here, we tested the hypothesis that direct PKC activation with an analogue of diacylglycerol, the physiological activator of PKC, would invoke an adaptive mechanism leading to enhanced myocardial tolerance to ischaemia 24 h later. Rabbits were treated with i.v. injections of 1,2-dioctanoyl-sn-glycerol (DiC8) 5 mg/kg or 15 mg/kg or an equivalent volume of vehicle solution. Twenty-four h after pretreatment, the animals were anaesthetised and underwent 30 min coronary artery occlusion with 120 min reperfusion. Infarct size was determined by triphenyltetrazolium staining. In vehicle pretreated rabbits, infarct-risk zone ratio was 32.8+/-2.6%. Pretreatment with DiC8 5 mg/kg did not significantly affect infarct size (26.3+/-4.0%), but pretreatment with DiC8 15 mg/kg resulted in a marked reduction in infarct size (18.0+/-3.4%, P<0.05, 1-way ANOVA). Reduction in infarct size with the higher dose of DiC8 was independent of myocardial risk zone size and systemic haemodynamic parameters during coronary occlusion. The haemodynamic effects of acute administration of DiC8 15 mg/kg were examined in a separate cohort of pentobarbitone-anaesthetised rabbits. The compound was found not to affect systolic blood pressure or heart rate under these conditions. We examined the possibility that increased ischaemic tolerance might be due to induction of the 27 and 72 kDa heat shock proteins (hsp27 and hsp70i) which are known to be cytoprotective and are upregulated by ischaemia and other stressful stimuli. Western blot analysis of left ventricular tissue revealed that neither protein was induced 24 h after treatment with DiC8 15 mg/kg. Thus, infarct limitation 24 h after DiC8 treatment did not appear to be due to increased tissue content of these proteins. The mechanisms of DiC8-induced delayed myocardial protection remain unclear. However, these data are compatible with the hypothesis that activation of PKC isoenzymes is an important intermediate signal of sub-acute cellular adaptation, and results in enhanced tolerance to ischaemia-reperfusion injury in myocardium many hours later.
Baxter
G F
GF
The Hatter Institute for Cardiovascular Studies, University College London Hospital and Medical School, London, UK.
Mocanu
M M
MM
Yellon
D M
DM
D000818:Animals; D001794:Blood Pressure; D004075:Diglycerides / Q000494:pharmacology*; D004789:Enzyme Activation; D006339:Heart Rate; D006360:Heat-Shock Proteins / Q000378:metabolism; D006439:Hemodynamics; D006863:Hydrogen-Ion Concentration; D008297:Male; D009203:Myocardial Infarction / Q000473:pathology; D017202:Myocardial Ischemia / Q000188:drug therapy*; D011493:Protein Kinase C / Q000378:metabolism*; D011817:Rabbits; D015427:Reperfusion Injury / Q000378:metabolism; D013997:Time Factors
D004075:Diglycerides; D006360:Heat-Shock Proteins; C044295:1,2-dioctanoylglycerol; D011493:Protein Kinase C