Pretreatment with insulin before ischaemia reduces infarct size in Langendorff-perfused rat hearts.
OBJECTIVE
To compare the possible role of Akt and mammalian target of rapamycin (mTOR) in mediating cardioprotection against ischaemia under three different conditions: (1) During ischaemic preconditioning (IPC), (2) when insulin was given as a pretreatment agent (InsPC) and (3) when insulin was given as a reperfusion cell survival agent (Ins(R)).


METHODS
Isolated perfused rat hearts were subjected to IPC (3 x 5 min) or InsPC (50 mU mL(-1); 3 x 5 min), before 30 min of regional ischaemia followed by 120 min of reperfusion +/- 1L-6-hydroxymethyl-chiro-inositol-2-[(R)-2-O-methyl-3-O-octadecylcarbonate] (HIMO) (20 microm; Akt inhibitor) or rapamycin (1 nm; mTOR inhibitor). In addition, insulin (3 mU mL(-1)) was given at the onset of reperfusion, +/- HIMO or rapamycin. Risk zone (R) and infarct size (I) were determined with Evans blue and tetrazolium staining respectively. Western blot analysis was performed on tissue from Langendorff-perfused rat hearts and cell lysates from cultured HL1 cells.


RESULTS
IPC, InsPC and InsR treatment resulted in a significant reduction in infarct size compared to controls (all P < 0.05). This protective effect of IPC and insulin was abolished by the inhibitors. However, the putative Akt inhibitor, although capable of abolishing cardioprotection induced by insulin, was not able to inhibit insulin-induced phosphorylation of Akt in Langendorff-perfused rat hearts and cultured HL1 cells. The target for this compound therefore remains to be determined.


CONCLUSIONS
IPC and insulin (either as InsPC or Ins(R)) appear to activate mTOR, and this kinase seems to play an essential role in cardioprotection against ischaemia and reperfusion injury as rapamycin blocked the protection.
Fuglesteg
B N
BN
Department of Medical Physiology, Faculty of Medicine, University of Tromsø, Tromsø, Norway. brittf@fagmed.uit.no
Tiron
C
C
Jonassen
A K
AK
Mjøs
O D
OD
Ytrehus
K
K
D000818:Animals; D007004:Hypoglycemic Agents / Q000627:therapeutic use*; D007328:Insulin / Q000627:therapeutic use*; D019157:Ischemic Preconditioning, Myocardial / Q000379:methods*; D008297:Male; D023421:Models, Animal; D009203:Myocardial Infarction / Q000517:prevention & control*; D051058:Oncogene Protein v-akt / Q000037:antagonists & inhibitors; D010477:Perfusion; D011494:Protein Kinases / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D020123:Sirolimus / Q000627:therapeutic use; D058570:TOR Serine-Threonine Kinases
D007004:Hypoglycemic Agents; D007328:Insulin; D011494:Protein Kinases; C546845:mTOR protein, rat; D051058:Oncogene Protein v-akt; D058570:TOR Serine-Threonine Kinases; D020123:Sirolimus