Ischemic preconditioning protection against stunning in conscious diabetic sheep: role of glucose, insulin, sarcolemmal and mitochondrial KATP channels.
BACKGROUND
Sarcolemmal and mitochondrial ATP-sensitive potassium (KATP) channels have been postulated to participate in preconditioning protection against infarction and stunning. However, these structures appear to be altered in diabetes and thus, it would be possible that preconditioning does not develop in diabetic hearts.


OBJECTIVE
The purpose of this study was to know whether early (EP) and late (LP) ischemic preconditioning against stunning develop in conscious diabetic (D) sheep and whether diabetes affects KATP channel function.


METHODS
Male castrated sheep received alloxan monohydrate (1 g) and were ascribed to three experimental groups: control [DC, 12 min of ischemia (i) followed by 2 h of reperfusion (r)], early preconditioning (DEP, six 5 min i-5 min r periods were performed 45 min before the 12 min i) and late preconditioning (DLP, same as DEP except that the preconditioning stimulus was performed 24 h before the 12 min i). Regional mechanics during reperfusion was evaluated by wall thickening fraction (%WTH) and expressed as percentage of basal values (100%), and KATP channel behavior was indirectly assessed by monophasic action potential duration (MAPD) in relation to its sensitivity to glibenclamide blockade (0.1 and 0.4 mg/kg). The results were compared to those obtained in normal (N) sheep. The effects of sarcolemmal and mitochondrial KATP channel blockade on recovery from stunning were assessed by administration of glibenclamide (0.1 and 0.4 mg/kg) and 5-hydroxydecanoate (5-HD, 5 mg/kg i.v.) and/or diazoxide (10 microg/kg/min over 90 min). Whether acute hyperglycemia (H) in normal animals and insulin (I) treatment in diabetic sheep affected preconditioning protection and KATP channel behavior were also evaluated.


RESULTS
Results expressed as mean % recovery of %WTH showed that preconditioning protected against stunning in normal sheep (NC=65+/-3.5, NLP=82+/-6**, NEP=76+/-4*, *P<0.05 and **P<0.01 against NC) while this did not occur in diabetic ones, where DLP (58+/-7.6) afforded a similar recovery to DC (54+/-5) and DEP worsened instead of improving mechanical function (37+/-9, P<0.01 against DC). Acute hyperglycemia did not affect preconditioning development (NEPH=72+/-3 and NLPH=80+/-4) and insulin treatment reverted the lack of early and late preconditioning protection in diabetic hearts (DEPI=72+/-4* and DLPI=76+/-3*, P<0.05 against DC). Sarcolemmal KATP channel behavior appeared altered in diabetic hearts as shown by MAPD in normal sheep (276+10 ms) compared to diabetic ones (365+9 ms, P<0.05) and by the sensitivity to glibenclamide [0.1 mg/kg completely blocked KATP channels in diabetic (P<0.05) but not in normal hearts]. Insulin also restored MAPD in diabetic heart. Mitochondrial KATP channels appeared not to account for the reported results in diabetes, since glibenclamide (%WTH=40+/-4, P<0.01 vs. NC), but not 5HD nor diazoxide affected myocardial functional recovery during reperfusion.


CONCLUSIONS
Sarcolemmal KATP channel dysfunction due to the lack of insulin affords a primary approach to explain the absence of preconditioning protection against stunning in diabetic sheep hearts.
del Valle
Hector F
HF
Department of Physiology, Pharmacology and Biochemistry, Favaloro University, Buenos Aires, Argentina. delvalle@favaloro.edu.ar
Lascano
Elena C
EC
Negroni
Jorge A
JA
D000200:Action Potentials / Q000187:drug effects; D000704:Analysis of Variance; D000818:Animals; D003652:Decanoic Acids / Q000494:pharmacology; D003921:Diabetes Mellitus, Experimental / Q000378:metabolism*; D003981:Diazoxide / Q000494:pharmacology; D005947:Glucose / Q000502:physiology*; D005905:Glyburide / Q000494:pharmacology; D006880:Hydroxy Acids / Q000494:pharmacology; D007328:Insulin / Q000502:physiology*; D019157:Ischemic Preconditioning, Myocardial / Q000379:methods*; D008297:Male; D008929:Mitochondria, Heart / Q000378:metabolism*; D017682:Myocardial Stunning / Q000517:prevention & control*; D026902:Potassium Channel Blockers / Q000494:pharmacology; D015221:Potassium Channels / Q000502:physiology*; D012508:Sarcolemma / Q000378:metabolism*; D012756:Sheep; D013997:Time Factors
D003652:Decanoic Acids; D006880:Hydroxy Acids; D007328:Insulin; D026902:Potassium Channel Blockers; D015221:Potassium Channels; C052853:5-hydroxydecanoic acid; D005947:Glucose; D003981:Diazoxide; D005905:Glyburide