Increased O-GlcNAc levels during reperfusion lead to improved functional recovery and reduced calpain proteolysis.
We have previously shown that preischemic treatment with glucosamine improved cardiac functional recovery following ischemia-reperfusion, and this was mediated, at least in part, via enhanced flux through the hexosamine biosynthesis pathway and subsequently elevated O-linked N-acetylglucosamine (O-GlcNAc) protein levels. However, preischemic treatment is typically impractical in a clinical setting; therefore, the goal of this study was to investigate whether increasing protein O-GlcNAc levels only during reperfusion also improved recovery. Isolated perfused rat hearts were subjected to 20 min of global, no-flow ischemia followed by 60 min of reperfusion. Administration of glucosamine (10 mM) or an inhibitor of O-GlcNAcase, O-(2-acetamido-2-deoxy-D-glucopyranosylidene)amino-N-phenylcarbamate (PUGNAc; 200 microM), during the first 20 min of reperfusion significantly improved cardiac functional recovery and reduced troponin release during reperfusion compared with untreated control. Both interventions also significantly increased the levels of protein O-GlcNAc and ATP levels. We also found that both glucosamine and PUGNAc attenuated calpain-mediated proteolysis of alpha-fodrin as well as Ca(2+)/calmodulin-dependent protein kinase II during reperfusion. Thus two independent strategies for increasing protein O-GlcNAc levels in the heart during reperfusion significantly improved recovery, and this was correlated with attenuation of calcium-mediated proteolysis. These data provide further support for the concept that increasing cardiac O-GlcNAc levels may be a clinically relevant cardioprotective strategy and suggest that this protection could be due, at least in part, to inhibition of calcium-mediated stress responses.
Liu
Jia
J
Department of Cell Biology, University of Alabama at Birmingham, AL 35294-0005, USA.
Marchase
Richard B
RB
Chatham
John C
JC
D000117:Acetylglucosamine / Q000031:analogs & derivatives / Q000378:metabolism* / Q000627:therapeutic use; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000502:physiology; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism; D002154:Calpain / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism; D005944:Glucosamine / Q000627:therapeutic use; D008840:Microfilament Proteins / Q000378:metabolism; D017202:Myocardial Ischemia / Q000378:metabolism; D015428:Myocardial Reperfusion Injury / Q000378:metabolism* / Q000517:prevention & control*; D009206:Myocardium / Q000378:metabolism; D010091:Oximes / Q000627:therapeutic use; D048448:Phenylcarbamates / Q000627:therapeutic use; D051381:Rats; D013049:Spectrin
D000117:Acetylglucosamine; D000255:Adenosine Triphosphate; D002118:Calcium; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D002154:Calpain; D002352:Carrier Proteins; D005944:Glucosamine; D008840:Microfilament Proteins; D010091:Oximes; D048448:Phenylcarbamates; D013049:Spectrin; C068836:N-acetylglucosaminono-1,5-lactone O-(phenylcarbamoyl)oxime