title: Partial inhibition of the ubiquitin-proteasome system ameliorates cardiac dysfunction following ischemia-reperfusion in the presence of high glucose.
abstract: BACKGROUND
  Acute hyperglycemia co-presenting with myocardial infarction (in diabetic and non-diabetic individuals) is often associated with a poor prognosis. Although acute hyperglycemia induces oxidative stress that can lead to dysregulation of the ubiquitin-proteasome system (UPS), it is unclear whether increased/decreased UPS is detrimental with ischemia-reperfusion under such conditions. As our earlier data implicated the UPS in cardiac damage, we hypothesized that its inhibition results in cardioprotection with ischemia-reperfusion performed under conditions that simulate acute hyperglycemia.
  
  
  METHODS
  Ex vivo rat heart perfusions were performed with Krebs-Henseleit buffer containing 33 mM glucose vs. controls (11 mM glucose) for 60 min stabilization, followed by 20 min global ischemia and 60 min reperfusion ± 5 µM lactacystin and 10 µM MG-132, respectively. The UPS inhibitors were added during the first 20 min of the reperfusion phase and various cardiac functional parameters evaluated. In parallel experiments, infarct sizes were assessed following 20 min regional ischemia and 120 min reperfusion ± each of the respective UPS inhibitors (added during reperfusion). Heart tissues were collected and analyzed for markers of oxidative stress, UPS activation, inflammation and autophagy.
  
  
  RESULTS
  The proteasome inhibitor doses and treatment duration here employed resulted in partial UPS inhibition during the reperfusion phase. Both lactacystin and MG-132 administration resulted in cardioprotection in our experimental system, with MG-132 showing a greater effect. The proteasome inhibitors also enhanced cardiac superoxide dismutase protein levels (SOD1, SOD2), attenuated pro-inflammatory effects and caused an upregulation of autophagic markers.
  
  
  CONCLUSIONS
  This study established that partial proteasome inhibition elicits cardioprotection in hearts exposed to ischemia-reperfusion with acute simulated hyperglycemia. These data reveal that protease inhibition triggered three major protective effects, i.e. (a) enhancing myocardial anti-oxidant defenses, (b) attenuating inflammation, and (c) increasing the autophagic response. Thus the UPS emerges as a unique therapeutic target for the treatment of ischemic heart disease under such conditions.
authors.affiliation: Cardio-Metabolic Research Group (CMRG), Department of Physiological Sciences, Stellenbosch University, Room 2005, Mike De Vries Building, Merriman Avenue, Stellenbosch, 7600, South Africa. adamsbuin@gmail.com.
authors.forename: Buin
authors.initials: B
authors.lastname: Adams
authors.affiliation: Cardio-Metabolic Research Group (CMRG), Department of Physiological Sciences, Stellenbosch University, Room 2005, Mike De Vries Building, Merriman Avenue, Stellenbosch, 7600, South Africa. mapangarudo@sun.ac.za.
authors.forename: Rudo F
authors.initials: RF
authors.lastname: Mapanga
authors.affiliation: Cardio-Metabolic Research Group (CMRG), Department of Physiological Sciences, Stellenbosch University, Room 2005, Mike De Vries Building, Merriman Avenue, Stellenbosch, 7600, South Africa. mfessop@sun.ac.za.
authors.forename: M Faadiel
authors.initials: MF
authors.lastname: Essop
mesh_terms: D000111:Acetylcysteine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology; D000975:Antioxidants / Q000494:pharmacology; D001343:Autophagy / Q000187:drug effects; D058065:Diabetic Cardiomyopathies / Q000201:enzymology / Q000473:pathology / Q000503:physiopathology / Q000517:prevention & control*; D006943:Hyperglycemia / Q000188:drug therapy* / Q000201:enzymology; D018836:Inflammation Mediators / Q000378:metabolism; D000067268:Isolated Heart Preparation; D007976:Leupeptins / Q000494:pharmacology*; D008297:Male; D015428:Myocardial Reperfusion Injury / Q000201:enzymology / Q000473:pathology / Q000503:physiopathology / Q000517:prevention & control*; D009206:Myocardium / Q000201:enzymology* / Q000473:pathology; D061988:Proteasome Inhibitors / Q000494:pharmacology*; D017208:Rats, Wistar; D013482:Superoxide Dismutase / Q000378:metabolism; D000072105:Superoxide Dismutase-1; D013997:Time Factors; D000099156:Superoxide Dismutase 2
chemical_list: D000111:Acetylcysteine; D000893:Anti-Inflammatory Agents; D000975:Antioxidants; D018836:Inflammation Mediators; D007976:Leupeptins; D061988:Proteasome Inhibitors; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1; D000099156:Superoxide Dismutase 2; C067713:lactacystin; C000606292:Sod1 protein, rat; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde