title: Activation of mitochondrial calpain and increased cardiac injury: beyond AIF release.
abstract: Calpain 1 (CPN1) is a ubiquitous cysteine protease that exists in both cytosol and cardiac mitochondria. Mitochondrial CPN1 (mit-CPN1) is located in the intermembrane space and matrix. Activation of mit-CPN1 within the intermembrane space increases cardiac injury by releasing apoptosis-inducing factor from mitochondria during ischemia-reperfusion (IR). We asked if activation of mit-CPN1 is involved in mitochondrial injury during IR. MDL-28170 (MDL) was used to inhibit CPN1 in buffer-perfused hearts following 25-min ischemia and 30-min reperfusion. MDL treatment decreased the release of lactate dehydrogenase into coronary effluent compared with untreated hearts, indicating that inhibition of CPN1 decreases cardiac injury. MDL also prevented the cleavage of spectrin (a substrate of CPN1) in cytosol during IR, supporting that MDL treatment decreased cytosolic calpain activation. In addition, MDL markedly improved calcium retention capacity compared with untreated heart, suggesting that MDL treatment decreases mitochondrial permeability transition pore opening. In addition, we found that IR led to decreased complex I activity, whereas inhibition of mit-CPN1 using MDL protected complex I. Pyruvate dehydrogenase content was decreased following IR. However, pyruvate dehydrogenase content was preserved in MDL-treated mitochondria. Taken together, MDL treatment decreased cardiac injury during IR by inhibiting both cytosolic and mit-CPN1. Activation of mit-CPN1 increases cardiac injury during IR by sensitizing mitochondrial permeability transition pore opening and impairing mitochondrial metabolism through damage of complex I.
authors.affiliation: Division of Cardiology, Department of Medicine, Virginia Commonwealth University, Richmond, Virginia;
authors.forename: Jeremy
authors.initials: J
authors.lastname: Thompson
authors.affiliation: Division of Cardiology, Department of Medicine, Virginia Commonwealth University, Richmond, Virginia;
authors.forename: Ying
authors.initials: Y
authors.lastname: Hu
authors.affiliation: Division of Cardiology, Department of Medicine, Virginia Commonwealth University, Richmond, Virginia; Department of Biochemistry and Molecular Biology, Virginia Commonwealth University, Richmond, Virginia; Department of Physiology and Biophysics, Virginia Commonwealth University, Richmond, Virginia; and McGuire Veterans Affairs Medical Center, Richmond, Virginia.
authors.forename: Edward J
authors.initials: EJ
authors.lastname: Lesnefsky
authors.affiliation: Division of Cardiology, Department of Medicine, Virginia Commonwealth University, Richmond, Virginia; Qun.chen@vcuhealth.org.
authors.forename: Qun
authors.identifier: http://orcid.org/0000-0001-5683-7844
authors.initials: Q
authors.lastname: Chen
mesh_terms: D000818:Animals; D051033:Apoptosis Inducing Factor / Q000187:drug effects* / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002154:Calpain / Q000037:antagonists & inhibitors* / Q000378:metabolism; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology*; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D004151:Dipeptides / Q000494:pharmacology*; D042967:Electron Transport Complex I / Q000187:drug effects / Q000378:metabolism; D006321:Heart / Q000187:drug effects*; D007770:L-Lactate Dehydrogenase / Q000187:drug effects / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008929:Mitochondria, Heart / Q000187:drug effects* / Q000378:metabolism; D033681:Mitochondrial Membrane Transport Proteins / Q000187:drug effects* / Q000378:metabolism; D000083162:Mitochondrial Permeability Transition Pore; D015428:Myocardial Reperfusion Injury / Q000378:metabolism*; D013049:Spectrin / Q000187:drug effects / Q000378:metabolism
chemical_list: D051033:Apoptosis Inducing Factor; D015853:Cysteine Proteinase Inhibitors; D004151:Dipeptides; D033681:Mitochondrial Membrane Transport Proteins; D000083162:Mitochondrial Permeability Transition Pore; C494830:AIFM1 protein, mouse; D013049:Spectrin; D007770:L-Lactate Dehydrogenase; D002154:Calpain; C112682:Capn1 protein, mouse; D042967:Electron Transport Complex I; D002118:Calcium; C058076:calpain inhibitor III
keywords: MPTP; PDH; calcium; complex I; mitochondria