title: Damage to mitochondrial complex I during cardiac ischemia reperfusion injury is reduced indirectly by anti-anginal drug ranolazine.
abstract: Ranolazine, an anti-anginal drug, is a late Na(+) channel current blocker that is also believed to attenuate fatty acid oxidation and mitochondrial respiratory complex I activity, especially during ischemia. In this study, we investigated if ranolazine's protective effect against cardiac ischemia/reperfusion (IR) injury is mediated at the mitochondrial level and specifically if respiratory complex I (NADH Ubiquinone oxidoreductase) function is protected. We treated isolated and perfused guinea pig hearts with ranolazine just before 30 min ischemia and then isolated cardiac mitochondria at the end of 30 min ischemia and/or 30 min ischemia followed by 10 min reperfusion. We utilized spectrophotometric and histochemical techniques to assay complex I activity, Western blot analysis for complex I subunit NDUFA9, electron paramagnetic resonance for activity of complex I Fe-S clusters, enzyme linked immuno sorbent assay (ELISA) for determination of protein acetylation, native gel histochemical staining for respiratory supercomplex assemblies, and high pressure liquid chromatography for cardiolipin integrity; cardiac function was measured during IR. Ranolazine treated hearts showed higher complex I activity and greater detectable complex I protein levels compared to untreated IR hearts. Ranolazine treatment also led to more normalized electron transfer via Fe-S centers, supercomplex assembly and cardiolipin integrity. These improvements in complex I structure and function with ranolazine were associated with improved cardiac function after IR. However, these protective effects of ranolazine are not mediated by a direct action on mitochondria, but rather indirectly via cytosolic mechanisms that lead to less oxidation and better structural integrity of complex I.
authors.affiliation: Department of Anesthesiology, Medical College of Wisconsin, Milwaukee, WI 53226, USA.
authors.forename: Ashish K
authors.initials: AK
authors.lastname: Gadicherla
authors.forename: David F
authors.initials: DF
authors.lastname: Stowe
authors.forename: William E
authors.initials: WE
authors.lastname: Antholine
authors.forename: Meiying
authors.initials: M
authors.lastname: Yang
authors.forename: Amadou K S
authors.initials: AK
authors.lastname: Camara
mesh_terms: D000083:Acetanilides / Q000627:therapeutic use*; D000818:Animals; D015153:Blotting, Western; D002308:Cardiolipins / Q000032:analysis / Q000502:physiology; D004579:Electron Transport; D042967:Electron Transport Complex I / Q000032:analysis / Q000502:physiology*; D006168:Guinea Pigs; D008929:Mitochondria, Heart / Q000187:drug effects / Q000378:metabolism*; D015428:Myocardial Reperfusion Injury / Q000378:metabolism / Q000517:prevention & control*; D010879:Piperazines / Q000627:therapeutic use*; D000069458:Ranolazine; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D000083:Acetanilides; D002308:Cardiolipins; D010879:Piperazines; D017382:Reactive Oxygen Species; D000069458:Ranolazine; D042967:Electron Transport Complex I