Huiliang Zhang1, Guohua Gong2, Pei Wang2, Zhen Zhang2, Stephen C Kolwicz2, Peter S Rabinovitch3, Rong Tian2, Wang Wang4. 1. Mitochondria and Metabolism Center, Department of Anesthesiology and Pain Medicine, University of Washington, Seattle, WA 98109, USA; Department of Pathology, University of Washington, Seattle, WA, 98195, USA. 2. Mitochondria and Metabolism Center, Department of Anesthesiology and Pain Medicine, University of Washington, Seattle, WA 98109, USA. 3. Department of Pathology, University of Washington, Seattle, WA, 98195, USA. 4. Mitochondria and Metabolism Center, Department of Anesthesiology and Pain Medicine, University of Washington, Seattle, WA 98109, USA; Department of Pathology, University of Washington, Seattle, WA, 98195, USA. Electronic address: wangwang@uw.edu.
Abstract
RATIONALE: Ischemic heart disease (IHD) is a leading cause of mortality. The most effective intervention for IHD is reperfusion, which ironically causes ischemia reperfusion (I/R) injury mainly due to oxidative stress-induced cardiomyocyte death. The exact mechanism and site of reactive oxygen species (ROS) generation during I/R injury remain elusive. OBJECTIVE: We aim to test the hypothesis that Complex I-mediated forward and reverse electron flows are the major source of ROS in I/R injury of the heart. METHODS AND RESULTS: We used a genetic model of mitochondrial Complex I deficiency, in which a Complex I assembling subunit, Ndufs4 was knocked out in the heart (Ndufs4H-/-). The Langendorff perfused Ndufs4H-/- hearts exhibited significantly reduced infarct size (45.3 ± 5.5% in wild type vs 20.9 ± 8.1% in Ndufs4H-/-), recovered contractile function, and maintained mitochondrial membrane potential after no flow ischemia and subsequent reperfusion. In cultured adult cardiomyocytes from Ndufs4H-/- mice, I/R mimetic treatments caused minimal cell death. Reintroducing Ndufs4 in Ndufs4H-/- cardiomyocytes abolished the protection. Mitochondrial NADH declined much slower in Ndufs4H-/- cardiomyocytes during reperfusion suggesting decreased forward electron flow. Mitochondrial flashes, a marker for mitochondrial respiration, were inhibited in Ndufs4H-/- cardiomyocytes at baseline and during I/R, which was accompanied by preserved aconitase activity suggesting lack of oxidative damage. Finally, pharmacological blockade of forward and reverse electron flow at Complex I inhibited I/R-induced cell death. CONCLUSIONS: These results provide the first genetic evidence supporting the central role of mitochondrial Complex I in I/R injury of mouse heart. The study also suggests that both forward and reverse electron flows underlie oxidative cardiomyocyte death during reperfusion.
RATIONALE: Ischemic heart disease (IHD) is a leading cause of mortality. The most effective intervention for IHD is reperfusion, which ironically causes ischemia reperfusion (I/R) injury mainly due to oxidative stress-induced cardiomyocyte death. The exact mechanism and site of reactive oxygen species (ROS) generation during I/R injury remain elusive. OBJECTIVE: We aim to test the hypothesis that Complex I-mediated forward and reverse electron flows are the major source of ROS in I/R injury of the heart. METHODS AND RESULTS: We used a genetic model of mitochondrial Complex I deficiency, in which a Complex I assembling subunit, Ndufs4 was knocked out in the heart (Ndufs4H-/-). The Langendorff perfused Ndufs4H-/- hearts exhibited significantly reduced infarct size (45.3 ± 5.5% in wild type vs 20.9 ± 8.1% in Ndufs4H-/-), recovered contractile function, and maintained mitochondrial membrane potential after no flow ischemia and subsequent reperfusion. In cultured adult cardiomyocytes from Ndufs4H-/- mice, I/R mimetic treatments caused minimal cell death. Reintroducing Ndufs4 in Ndufs4H-/- cardiomyocytes abolished the protection. Mitochondrial NADH declined much slower in Ndufs4H-/- cardiomyocytes during reperfusion suggesting decreased forward electron flow. Mitochondrial flashes, a marker for mitochondrial respiration, were inhibited in Ndufs4H-/- cardiomyocytes at baseline and during I/R, which was accompanied by preserved aconitase activity suggesting lack of oxidative damage. Finally, pharmacological blockade of forward and reverse electron flow at Complex I inhibited I/R-induced cell death. CONCLUSIONS: These results provide the first genetic evidence supporting the central role of mitochondrial Complex I in I/R injury of mouse heart. The study also suggests that both forward and reverse electron flows underlie oxidative cardiomyocyte death during reperfusion.
Authors: Karol Szczepanek; Qun Chen; Marta Derecka; Fadi N Salloum; Qifang Zhang; Magdalena Szelag; Joanna Cichy; Rakesh C Kukreja; Jozef Dulak; Edward J Lesnefsky; Andrew C Larner Journal: J Biol Chem Date: 2011-06-29 Impact factor: 5.157
Authors: Mohammed Aldakkak; David F Stowe; Qun Chen; Edward J Lesnefsky; Amadou K S Camara Journal: Cardiovasc Res Date: 2008-01-15 Impact factor: 10.787
Authors: Aijun Xu; Karol Szczepanek; Michael W Maceyka; Thomas Ross; Elizabeth Bowler; Ying Hu; Barrett Kenny; Chris Mehfoud; Pooja N Desai; Clive M Baumgarten; Qun Chen; Edward J Lesnefsky Journal: Am J Physiol Cell Physiol Date: 2014-04-02 Impact factor: 4.249
Authors: Guangyu Zhang; Xiaoding Wang; Chao Li; Qinfeng Li; Yu A An; Xiang Luo; Yingfeng Deng; Thomas G Gillette; Philipp E Scherer; Zhao V Wang Journal: Circulation Date: 2021-09-29 Impact factor: 29.690
Authors: Leonardo Maciel; Dahienne Ferreira de Oliveira; Gustavo Monnerat; Antonio Carlos Campos de Carvalho; Jose Hamilton Matheus Nascimento Journal: Front Pharmacol Date: 2020-05-05 Impact factor: 5.810
Authors: Orli Thau-Zuchman; Linda Svendsen; Simon C Dyall; Ursula Paredes-Esquivel; Molly Rhodes; John V Priestley; René G Feichtinger; Barbara Kofler; Susanne Lotstra; J Martin Verkuyl; Robert J Hageman; Laus M Broersen; Nick van Wijk; Jose P Silva; Jordi L Tremoleda; Adina T Michael-Titus Journal: Theranostics Date: 2021-01-01 Impact factor: 11.556
Authors: Adam R Wende; John C Schell; Chae-Myeong Ha; Mark E Pepin; Oleh Khalimonchuk; Hansjörg Schwertz; Renata O Pereira; Manoja K Brahma; Joseph Tuinei; Ariel Contreras-Ferrat; Li Wang; Chase A Andrizzi; Curtis D Olsen; Wayne E Bradley; Louis J Dell'Italia; Wolfgang H Dillmann; Sheldon E Litwin; E Dale Abel Journal: Diabetes Date: 2020-05-04 Impact factor: 9.461
Authors: Huiliang Zhang; Nathan N Alder; Wang Wang; Hazel Szeto; David J Marcinek; Peter S Rabinovitch Journal: Elife Date: 2020-12-15 Impact factor: 8.140