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Myocardial Ischemia and Preconditioning pp 205–218Cite as

Role of Mitochondrial Membrane Potential in Cardiac Protection against Ischemia

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Part of the book series: Progress in Experimental Cardiology ((PREC,volume 6))

Summary

Our previous study has indicated that diazoxide protected myocardium against ischemia-reperfusion injury. This study tests the hypothesis that the maintenance of mitochondrial membrane potential (ΔΨm) in myocytes is responsible for cell protection against ischemia. This was specifically tested in myocytes after activation of the mitoKATP channel. Myocyte damage by 3 hrs anoxia and 2 hrs reoxygenation (A-R) was evaluated by cell viability, membrane permeability and apoptosis. Mitochondrial function was indicated by the concentration of ATP. Mitochondrial morphology was observed by staining myocytes with Mito Tracker Orange CMTMRos and by electron microscopy. Immunostaining was used to determine the distribution of cytochrome c. ΔΨm was assayed by staining with 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolcarbocyanine iodide (JC-1) and observed by con-focal microscopy. Results show that 1) An extensive damage was observed in cultured myocytes as evidenced by decreased cell viability, compromised membrane permeability, increased apoptosis and decreased ATP concentration after A-R. 2) Mitochondria in A-R myocytes were swollen and exhibited a collapsed ΔΨm. Cytochrome c was released from mitochondria into the cytosol. 3) Diazoxide (100μmol/L) significantly prevented myocyte and mitochondrial damage, cytochrome c loss, and stabilized ΔΨm. 4) This protection was blocked by 5-hydroxydecanoate (5-HD, 500μmol/L), a mitoKATP channel selective inhibitor but not by HMR-1098 (30μmol/L), a putative sarcolemmal KATP channel selective inhibitor. 5) Diazoxide reduced ΔΨm in normal cultured myocytes in a concentration- and time-independent pattern. It is concluded that activation of mitoKATP channel with diazoxide prevented disruption of ΔΨm resulting in protection against A-R induced injury.

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Authors and Affiliations

  1. Department of Pathology and Laboratory Medicine, University of Cincinnati, 231 Albert Sabin Way, 45267-0529, Cincinnati, OH, USA

    Meifeng Xu, Yigang Wang, Ahmar Ayub & Muhammad Ashraf PhD

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  1. Meifeng Xu
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  2. Yigang Wang
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  4. Muhammad Ashraf PhD
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Corresponding author

Correspondence to Muhammad Ashraf PhD .

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Editors and Affiliations

  1. Institute of Cardiovascular Sciences St. Boniface General Hospital Research Centre Faculty of Medicine, University of Manitoba, Winnipeg, Canada

    Naranjan S. Dhalla PhD, MD (Hon), DSc (Hon) (Distinguished Professor and Director) & Anton Lukas PhD (Associate Professor) (Distinguished Professor and Director) &  (Associate Professor)

  2. Aoto Hospital, Department of Internal Medicine, Jikei University School of Medicine, Tokyo, Japan

    Nobuakira Takeda MD, PhD (Associate Professor) (Associate Professor)

  3. Dept. of Pharmaceutical Sciences & Drug Research, Punjabi University, Patiala, India

    Manjeet Singh PhD (Professor & Head) (Professor & Head)

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Xu, M., Wang, Y., Ayub, A., Ashraf, M. (2003). Role of Mitochondrial Membrane Potential in Cardiac Protection against Ischemia. In: Dhalla, N.S., Takeda, N., Singh, M., Lukas, A. (eds) Myocardial Ischemia and Preconditioning. Progress in Experimental Cardiology, vol 6. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-0355-2_15

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  • DOI: https://doi.org/10.1007/978-1-4615-0355-2_15

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4613-5036-1

  • Online ISBN: 978-1-4615-0355-2

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