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Alpha linolenic acid decreases apoptosis and oxidized phospholipids in cardiomyocytes during ischemia/reperfusion

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Abstract

The omega-3 fatty acid, alpha linolenic acid (ALA) found in plant-derived foods induces significant cardiovascular benefits when ingested. ALA may be cardioprotective during ischemia; however, the mechanism(s) responsible for this effect is unknown. Isolated adult rat cardiomyocytes were exposed to medium containing ALA for 24 h and then exposed to non-ischemic (control), simulated ischemia (ISCH), or simulated ischemia/reperfusion (IR) conditions. Cardiomyocyte phospholipids were extracted and analyzed by an HPLC/electrospray ionization tandem mass spectrometry system. Pre-treatment of cells with ALA resulted in a significant incorporation of ALA within cardiomyocyte phosphatidylcholine. Cell death, DNA fragmentation and caspase-3 activity increased during ischemia and ischemia/reperfusion. Two pro-apoptotic oxidized phosphatidylcholine (OxPC) species, 1-palmitoyl-2-(5′-oxo-valeroyl)-sn-glycero-3-phosphocholine (POVPC), and 1-palmitoyl-2-glutaroyl-sn-glycero-3-phosphocholine (PGPC) were significantly increased during both ischemia and ischemia/reperfusion. Pre-treatment of the cells with ALA resulted in a significant reduction in cell death during ischemia and ischemia/reperfusion challenge. Apoptosis was also inhibited during ischemia and ischemia/reperfusion as shown by reduced DNA fragmentation and decreased caspase activation. ALA pre-treatment significantly decreased the production of POVPC and PGPC during ischemia and ischemia/reperfusion. ALA pre-treatment also significantly increased in resting Ca2+ during ischemia or ischemia/reperfusion but did not improve Ca2+ transients. ALA protects the cardiomyocyte from apoptotic cell death during simulated ISCH and IR by inhibiting the production of specific pro-apoptotic OxPC species. OxPCs represent a viable interventional target to protect the heart during ischemic challenge.

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Abbreviations

ALA:

α-Linolenic acid

CL:

Cardiolipin

ESI:

Electrospray ionization

HPLC:

High performance liquid chromatography

ISCH:

Ischemia

IR:

Ischemia reperfusion

KDdiA- PPC:

1-palmitoyl-2-(4-keto-dodec-3-enadioyl)-sn-glycero-3-phosphocholine

KOdiA-PPC:

1-palmitoyl-2-(5-keto-6-octene-dioyl)-sn-glycero-3-phosphocholine

LDL:

Low-density lipoprotein

MRM:

Multiple reaction monitoring

nadph:

Nicotinamide adenine dinucleotide phosphate

OxLDL:

Oxidized low-density lipoprotein

OxPAPC:

Oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine

OxPC:

Oxidized phosphatidylcholine

OxPL:

Oxidized phospholipids

PAzPC:

1-palmitoyl-2-azelaoyl-sn-glycero-3-phosphocholine

PC:

Phosphatidylcholine

PE:

Phosphatidylethanolamine

PEIPC:

1-palmitoyl-2-(5,6-epoxyisoprostane E2)-sn-glycero-3-phosphocholine

PGPC:

1-palmitoyl-2-glutaryl-sn-glycero-3-phosphocholine

PI:

Phosphatidylinositol

PL:

Phospholipid

PPARγ:

Peroxisome proliferator-activator receptor gamma

PONPC:

1-palmitoyl-2-(5′-oxo-valeroyl)-sn-glycero-3-phosphocholine

POVPC:

1-palmitoyl-2-(9′-oxo-nonanoyl)-sn-glycero-3-phosphocholine

PS:

Phosphatidylserine

PSPC:

1-palmitoyl-2-stearoyl-sn-glycero-3-phosphocholine

PUFA:

Poly-unsaturated fatty acid

ROS:

Reactive oxygen species

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Acknowledgements

This study was supported by operating grants obtained from the Canadian Institutes for Health Research (to GNP) and from Manitoba Medical Services Foundation (to AR) and the Heart and Stroke Foundation of Canada (to AR). Indirect support for this research was obtained from St Boniface Hospital Research Foundation. RG was supported by a Studentship from the Manitoba Health Research Council. DH was supported by a joint-funded Studentship from the Manitoba Health Research Council and the University of Manitoba.

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Authors and Affiliations

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

    Riya Ganguly, Devin Hasanally, Thane G. Maddaford, Rakesh Chaudhary, Grant N. Pierce & Amir Ravandi

  2. Canadian Centre for Agri-food Research in Health and Medicine (CCARM), St. Boniface Hospital Research Centre, University of Manitoba, Winnipeg, MB, Canada

    Riya Ganguly, Thane G. Maddaford & Grant N. Pierce

  3. Departments of Physiology and Pathophysiology, Faculty of Medicine, St. Boniface Hospital Research Centre, University of Manitoba, Winnipeg, MB, Canada

    Riya Ganguly, Devin Hasanally, Aleksandra Stamenkovic, Thane G. Maddaford, Grant N. Pierce & Amir Ravandi

  4. Section of Cardiology, Faculty of Medicine, St. Boniface Hospital Research Centre, University of Manitoba, Winnipeg, MB, Canada

    Amir Ravandi

  5. Albrechtsen Research Centre, St. Boniface Hospital, 351 Taché Avenue, Winnipeg, MB, R2H 2A6, Canada

    Grant N. Pierce & Amir Ravandi

Authors
  1. Riya Ganguly
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  2. Devin Hasanally
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  3. Aleksandra Stamenkovic
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  4. Thane G. Maddaford
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  5. Rakesh Chaudhary
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  6. Grant N. Pierce
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  7. Amir Ravandi
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Contributions

Riya Ganguly: Collection and analysis of data, writing of manuscript; Devin Hasanally: Collection and analysis of data, writing of paper; Aleksandra Stamenkovic: Collection and analysis of data. Thane G Maddaford: Collection and analysis of data, writing of paper; Rakesh Chaudhary: Collection and analysis of data; Grant N Pierce: Coordination of experiments, writing of paper; Amir Ravandi: Coordination of experiments, writing of paper.

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Correspondence to Grant N. Pierce or Amir Ravandi.

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Co-first authorship: Riya Ganguly and Devin Hasanally.

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Ganguly, R., Hasanally, D., Stamenkovic, A. et al. Alpha linolenic acid decreases apoptosis and oxidized phospholipids in cardiomyocytes during ischemia/reperfusion. Mol Cell Biochem 437, 163–175 (2018). https://doi.org/10.1007/s11010-017-3104-z

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