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Docosahexaenoic Acid Induced Apoptosis In H9c2 Cells And Changed Cardiac Function After Ischemia-Reperfusion Injury Open Access


Other title
H9c2 cells
Ischemia/reperfusion injury
Type of item
Degree grantor
University of Alberta
Author or creator
Qadhi, Rawabi
Supervisor and department
Seubert, John (Pharmaceutical Sciences)
Examining committee member and department
El-Kadi, Ayman (Pharmaceutical Sciences)
Lavasanifar, Afsaneh (Pharmaceutical Sciences)
Davidge, Sandra (Obstetrics/Gynecology and Physiology)
Faculty of Pharmacy and Pharmaceutical Sciences
Pharmaceutical Sciences
Date accepted
Graduation date
Master of Science
Degree level
Cardiovascular disease (CVD) remains one of the leading causes of death worldwide. As such, a vast amount of research has investigated novel therapies for preventing and/or reducing CVD. Much evidence has demonstrated the importance of dietary composition in increasing or lowering risks of CVD. While the role played by dietary n-3 polyunsaturated fatty acids (PUFAs) in reducing CVD has been recognized for many years, the protective mechanisms of these molecules, notably toward ischemia-reperfusion (IR) injury, remain unknown. Ecosapentaenoic acid (20:5n-3) and docosahexaenoic acid (22:6n-3, DHA) are two of the most important n-3 PUFAs. Both are present in fish oil. The objective of this study is to investigate the effect of acute administration of DHA on IR injury. Methods: Hearts from male C57BL6 mice were isolated and perfused in Langendorff mode and then subjected to IR injury. Hearts were perfused with different concentrations of DHA (0, 10, 50 and 100 µM) to determine its effect on cardiac function and recovery. Mechanistic studies were performed using rat myoblast cells (H9c2 cells) in an anoxia-reoxygenation protocol. Cell viability (MTT assay), cytochrome c release, and caspase-3 and caspase-8 activities were measured to compare cellular injury in DHA treated cells versus controls. The impact of DHA on mitochondrial morphology and function was assessed using epifluorescent microscopy. Results: Data demonstrated that DHA has adverse effects on both pre- and post-ischemic left ventricular developed pressures and on the heart rate, systolic and diastolic heart rates. Cell experiments revealed that significant cell death occurs in a concentration-dependent manner when H9c2 cells are treated with DHA and subjected to anoxia-reoxygenation injury. Moreover, apoptotic cell death is caused by DHA treatment, during which cytochrome c is released and caspases-8 and -3 are activated. Significant mitochondrial fragmentation and loss of membrane potential were observed with high concentrations of DHA. Conclusion: Our data suggest that acute treatment with DHA impedes the function of isolated hearts and triggers apoptotic cell death.
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