Abstract
Ischemia–reperfusion syndromes of the heart and brain are the leading cause of death and long-term disability worldwide. Development of effective treatments for myocardial infarction, stroke, cardiac arrest and their sequelae requires preclinical models that replicate specific features of ischemia–reperfusion. The complexities of intact animals, including the integrated function of organ systems, autonomic innervation and endocrine factors, often preclude detailed study of specific components of ischemia–reperfusion injury cascades. Ischemia represents the interruption of metabolic fuel and oxygen delivery to support cellular oxidative metabolism; reintroduction of oxygen upon reperfusion of ischemic tissue triggers oxidative stress which initiates the reperfusion injury cascade culminating in injury and death of cells and tissues. Thus, cultured cells subjected to hypoxia, fuel deprivation and reoxygenation replicate the cardinal features of ischemia–reperfusion, while accommodating interventions such as siRNA suppression of specific genes and pharmacological activation or inhibition of signaling cascades that are not feasible in more complex preparations, especially intact animals. This chapter describes an in vitro OGD-reoxygenation cell culture model, an excellent preparation to examine the cellular mechanisms mediating ischemia–reperfusion injury and/or cytoprotection.
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References
Ryou MG, Choudhury GR, Li W, Winters A, Yuan F, Liu R, Yang SH (2015) Methylene blue-induced neuronal protective mechanism against hypoxia-reoxygenation stress. Neuroscience 301:193–203
Frank A, Bonney M, Bonney S, Weitzel L, Koeppen M, Eckle T (2012) Myocardial ischemia reperfusion injury: from basic science to clinical bedside. Semin Cardiothorac Vasc Anesth 16(3):123–132
Green AR (2008) Pharmacological approaches to acute ischaemic stroke: reperfusion certainly, neuroprotection possibly. Br J Pharmacol 153(Suppl 1):S325–S338
Xie L, Li W, Winters A, Yuan F, Jin K, Yang S (2013) Methylene blue induces macroautophagy through 5′ adenosine monophosphate-activated protein kinase pathway to protect neurons from serum deprivation. Front Cell Neurosci 7:56
Choi JR, Pingguan-Murphy B, Wan Abas WA, Yong KW, Poon CT, Noor Azmi MA, Omar SZ, Chua KH, Xu F, Wan Safwani WK (2015) In situ normoxia enhances survival and proliferation rate of human adipose tissue-derived stromal cells without increasing the risk of tumourigenesis. PLoS One 10(1):e0115034
Yang C, Jiang L, Zhang H, Shimoda LA, DeBerardinis RJ, Semenza GL (2014) Analysis of hypoxia-induced metabolic reprogramming. Methods Enzymol 542:425–455
Acknowledgments
This work was supported by UNTHSC intramural research grant RI6148 and grant NS076975 from National Institute of Neurological Disorders and Stroke.
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Ryou, Mg., Mallet, R.T. (2018). An In Vitro Oxygen–Glucose Deprivation Model for Studying Ischemia–Reperfusion Injury of Neuronal Cells. In: Tharakan, B. (eds) Traumatic and Ischemic Injury. Methods in Molecular Biology, vol 1717. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7526-6_18
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DOI: https://doi.org/10.1007/978-1-4939-7526-6_18
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-7526-6
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