Ascorbate Protects Neurons against Oxidative Stress: A Raman Microspectroscopic Study

Dutta, Abhaya and Gautam, Rekha and Chatterjee, Sreejata and Ariese, Freek and Sikdar, Sujit Kumar and Umapathy, Siva (2015) Ascorbate Protects Neurons against Oxidative Stress: A Raman Microspectroscopic Study. In: ACS CHEMICAL NEUROSCIENCE, 6 (11). pp. 1794-1801.

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Abstract

Oxidative stress due to excessive accumulation of reactive oxygen or nitrogen species in the brain as seen in certain neurodegenerative diseases can have deleterious effects on neurons. Hydrogen peroxide, endogenously generated in neurons under normal physiological conditions, can produce an excess of hydroxyl radical via a Fenton mediated mechanism. This may induce acute oxidative injury if not scavenged or removed effectively by antioxidants. There are several biochemical assay methods to estimate oxidative injury in cells; however, they do not provide information on the biochemical changes as the cells get damaged progressively under oxidative stress. Raman microspectroscopy offers the possibility of real time monitoring of the chemical composition of live cells undergoing oxidative stress under physiological conditions. In the present study, a hippocampal neuron coculture was used to observe the acute impact of hydroxyl radicals generated by hydrogen peroxide in the presence of Fe2+ (Fenton reaction). Raman peaks related to nucleic acids (725, 782, 1092, 1320, 1340, 1420, and 1576 cm(-1)) showed time-dependent changes over the experimental period (60 mm), indicating the breakdown of the phosphodiester backbone as well as nuclear bases. Interestingly, ascorbic acid (a potent antioxidant) when cotreated with Fenton reactants showed protection of cells as inferred from the Raman spectra, presumably by scavenging hydroxyl radicals. Little or no change in the Raman spectra was observed for untreated control cells and for cells exposed to Fe2+ only, H2O2 only, and ascorbate only. A live dead assay study also supported the current observations. Hence, Raman microspectroscopy has the potential to be an excellent noninvasive tool for early detection of oxidative stress that is seen in neurodegenerative diseases.

Item Type:	Journal Article
Publication:	ACS CHEMICAL NEUROSCIENCE
Publisher:	AMER CHEMICAL SOC
Additional Information:	Copy right for this article belongs to the AMER CHEMICAL SOC, 1155 16TH ST, NW, WASHINGTON, DC 20036 USA
Keywords:	Raman spectroscopy; hippocampal neuron; oxidative stress; neurodegenerative disease; reactive oxygen species
Department/Centre:	Division of Biological Sciences > Molecular Biophysics Unit Division of Chemical Sciences > Inorganic & Physical Chemistry Division of Physical & Mathematical Sciences > Instrumentation Appiled Physics
Date Deposited:	15 Dec 2015 07:47
Last Modified:	15 Dec 2015 07:47
URI:	http://eprints.iisc.ac.in/id/eprint/52914

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