Neuroprotective effects of orientin on hydrogen peroxide-induced apoptosis in SH-SY5Y cells

Neurodegenerative diseases remain a global issue which affects the ageing population. Efforts towards determining their aetiologies to understand their pathogenic mechanisms are underway in order to identify a pathway through which therapeutic measures can be applied. One such pathogenic mechanism,...

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Published in:	Molecular medicine reports Vol. 9; no. 3; pp. 947 - 954
Main Authors:	LAW, BENJAMIN NGEE TIING, LING, ANNA PICK KIONG, KOH, RHUN YIAN, CHYE, SOI MOI, WONG, YING PEI
Format:	Journal Article
Language:	English
Published:	Greece D.A. Spandidos 01-03-2014 Spandidos Publications Spandidos Publications UK Ltd
Subjects:	Analysis Antioxidants Apoptosis Apoptosis - drug effects Blood-brain barrier Care and treatment Caspase Caspase 3 - metabolism Caspase 7 - metabolism Caspase 8 - metabolism Caspase 9 - metabolism Caspase-9 Cell cycle Cell Cycle Checkpoints - drug effects Cell Line, Tumor Control Cytochrome Cytotoxicity Dementia Disease Ethanol Flavonoids Flavonoids - pharmacology Flow cytometry Glucosides - pharmacology Health aspects Humans Hydrogen peroxide Hydrogen Peroxide - toxicity Intracellular Isoflavones Mortality Nervous system diseases Neuroblastoma cells Neurodegenerative diseases Neuroprotection Neuroprotective Agents - pharmacology Older people orientin Oxidative stress Reactive oxygen species Reactive Oxygen Species - metabolism Studies United States > US
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Summary:	Neurodegenerative diseases remain a global issue which affects the ageing population. Efforts towards determining their aetiologies to understand their pathogenic mechanisms are underway in order to identify a pathway through which therapeutic measures can be applied. One such pathogenic mechanism, oxidative stress (OS), is widely considered to be involved in neurodegenerative disease. Antioxidants, most notably flavonoids, have promising potential for therapeutic use as shown in in vitro and in vivo studies. In view of the importance of flavonoids for combating OS, this study investigated the neuroprotective effects of orientin, which has been reported to be capable of crossing the blood-brain barrier. The maximum non-toxic dose (MNTD) of orientin against SH-SY5Y neuroblastoma cells was determined using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The effects of the MNTD and the half MNTD (½MNTD) of orientin on cell cycle progression and intracellular reactive oxygen species (ROS) levels, as well as the activity of caspases 3/7, 8 and 9 after exposure to 150 μM of hydrogen peroxide (H2O2) were also determined using flow cytometry, a 2′,7′-dichlorodihydrofluorescein-diacetate (DCFH-DA) assay and caspase assay kits, respectively. The results revealed that orientin at ≤20 μM was not cytotoxic to SH-SY5Y cells. After treatment with orientin at the MNTD, the percentage of apoptotic cells was significantly reduced compared with that in cells treated with 150 μM H2O2 alone. The results also showed that, although orientin at the MNTD and ½MNTD did not reduce intracellular ROS levels, it significantly inhibited the activity of caspases 3/7. Caspase 9 was significantly inactivated with orientin at the MNTD. Findings from this study suggest that the neuroprotection conferred by orientin was the result of the intracellular mediation of caspase activity.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	1791-2997 1791-3004
DOI:	10.3892/mmr.2013.1878