Effect of fraxetin and myricetin on rotenone-induced cytotoxicity in SH-SY5Y cells: comparison with N-acetylcysteine.

Abstract

The purpose of this study was to investigate the potential neuroprotective effects of myricetin (flavonoid) and fraxetin (coumarin) on rotenone-induced apoptosis in SH-SY5Y cells, and the possible signal pathway involved in a neuronal cell model of Parkinson's disease. These two compounds were compared to N-acetylcysteine. The viability of cells was assessed by 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and cytotoxicity was assayed by lactate dehydrogenase (LDH) released into the culture medium. Parameters related to apoptosis, such as caspase-3 activity, the cleavage of poly(ADP-ribose) polymerase and the levels of reactive oxygen species were also determined. Rotenone caused a time- and dose-dependent decrease in cell viability and the degree of LDH release was proportionally to the effects on cell viability. Cells were pretreated with fraxetin, myricetin and N-acetylcysteine at different concentrations for 30 min before exposure to rotenone. Cytotoxicity of rotenone (5 microM) for 16 h was significantly diminished as well as the release of LDH into the medium, by the effect of fraxetin, myricetin and N-acetylcysteine, with fraxetin (100 microM) and N-acetylcysteine (100 microM) being more effective than myricetin (50 microM). Rotenone-induced apoptosis in SH-SY5Y cells was detected by an increase in caspase-3 activity and in the cleavage of poly(ADP-ribose) polymerase. After exposing these cells to rotenone, a significant increase in reactive oxygen species preceded apoptotic events. Fraxetin (100 microM) and N-acetylcysteine (100 microM) not only reduced rotenone-induced reactive oxygen species formation, but also attenuated caspase-3 activity and poly(ADP-ribose) polymerase cleavage at 16 h against rotenone-induced apoptosis. The effect of fraxetin in both experiments was similar to that of N-acetylcysteine. These results demonstrated the protective action of fraxetin and suggest that it can reduce apoptosis, possibly by decreasing free radical generation in SH-SY5Y cells. Myricetin at 100 microM was without any preventive effect.

• Authors: M Francisca Molina-Jimenez, M Isabel Sanchez-Reus, Juana Benedi
• Journal: European journal of pharmacology (Eur J Pharmacol) Vol. 472 Issue 1-2 Pg. 81-7 (Jul 04 2003) ISSN: 0014-2999 [Print] Netherlands
• PMID: 12860476 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Coumarins
• Flavonoids
• Neuroprotective Agents
• Reactive Oxygen Species
• Rotenone
• myricetin
• fraxetin
• L-Lactate Dehydrogenase
• CASP3 protein, human
• Caspase 3
• Caspases
• Acetylcysteine

Topics

• Acetylcysteine (pharmacology)
• Apoptosis (drug effects)
• Caspase 3
• Caspases (metabolism)
• Cell Line, Tumor
• Cell Survival (drug effects)
• Coumarins (pharmacology)
• Flavonoids (pharmacology)
• Humans
• L-Lactate Dehydrogenase (metabolism)
• Neuroblastoma (enzymology, pathology)
• Neuroprotective Agents (pharmacology)
• Reactive Oxygen Species (metabolism)
• Rotenone (toxicity)