Neuroprotective effect of fraxetin and myricetin against rotenone-induced apoptosis in neuroblastoma cells.

Abstract

Rotenone-induced apoptosis is considered to contribute to the etiology of Parkinson's disease (PD). We try to prevent the apoptosis induced by rotenone toxicity with 50 microM myricetin, 100 microM fraxetin and 100 microM N-acetylcysteine (NAC) that protect against reactive oxygen species (ROS), on SH-SY5Y human neuroblastoma cell line. Morphological changes induced by rotenone and intracellular ROS were assessed in live SH-SY5Y dopaminergic cells by confocal microscopy using the fluorescent dyes, dihydroethidium and 2',7'-dichlorofluorescein diacetate (DCFH-DA). DNA fragmentation was assayed as index of apoptosis. We also investigated oxidative stress parameters such as the glutathione redox status and lipid peroxidation. The exposure of the SH-SY5Y cells to rotenone 5 microM for 16 h produced severe morphological changes, DNA fragmentation and significative increases in the levels of hydrogen peroxide and superoxide anion. These increases were reduced by a 30-min pretreatment with fraxetin 100 microM or NAC 100 microM. DNA laddering produced by rotenone treatment was also inhibited by fraxetin and NAC. Treatment with 5 microM rotenone induced loss of reduced glutathione (GSH) and increased cellular levels of oxidized glutathione (GSSG). Fraxetin and NAC treatments restored glutathione redox ratio diminished after rotenone challenge and decreased the levels of lipid peroxidation. These results suggest that the natural antioxidants, such as fraxetin, may prevent the apoptotic death of dopaminergic cells induced by rotenone and mediated by oxidative stress.

• Authors: María Francisca Molina-Jiménez, María Isabel Sánchez-Reus, David Andres, María Cascales, Juana Benedi
• Journal: Brain research (Brain Res) Vol. 1009 Issue 1-2 Pg. 9-16 (May 29 2004) ISSN: 0006-8993 [Print] Netherlands
• PMID: 15120578 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Coumarins
• Flavonoids
• Fluoresceins
• Neuroprotective Agents
• Reactive Oxygen Species
• Rotenone
• dihydroethidium
• rotenolone
• diacetyldichlorofluorescein
• myricetin
• DNA
• fraxetin
• Ethidium
• Glutathione
• Acetylcysteine

Topics

• Acetylcysteine (pharmacology)
• Analysis of Variance
• Apoptosis
• Blotting, Southern (methods)
• Cell Line, Tumor
• Cell Survival (drug effects)
• Coumarins (pharmacology)
• DNA (metabolism)
• Dose-Response Relationship, Drug
• Ethidium (analogs & derivatives, metabolism)
• Flavonoids (pharmacology)
• Fluoresceins (metabolism)
• Glutathione (metabolism)
• Humans
• Lipid Peroxidation (drug effects)
• Microscopy, Confocal (methods)
• Neuroblastoma (pathology)
• Neuroprotective Agents (pharmacology)
• Reactive Oxygen Species (metabolism)
• Rotenone (analogs & derivatives, antagonists & inhibitors)
• Time Factors