EGCG Protects against 6-OHDA-Induced Neurotoxicity in a Cell Culture Model

Background. Parkinson’s disease (PD) is a progressive neurodegenerative disease that causes severe brain dopamine depletion. Disruption of iron metabolism may be involved in the PD progression. Objective. To test the protective effect of (−)-epigallocatechin-3-gallate (EGCG) against 6-hydroxydopamin...

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Main Authors: Dan Chen, Anumantha G. Kanthasamy, Manju B. Reddy
Format: Article
Language:English
Published: Wiley 2015-01-01
Series:Parkinson's Disease
Online Access:http://dx.doi.org/10.1155/2015/843906
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author Dan Chen
Anumantha G. Kanthasamy
Manju B. Reddy
author_facet Dan Chen
Anumantha G. Kanthasamy
Manju B. Reddy
author_sort Dan Chen
collection DOAJ
description Background. Parkinson’s disease (PD) is a progressive neurodegenerative disease that causes severe brain dopamine depletion. Disruption of iron metabolism may be involved in the PD progression. Objective. To test the protective effect of (−)-epigallocatechin-3-gallate (EGCG) against 6-hydroxydopamine- (6-OHDA-) induced neurotoxicity by regulating iron metabolism in N27 cells. Methods. Protection by EGCG in N27 cells was assessed by SYTOX green assay, MTT, and caspase-3 activity. Iron regulatory gene and protein expression were measured by RT-PCR and Western blotting. Intracellular iron uptake was measured using 55Fe. The EGCG protection was further tested in primary mesencephalic dopaminergic neurons by immunocytochemistry. Results. EGCG protected against 6-OHDA-induced cell toxicity. 6-OHDA treatment significantly (p<0.05) increased divalent metal transporter-1 (DMT1) and hepcidin and decreased ferroportin 1 (Fpn1) level, whereas pretreatment with EGCG counteracted the effects. The increased 55Fe (by 96%, p<0.01) cell uptake confirmed the iron burden by 6-OHDA and was reduced by EGCG by 27% (p<0.05), supporting the DMT1 results. Pretreatment with EGCG and 6-OHDA significantly increased (p<0.0001) TH+ cell count (~3-fold) and neurite length (~12-fold) compared to 6-OHDA alone in primary mesencephalic neurons. Conclusions. Pretreatment with EGCG protected against 6-OHDA-induced neurotoxicity by regulating genes and proteins involved in brain iron homeostasis, especially modulating hepcidin levels.
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spelling doaj-art-240d970c07064988a1cc9dfe2ba649c42025-08-20T02:04:40ZengWileyParkinson's Disease2090-80832042-00802015-01-01201510.1155/2015/843906843906EGCG Protects against 6-OHDA-Induced Neurotoxicity in a Cell Culture ModelDan Chen0Anumantha G. Kanthasamy1Manju B. Reddy2Department of Food Sciences and Human Nutrition, Iowa State University, 220 Mackay Hall, Ames, IA 50010, USADepartment of Biomedical Sciences, Iowa State University, Ames, IA 50010, USADepartment of Food Sciences and Human Nutrition, Iowa State University, 220 Mackay Hall, Ames, IA 50010, USABackground. Parkinson’s disease (PD) is a progressive neurodegenerative disease that causes severe brain dopamine depletion. Disruption of iron metabolism may be involved in the PD progression. Objective. To test the protective effect of (−)-epigallocatechin-3-gallate (EGCG) against 6-hydroxydopamine- (6-OHDA-) induced neurotoxicity by regulating iron metabolism in N27 cells. Methods. Protection by EGCG in N27 cells was assessed by SYTOX green assay, MTT, and caspase-3 activity. Iron regulatory gene and protein expression were measured by RT-PCR and Western blotting. Intracellular iron uptake was measured using 55Fe. The EGCG protection was further tested in primary mesencephalic dopaminergic neurons by immunocytochemistry. Results. EGCG protected against 6-OHDA-induced cell toxicity. 6-OHDA treatment significantly (p<0.05) increased divalent metal transporter-1 (DMT1) and hepcidin and decreased ferroportin 1 (Fpn1) level, whereas pretreatment with EGCG counteracted the effects. The increased 55Fe (by 96%, p<0.01) cell uptake confirmed the iron burden by 6-OHDA and was reduced by EGCG by 27% (p<0.05), supporting the DMT1 results. Pretreatment with EGCG and 6-OHDA significantly increased (p<0.0001) TH+ cell count (~3-fold) and neurite length (~12-fold) compared to 6-OHDA alone in primary mesencephalic neurons. Conclusions. Pretreatment with EGCG protected against 6-OHDA-induced neurotoxicity by regulating genes and proteins involved in brain iron homeostasis, especially modulating hepcidin levels.http://dx.doi.org/10.1155/2015/843906
spellingShingle Dan Chen
Anumantha G. Kanthasamy
Manju B. Reddy
EGCG Protects against 6-OHDA-Induced Neurotoxicity in a Cell Culture Model
Parkinson's Disease
title EGCG Protects against 6-OHDA-Induced Neurotoxicity in a Cell Culture Model
title_full EGCG Protects against 6-OHDA-Induced Neurotoxicity in a Cell Culture Model
title_fullStr EGCG Protects against 6-OHDA-Induced Neurotoxicity in a Cell Culture Model
title_full_unstemmed EGCG Protects against 6-OHDA-Induced Neurotoxicity in a Cell Culture Model
title_short EGCG Protects against 6-OHDA-Induced Neurotoxicity in a Cell Culture Model
title_sort egcg protects against 6 ohda induced neurotoxicity in a cell culture model
url http://dx.doi.org/10.1155/2015/843906
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AT manjubreddy egcgprotectsagainst6ohdainducedneurotoxicityinacellculturemodel