Gene and Protein Expression Profiles of Anti‐ and Pro‐apoptotic Actions of Dopamine, R‐Apomorphine, Green Tea Polyphenol (−)‐Epigallocatechine‐3‐gallate, and Melatonin
Identifieur interne : 001765 ( Main/Exploration ); précédent : 001764; suivant : 001766Gene and Protein Expression Profiles of Anti‐ and Pro‐apoptotic Actions of Dopamine, R‐Apomorphine, Green Tea Polyphenol (−)‐Epigallocatechine‐3‐gallate, and Melatonin
Auteurs : Orly Weinreb [Israël] ; Silvia Mandel [Israël] ; Moussa B. H. Youdim [Israël]Source :
- Annals of the New York Academy of SciencesNEUROPROTECTIVE AGENTS: Sixth International Conference [ 0077-8923 ] ; 2003-05.
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Abstract
Abstract: Significant evidence has been provided to support the hypothesis that oxidant stress may be responsible for degeneration of dopaminergic neurons in the substantia nigra pars compacta in Parkinson's disease. Dopamine (DA), R‐apomorphine (R‐APO), green tea polyphenol (−)‐epigallocatechine‐3‐gallate (EGCG), and melatonin are neuroprotective and radical scavenger compounds. The aim of this study was to establish the mechanism of the concentration‐dependent neuroprotective and pro‐apoptotic action of these drugs via gene expression and protein determination. cDNA microarrays provide new prospects to study and identify various mechanisms of drug action. We employed this technique for the study reported in this paper. Total RNA was extracted from SH‐SY5Y cells exposed to low neuroprotective and high toxic concentrations of the drugs, followed by synthesis of cDNA, and hybridization to a microarray membrane related to apoptosis, survival, and cell cycle pathways. We demonstrated a concentration and time‐dependent correlation between R‐APO, DA, EGCG, and melatonin in modulation of cell survival/cell death‐related gene pathways. The results were confirmed by quantitative real‐time PCR and protein profiles. Unlike the effects of low concentrations (1‐10 μM), where an antiapoptotic response was manifest, a proapoptotic pattern of gene expression was observed at high toxic concentrations (50‐500 μM) of the antioxidants (e.g., increase in caspases, fas, and gadd45). Our results have provided novel insights into the gene mechanisms involved in both the neuroprotective and proapoptotic activities of neuroprotective drugs. We have shown that DA, R‐APO, EGCG, and melatonin exhibit similar gene expression and protein profiles.
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DOI: 10.1111/j.1749-6632.2003.tb07544.x
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<front><div type="abstract" xml:lang="en">Abstract: Significant evidence has been provided to support the hypothesis that oxidant stress may be responsible for degeneration of dopaminergic neurons in the substantia nigra pars compacta in Parkinson's disease. Dopamine (DA), R‐apomorphine (R‐APO), green tea polyphenol (−)‐epigallocatechine‐3‐gallate (EGCG), and melatonin are neuroprotective and radical scavenger compounds. The aim of this study was to establish the mechanism of the concentration‐dependent neuroprotective and pro‐apoptotic action of these drugs via gene expression and protein determination. cDNA microarrays provide new prospects to study and identify various mechanisms of drug action. We employed this technique for the study reported in this paper. Total RNA was extracted from SH‐SY5Y cells exposed to low neuroprotective and high toxic concentrations of the drugs, followed by synthesis of cDNA, and hybridization to a microarray membrane related to apoptosis, survival, and cell cycle pathways. We demonstrated a concentration and time‐dependent correlation between R‐APO, DA, EGCG, and melatonin in modulation of cell survival/cell death‐related gene pathways. The results were confirmed by quantitative real‐time PCR and protein profiles. Unlike the effects of low concentrations (1‐10 μM), where an antiapoptotic response was manifest, a proapoptotic pattern of gene expression was observed at high toxic concentrations (50‐500 μM) of the antioxidants (e.g., increase in caspases, fas, and gadd45). Our results have provided novel insights into the gene mechanisms involved in both the neuroprotective and proapoptotic activities of neuroprotective drugs. We have shown that DA, R‐APO, EGCG, and melatonin exhibit similar gene expression and protein profiles.</div>
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