Effect of acute and chronic psychostimulant drugs on redox status, AP-1 activation and pro-enkephalin mRNA in the human astrocyte-like U373 MG cells.
In order to approach the astroglial implication of addictive and neurotoxic processes associated with psychostimulant drug abuse, the effects of amphetamine or cocaine (1-100 microM) on redox status, AP-1 transcription factor and pro-enkephalin, an AP-1 target gene, were investigated in the human astrocyte-like U373 MG cells. We demonstrated an early increase in the generation of radical oxygen species and in the formation of 4-hydroxynonenal-adducts reflecting the pro-oxidant action of both substances. After 1 h or 96 h of treatment, Fos and Jun protein levels were altered and the DNA-binding activity of AP-1 was increased in response to both substances. Using supershift experiments, we observed that the composition of AP-1 dimer differed according to the substance and the duration of treatment. FRA-2 protein represented the main component of the chronic amphetamine- or cocaine-activated complexes, which suggests its relevance in the long-term effects of psychostimulant drugs. Concomitantly, the pro-enkephalin gene was differently regulated by either 6 h or 96 h of treatment. Because astrocytes interact extensively with the neurons in the brain, our data led us to conclude that oxidation-regulated AP-1 target genes may represent one of the molecular mechanisms underlying neuronal adaptation associated with psychostimulant dependence.
Malaplate-Armand
Catherine
C
INSERM U525, Faculté de Pharmacie, Université Henri Poincaré-Nancy I, France. malarmand@free.fr
Becuwe
Philippe
P
Ferrari
Luc
L
Masson
Christine
C
Dauça
Michel
M
Visvikis
Sophie
S
Lambert
Henri
H
Batt
Anne-Marie
AM
D027322:Amino Acid Transport System X-AG / Q000235:genetics / Q000378:metabolism; D000661:Amphetamine / Q000494:pharmacology*; D000704:Analysis of Variance; D001253:Astrocytes / Q000187:drug effects*; D015153:Blotting, Western / Q000379:methods; D002460:Cell Line; D002491:Central Nervous System Agents / Q000494:pharmacology*; D003042:Cocaine / Q000494:pharmacology*; D004268:DNA-Binding Proteins / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004334:Drug Administration Schedule; D024202:Electrophoretic Mobility Shift Assay / Q000379:methods; D004745:Enkephalins / Q000235:genetics / Q000378:metabolism*; D027342:Excitatory Amino Acid Transporter 2 / Q000235:genetics / Q000378:metabolism; D005452:Fluoresceins; D051921:Fos-Related Antigen-2; D005786:Gene Expression Regulation / Q000187:drug effects; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D006801:Humans; D010084:Oxidation-Reduction / Q000187:drug effects; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism*; D016760:Proto-Oncogene Proteins c-fos / Q000378:metabolism; D016755:Proto-Oncogene Proteins c-jun / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013778:Tetrazolium Salts; D013844:Thiazoles; D013997:Time Factors; D018808:Transcription Factor AP-1 / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism; D000099169:Fos-Related Antigen 1
D027322:Amino Acid Transport System X-AG; D000661:Amphetamine; D002491:Central Nervous System Agents; D003042:Cocaine; D004268:DNA-Binding Proteins; D004745:Enkephalins; D027342:Excitatory Amino Acid Transporter 2; D005452:Fluoresceins; D051921:Fos-Related Antigen-2; D005904:Glial Fibrillary Acidic Protein; D011498:Protein Precursors; D016760:Proto-Oncogene Proteins c-fos; D016755:Proto-Oncogene Proteins c-jun; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D013778:Tetrazolium Salts; D013844:Thiazoles; D018808:Transcription Factor AP-1; D014157:Transcription Factors; D000099169:Fos-Related Antigen 1; C497872:FOSL2 protein, human; C029992:proenkephalin; C029569:diacetyldichlorofluorescein; C022616:thiazolyl blue