Methamphetamine induces AP-1 and NF-kappaB binding and transactivation in human brain endothelial cells

J Neurosci Res. 2001 Nov 15;66(4):583-91. doi: 10.1002/jnr.1248.

Abstract

Cellular and molecular mechanisms of methamphetamine (METH)-induced neurotoxicity may involve alterations of cellular redox status and induction of inflammatory genes in endothelial cells. To study these hypotheses, molecular signaling pathways of METH-induced inflammatory responses via activation of redox-sensitive transcription factors were investigated in human brain microvascular endothelial cells (HBMEC). A dose-dependent depletion of total glutathione levels was detected in HBMEC exposed to METH. In addition, electrophoretic mobility shift assay (EMSA) showed significant increases in DNA binding activities of redox-responsive transcription factors, AP-1 and NF-kappaB, in HBMEC treated with METH. METH-mediated AP-1 or NF-kappaB activation was accompanied by induction of transactivation of AP-1 or NF-kappaB, as measured by dual luciferase assay using specific reporter plasmids. Because NF-kappaB and AP-1 are known to regulate expression of inflammatory genes, expression of the gene encoding for tumor necrosis factor-alpha (TNF-alpha) was also studied in METH-treated HBMEC. A dose-dependent overexpression of the TNF-alpha gene was observed in HBMEC treated with METH. The importance of AP-1 and NF-kappaB in METH-induced TNF-alpha gene was confirmed in functional promoter studies using constructs of the TNF-alpha promoter with mutated AP-1 or NF-kappaB sites. These results indicate that METH-induced disturbances in cellular redox status and activation of AP-1 and NF-kappaB can play critical roles in the signaling pathways leading to upregulation of inflammatory genes in human brain endothelial cells.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amphetamine-Related Disorders / genetics
  • Amphetamine-Related Disorders / metabolism
  • Amphetamine-Related Disorders / physiopathology
  • Binding Sites / drug effects
  • Binding Sites / physiology
  • Blood-Brain Barrier / drug effects
  • Blood-Brain Barrier / physiology
  • Brain / drug effects*
  • Brain / metabolism
  • Brain / physiopathology
  • Cells, Cultured / drug effects
  • Cells, Cultured / metabolism
  • Encephalitis / chemically induced
  • Encephalitis / genetics*
  • Encephalitis / metabolism
  • Endothelium, Vascular / drug effects*
  • Endothelium, Vascular / metabolism
  • Endothelium, Vascular / physiopathology
  • Gene Expression Regulation / drug effects
  • Gene Expression Regulation / physiology
  • Genes, Reporter / drug effects
  • Genes, Reporter / physiology
  • Glutathione / drug effects
  • Glutathione / metabolism
  • Humans
  • Methamphetamine / toxicity*
  • Microcirculation / drug effects
  • Microcirculation / metabolism
  • Microcirculation / physiopathology
  • NF-kappa B / drug effects*
  • NF-kappa B / genetics
  • NF-kappa B / metabolism
  • Oxidation-Reduction / drug effects
  • Oxidative Stress / drug effects*
  • Oxidative Stress / genetics
  • RNA, Messenger / drug effects
  • RNA, Messenger / metabolism
  • Signal Transduction / drug effects
  • Signal Transduction / genetics
  • Transcription Factor AP-1 / drug effects*
  • Transcription Factor AP-1 / genetics
  • Transcription Factor AP-1 / metabolism
  • Transcription, Genetic / drug effects
  • Transcription, Genetic / physiology
  • Transfection
  • Tumor Necrosis Factor-alpha / drug effects
  • Tumor Necrosis Factor-alpha / genetics
  • Tumor Necrosis Factor-alpha / metabolism

Substances

  • NF-kappa B
  • RNA, Messenger
  • Transcription Factor AP-1
  • Tumor Necrosis Factor-alpha
  • Methamphetamine
  • Glutathione