3,4-Methylenedioxymethamphetamine activates nuclear factor-kappaB, increases intracellular calcium, and modulates gene transcription in rat heart cells.
3,4-Methylenedioxymethamphetamine (MDMA) is an illicit psychoactive drug that has gained immense popularity among teenagers and young adults. The cardiovascular toxicological consequences of abusing this compound have not been fully characterized. The present study utilized a transient transfection/dual luciferase genetic reporter assay, fluorescence confocal microscopy, and gene expression macroarray technology to determine nuclear factor-kappaB (NF-kappaB) activity, intracellular calcium balance, mitochondrial depolarization, and gene transcription profiles, respectively, in cultured rat striated cardiac myocytes (H9c2) exposed to MDMA. At concentrations of 1 x 10(-3) M and 1 x 10(-2) M, MDMA significantly enhanced NF-kappaB reporter activity compared with 0 M (medium only) control. This response was mitigated by cotransfection with IkappaB for 1 x 10(-3) M but not 1 x 10(-2) M MDMA. MDMA significantly increased intracellular calcium at concentrations of 1 x 10(-3) M and 1 x 10(-2) M and caused mitochondrial depolarization at 1 x 10(-2) M. MDMA increased the transcription of genes that are considered to be biomarkers in cardiovascular disease and genes that respond to toxic insults. Selected gene activation was verified via temperature-gradient RT-PCR conducted with annealing temperatures ranging from 50 degrees C to 65 degrees C. Collectively, these results suggest that MDMA may be toxic to the heart through its ability to activate the myocardial NF-kappaB response, disrupt cytosolic calcium and mitochondrial homeostasis, and alter gene transcription.
Tiangco
David A
DA
Department of Biological Sciences, Old Dominion University, Norfolk, VA 23529, USA.
Lattanzio
Frank A
FA
Osgood
Christopher J
CJ
Beebe
Stephen J
SJ
Kerry
Julie A
JA
Hargrave
Barbara Y
BY
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002478:Cells, Cultured; D015870:Gene Expression / Q000187:drug effects*; D017930:Genes, Reporter / Q000187:drug effects / Q000235:genetics; D006213:Hallucinogens / Q000494:pharmacology*; D008156:Luciferases / Q000378:metabolism; D018613:Microscopy, Confocal; D008929:Mitochondria, Heart / Q000187:drug effects / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000378:metabolism*; D018817:N-Methyl-3,4-methylenedioxyamphetamine / Q000494:pharmacology*; D016328:NF-kappa B / Q000096:biosynthesis* / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction
D006213:Hallucinogens; D016328:NF-kappa B; D008156:Luciferases; D018817:N-Methyl-3,4-methylenedioxyamphetamine; D002118:Calcium