Endogenous morphine signaling via nitric oxide regulates the expression of CYP2D6 and COMT: autocrine/paracrine feedback inhibition.
We determined changes in mRNA expression in specific enzymes involved in the biosynthesis of morphine in human white blood cells via microarray. Leukocyte exposure to morphine down-regulated catechol-O-methyl transferase (COMT) and CYP2D6 by approximately 50% compared with control values. The treatment did not alter DOPA decarboxylase and dopamine beta-hydroxylase expression, demonstrating the specificity of morphine actions. The verification of the microarray data was accomplished via real-time Taqman reverse transcriptase polymerase chain reaction (RT-PCR) focused on CYP2D6 and COMT expression in different blood samples treated with morphine. The analysis showed similar changes in the expression of CYP2D6 and COMT mRNA. The expression was reduced by 47 +/- 7% for CYP2D6, substantiating the microarray finding of a 54% reduction. Furthermore, exposure of white blood cells to 10(-6) M S-nitroso-N-acetyl-DL-penicillamine (SNAP), a nitric oxide (NO) donor, reduced the expression of CYP2D6 and COMT. Prior naloxone (10(-6) M) or N-nitro-L-arginine methyl ester (L-NAME) (10(-4) M) addition abrogated morphine's down-regulating activity, demonstrating morphine was initiating its actions via stimulating constitutive NO synthase derived NO release via the mu3 opiate receptor splice variant. In the past we demonstrated that UDP-glucurosyltransferase is involved in metabolizing morphine to morphine 6-glucuronide in adrenal chromaffin cells. In the present study its expression was not found in controls and morphine-treated cells, suggesting that morphine 6-glucuronide may not be synthesized in white blood cells. Taken together, it appears that morphine has the ability to modulate its own synthesis via autocrine and paracrine signaling.
Mantione
Kirk J
KJ
Neuroscience Research Institute, State University of New York-College at Old Westbury, 11568, USA.
Cadet
Patrick
P
Zhu
Wei
W
Kream
Richard M
RM
Sheehan
Melinda
M
Fricchione
Gregory L
GL
Goumon
Yannick
Y
Esch
Tobias
T
Stefano
George B
GB
D019898:Autocrine Communication / Q000235:genetics*; D002394:Catechol O-Methyltransferase / Q000235:genetics*; D019389:Cytochrome P-450 CYP2D6 / Q000235:genetics*; D015536:Down-Regulation / Q000235:genetics; D025461:Feedback, Physiological / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D006801:Humans; D007962:Leukocytes / Q000187:drug effects / Q000378:metabolism; D009020:Morphine / Q000378:metabolism* / Q000494:pharmacology*; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009270:Naloxone / Q000494:pharmacology; D009292:Narcotic Antagonists / Q000494:pharmacology; D009569:Nitric Oxide / Q000502:physiology*; D020411:Oligonucleotide Array Sequence Analysis; D019899:Paracrine Communication / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics*; D017450:Receptors, Opioid, mu / Q000187:drug effects / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D026423:S-Nitroso-N-Acetylpenicillamine / Q000494:pharmacology; D015398:Signal Transduction / Q000235:genetics*
D009292:Narcotic Antagonists; D012333:RNA, Messenger; D017450:Receptors, Opioid, mu; D009569:Nitric Oxide; D009270:Naloxone; D009020:Morphine; D026423:S-Nitroso-N-Acetylpenicillamine; D019389:Cytochrome P-450 CYP2D6; D002394:Catechol O-Methyltransferase; D019331:NG-Nitroarginine Methyl Ester