Morphine-induced nitric oxide production in PC12 cells.
BACKGROUND
The role of nitric oxide (NO) in many well-known effects of morphine is well defined. NO is involved in the signaling pathway of the N-methyl-D-aspartate (NMDA) receptor, which is proposed to mediate some of morphine's effects. This research studies the effect of morphine and NMDA on lipopolysaccharide (LPS)-stimulated NO production by clonal rat pheochromocytoma (PC12) cells.


METHODS
We used the Griess reaction to measure NO concentrations in cell culture medium.


RESULTS
PC12 cells that were incubated for 24 h with varying concentrations of morphine (0.1, 1, 10, 100, and 1000µM) plus LPS (1 µg/ml) did not significantly alter the concentration of NO in the medium. However, NO production increased when cells were treated for both 48 h with 100 and 1000 µM morphine and for 72 h with 10,100, and 1000 µM of morphine. After 72 h, 1 µM naloxone significantly decreased NO concentration. Naloxone, at doses of 0.1, 1, and 10µM prevented NO production by 1000 µM of morphine. NMDA (0.1, 1, and 10 µM) did not alter NO concentrations after 24, 48 or 72 h. Morphine (1 µM)-induced NO production was inhibited by 10 µM NMDA after 48 h. Inhibition of NO was also noted with1 and 10 µM concentrations of morphine after 72 h.


CONCLUSIONS
Chronic treatment of PC12 cells with morphine significantly increases LPS-stimulated NO production via naloxone-sensitive receptors.The cells seem to release endogenous morphine in medium. NMDA does not affect NO production, which may be due to the lack of functional NMDA receptor expression in PC12 cells.
Department of Neuroscience, School of Advanced Medical Technologies and Department of Pharmacology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
Mohammad Reza
MR
Zarrindast
Mehrak
M
Javadi-Paydar
Ladan
L
Delphi
Nasim
N
Vousooghi
D000701:Analgesics, Opioid / Q000494:pharmacology*; D000818:Animals; D008070:Lipopolysaccharides / Q000494:pharmacology; D009020:Morphine / Q000494:pharmacology*; D016202:N-Methylaspartate / Q000494:pharmacology; D009569:Nitric Oxide / Q000096:biosynthesis*; D016716:PC12 Cells; D051381:Rats
D000701:Analgesics, Opioid; D008070:Lipopolysaccharides; D009569:Nitric Oxide; D016202:N-Methylaspartate; D009020:Morphine