Antisense blockade of inducible nitric oxide synthase in glial cells derived from adult SJL mice.
Increasing evidence suggests a correlation between cytokine-induced nitric oxide synthase (iNOS) and demyelination in Multiple sclerosis (MS). Inhibition of iNOS may therefore be a novel therapeutic approach in MS. To test an antisense oligodeoxynucleotide (ODN) knockdown strategy for inhibiting iNOS, we used lipopolysaccharide (LPS) together with gamma-interferon (IFN-gamma) to induce iNOS in adult mouse mixed glial cell cultures. We administered an iNOS-derived antisense phosphorothiorate oligodeoxynucleotide (S-ODN) to block the induction. The antisense ODN treatment resulted in significant inhibition of LPS and IFN-gamma induced iNOS mRNA and protein expression. It also inhibited nitric oxide (NO) and cyclic GMP (cGMP) production in a dose dependent fashion. Sense and random S-oligo had no effect in any of these studies. These data indicate the efficacy and specificity of the antisense oligodeoxynucleotide approach in inhibiting iNOS in glial cells.
Ding
M
M
Department of Neurology, School of Medicine, University of California at Los Angeles, Reed Neurological Research Center 90095-1769, USA. mding@ucla.edu
Zhang
M
M
Wong
J L
JL
Voskuhl
R R
RR
Ellison
G W
GW
D000818:Animals; D001483:Base Sequence; D006152:Cyclic GMP / Q000037:antagonists & inhibitors; D007371:Interferon-gamma / Q000494:pharmacology; D008070:Lipopolysaccharides; D051379:Mice; D008969:Molecular Sequence Data; D009103:Multiple Sclerosis / Q000201:enzymology* / Q000628:therapy; D009457:Neuroglia / Q000378:metabolism; D009569:Nitric Oxide / Q000037:antagonists & inhibitors; D019001:Nitric Oxide Synthase / Q000378:metabolism*; D016376:Oligonucleotides, Antisense / Q000494:pharmacology* / Q000627:therapeutic use
D008070:Lipopolysaccharides; D016376:Oligonucleotides, Antisense; D009569:Nitric Oxide; D007371:Interferon-gamma; D019001:Nitric Oxide Synthase; D006152:Cyclic GMP