Antisense RNA to inducible nitric oxide synthase reduces cytokine-mediated brain endothelial cell death.
We test whether inhibition of inducible nitric oxide synthase (iNOS) can exert a cytoprotective effect on cerebral endothelial cells upon stimulation by pro-inflammatory cytokines. Mouse brain endothelial cells were stably transfected to express an antisense RNA against iNOS driven by an endothelium-specific von Willebrand factor (vWF) promoter. Upon stimulation with tumor necrosis factor-alpha (TNF-alpha) plus interferon-gamma (IFN-gamma), antisense transfectants showed less iNOS enzymatic activity with less nitric oxide (NO) when compared to the sense control cells. Correspondingly, the antisense cells showed a reduced LDH release and less cytosolic content of oligonucleosomes. These findings establish a cell-specific antisense strategy and confirm the cytotoxic role of iNOS expression in cultured cerebral endothelial cells.
Yang
Ding-I
DI
Institute of Neuroscience, Tzu Chi University, Hualien, Taiwan.
Chen
Shawei
S
Ezekiel
Uthayashanker R
UR
Xu
Jan
J
Wu
Yingji
Y
Hsu
Chung Y
CY
D000818:Animals; D001921:Brain / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D016923:Cell Death / Q000187:drug effects; D002460:Cell Line; D016207:Cytokines / Q000494:pharmacology*; D042783:Endothelial Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D052247:Nitric Oxide Synthase Type II / Q000235:genetics / Q000378:metabolism*; D016372:RNA, Antisense / Q000235:genetics*
D016207:Cytokines; D016372:RNA, Antisense; D052247:Nitric Oxide Synthase Type II