Inhibition of nuclear factor-κB by 6-O-acetyl shanzhiside methyl ester protects brain against injury in a rat model of ischemia and reperfusion.

Recent studies have demonstrated an inflammatory response associated with the pathophysiology of cerebral ischemia. the beneficial effects of anti-inflammatory drugs in cerebral ischemia have been documented. when screening natural compounds for drug candidates in this category,we isolated 6-o-acetyl shanzhiside methyl ester (nd02),an iridoid glucoside compound,from the leaves of lamiophlomis rotata (benth.) kudo. the objectives of this study were to determine the effects of nd02 on a cultured neuronal cell line,sh-sy5y,in vitro,and on experimental ischemic stroke in vivo. for tnf-α-stimulated sh-sy5y cell line experiments in vitro,sh-sy5y cells were pre-incubated with nd02 (20 μm or 40 μm) for 30 min and then incubated with tnf-α (20 ng/ml) for 15 min. for in vivo experiments,rats were subjected to middle cerebral artery occlusion (mcao) for 1 h followed by reperfusion for 23 h. nd02 treatment of sh-sy5y cell lines blocked tnf-α-induced nuclear factor-κb (nf-κb) and iκb-α phosphorylation and increased akt phosphorylation. ly294002 blocked tnf-α-induced phosphorylation of akt and reduced the phosphorylation of both iκb-α and nf-κb. at doses higher than 10 mg/kg,nd02 had a significant neuroprotective effect in rats with cerebral ischemia and reperfusion (i/r). nd02 (25 mg/kg) demonstrated significant neuroprotective activity even after delayed administration 1 h,3 h and 5 h after i/r. nd02,25 mg/kg,attenuated histopathological damage,decreased cerebral evans blue extravasation,inhibited nf-κb activation,and enhanced akt phosphorylation. these data show that nd02 protects brain against i/r injury with a favorable therapeutic time-window by alleviating cerebral i/r injury and attenuating blood-brain barrier (bbb) breakdown,and that these protective effects may be due to blocking of neuronal inflammatory cascades through an akt-dependent nf-κb signaling pathway.