This study is to investigate the effect of hydroxyethylpuerarin on the expression of
tumor necrosis factor-alpha (
TNF-alpha) and activity of
nuclear factor kappa B (
NF-kappaB) after
middle cerebral artery occlusion (MCAO) in
rats.
Rats were subjected to
cerebral ischemia-
reperfusion injury induced by MCAO. Hydroxyethylpuerarin (10, 20, 40 mg x kg(-1), iv) was administered just 30 min before occlusion and immediately after
reperfusion. After a 24 h
reperfusion following 2 h of MCAO, the number of viable
neurons in
hippocampal CA1 region was counted by
hematoxylin and
eosin (HE)
staining.
TNF-alpha protein and its
mRNA expression were examined with
radioimmunoassay (RIA) and reverse transcriptasepolymerase
chain reaction (RT-PCR) respectively.
NF-KB activity was observed by
electrophoretic mobility shift assay (EMSA), and inhibition of
NF-kappaB alpha (
IkappaBalpha)
protein expression was evaluated by
Western blotting analysis.
Animals treated with hydroxyethylpuerarin had a significant increase in neuronal
survival in comparison with vehicle-treated group. Hydroxyethylpuerarin significantly reduced the
protein and
mRNA expression of
TNF-alpha following 2 h of
ischemia with 24 h of
reperfusion.
NF-kappaB DNA binding activity and the degradation of
IkappaBalpha in the
cytoplasm also decreased by hydroxyethylpuerarin
treatment. The protective effects of hydroxyethylpuerarin against
ischemia-reperfusion injury may be mediated by decreasing the expression of
TNF-alpha and the activity of
NF-kappaB in
rats.