Synergistic Protective Effect Of Hydroxysafflower Yellow A And Paeonifolrin On NF-κB Pathway In Cerebral Ischemia Reperfusion Injury Rats

Purpose1.To observe the effect of pharmacological conditioning on neurological function and cerebral infarction area in rats with cerebral ischemia reperfusion injury,and to explore the synergistic protective effect of hydroxysafflor yellow A and paeoniflorin on the brain tissue of rats.2.To observe the effect of hydroxysafflor yellow A and paeoniflorin on the expression of IL-1β,TNF-α rats with cerebral ischemia reperfusion injury,and to explore the effect of hydroxysafflor yellow A and paeoniflorin on the expression of inflammatory factors.3.To observe the effect of hydroxysafflor yellow A and paeoniflorin on the expression of p65 in the NF-κB signaling pathway,and to investigate the molecular mechanism of hydroxysafflor yellow A and paeoniflorin for anti-inflammation.Method120 SPF level male SD rats,weighed(200 ± 20)g,were randomly divided into six groups:sham operation group,model group,positive(gingko lactone)group(5mg/kg),hydroxysafflower yellow A group(5mg/kg),paeoniflorin group(5mg/kg)and hydroxysafflower yellow A combined with paeoniflorin group(5mg/kg +5mg/kg),20 rats in each group.After 3 days of adaptive feeding,the rat model of acute focal cerebral ischemia was established by using the right middle cerebral artery occlusion method(MCAO)and the neurological deficit score was performed and drugs were administrated by tail intravenous injection.Neurological deficit was revalued after 7 days treatment.The area ratio of cerebral infarction in rats was measured by triphenyltetrazolium chloride(TTC)staining.The level of IL-1β and TNF-a in cerebral homogenate were examined by Enzyme linked immunoassay(ELISA).The expression of NF-κB were examined by using immunohistochemistry.Result1.Compared with the sham-operated group,the scores of neurological deficits in the model group were increased(P<0.05).Compared with the model group,the neurological scores of drug-treated groups were significantly decreased(P<0.05).There was no statistically significant difference between each drug group.(P>0.05).2.Compared with the sham-operated group,the area ratio of cerebral infarction in rats with cerebral ischemia reperfusion was significantly higher than that in the model group(P<0.05).Compared with the model group,the cerebral infarction area ratios of drug-treated groups were significantly decreased(P<0.05),while the hydroxysafflower yellow A combined with paeoniflorin group was significantly better than other groups.3.Compared with the sham-operated group,the content of IL-1β and TNF-αmodel group was significantly increased(P<0.05).Compared with the model group,the level of IL-1β and TNF-α of rat brain tissue in drug-treated groups were significantly decresed(P<0.05).Among them,the hydroxysafflower yellow A combined with paeoniflorin group seemed to have a better effect inhibitory effect on the expression of IL-1β and TNF-α than in other groups.4.Compared with the sham-operated group,the expression of NF-κB in the model group were increased(P<0.05).Compared with the model group,the expression of NF-κB were all attenuated in four drug-treated groups(P<0.05).Meanwhile,compared with the combination of hydroxysafflor yellow A and paeoniflorin group,the expression of NF-κB were increased in other three groups(P<0.05).Conclusion1.The treatment of hydroxysafflor yellow A and paeoniflorin significantly reduced the neurological function score and the area of cerebral infarction after cerebral ischemia-reperfusion injury,indicating that hydroxysafflor yellow A and paeoniflorin have a therapeutic cerebral ischemia-reperfusion injury,and the combination effect of them was more significant.2.The treatment of hydroxysafflor yellow A and paeoniflorin can significantly reduced the content of IL-1β and TNF-αin cerebral homogenate,indicating hydroxysafflor yellow A and paeoniflorin treatment could inhibit excessive activation of inflammatory factors.And the effect of the combination of hydroxysafflor yellow A and paeoniflorin is the best.3.The treatment of hydroxysafflor yellow A and paeoniflorin can significantly reduced the overexpression of NF-KB/p65 in hippocampal CA1 area of rats after cerebral ischemia-reperfusion injury,and the combination effect of them was more significant,suggesting that the combination of HSYA and paeoniflorin may alleviate the encephalitis reaction by intervening NF-kappa B pathway.