Effect Of Scutellarin And Paeoniflorin Compound On Cerebral Ischemic Injury Rats And Sonic Hedgehog Signaling Pathway

Objective: To investigate the therapeutic effect of Scutellarin and Paeoniflorin compound on permanent cerebral ischemia injury and its possible mechanisms.Methods: Forty-eight adult male Sprague-Dawley rats were divided into four groups randomly: the sham operation group,the cerebral ischemia group,the compound group and the inhibitor group.(n=12 in each group).Permanent middle cerebral artery occlusion(p MCAO)model was performed according to Zea longa's report.Fifteen minutes before ischemia,the inhibitor group was given intraperitoneal injection of Cyclopamine(6mg/kg),the specific inhibitor of sonic hedgehog signaling pathway,while other groups were given equal doses of saline.At 0 hour and 6 hours after ischemia,the compound group and the inhibitor group were given intraperitoneal injection of Scutellarin(20mg/kg)and Paeoniflorin(30mg/kg),while other groups were given equal doses of saline by the same way.Neurological deficit scores were performed at the 24 th hour after ischemia,and then the rats were decapitated.The cerebral infarct volume was detected by 2,3,5-triphenyltetrazolium chloride(TTC)staining.The cell apoptosis was detected by terminal deoxynucleotidyl transfer-mediated d UTP nick end labeling(TUNEL)analysis.RT-PCR and western blotting were used respectively to detect the expression levels of Shh?Patched-1?Gli-1 m RNAs and proteins in the ischemic cortex.Results:(1)The neurological deficit score showed that no neurological deficit was observed in the sham operation group(score 0).The neurological deficit score in the compound group was significantly lower than that in the cerebral ischemia group(P<0.05).Compared with the compound group,the neurological deficit score in the inhibitor group was higher significantly(P<0.05).(2)TTC staining showed that no infarct was observed in the sham operation group.The infarct volume in the compound group was significantly lower than that in the cerebral ischemia group(P<0.05).Compared with the compound group,The infarct volume in the inhibitor group was higher significantly(P<0.05).(3)The TUNEL staining showed that the apoptosis rate in the cerebral ischemiagroup increased significantly compared with that in the sham operation group(P<0.05).The apoptosis rate decreased significantly in compound group compared with that in the cerebral ischemia group(P<0.05).The apoptosis rate was increased significantly in inhibitor group than that in the compound group(P<0.05).(4)RT-PCR result suggested that the m RNA levels of Shh,Patched-1 and Gli-1 in the cerebral ischemia group were significant higher than that in the sham operation group(P<0.05)and significant lower than that in the compound group(P<0.05).The m RNA level of GLI-1 in the inhibitor group was lower significantly than in the compound group(P<0.05).(5)Western blotting result showed that the protein expression levels of Shh,Patched-1 and Gli-1 in the cerebral ischemia group were higher significantly than that in the sham operation group(P<0.05)and lower significantly than that in the compound group(P<0.05).The expression level of GLI-1 protein in the inhibitor group was lower significantly than that in the compound group(P<0.05).Conclusion: The compound of Scutellarin and Paeoniflorin has certain therapeutic effect on permanent cerebral ischemia injury.Its mechanism is associated with the activation of SHH signaling pathway.