| Objective:To study on effect of Danlong xingnao recipe on proliferation of endogenous neural stem cells(NSCs) and expression of Wnt-1, Wnt-3a, β-catenin and GSK-3β in hippocampus of cerebral ischemia-reperfusion (I/R) injury modle rats.Methods:60male SD rats were randomly divided into sham group, cerebral ischemia reperfusion injury (model) group (I/R), edaravone (western medicine) group, Danlong Xingnao recipe small dose (Dan Xiao) group, Danlong Xingnao recipe high-dose (Dan Da) group,12rats in each group.In addition to the sham group, The focal Middle cerebral artery occlusion (MCAO) injury models were prepared by suture method, and recoved of2h reperfusion.Sham group and modle group were given capacity distiled water, western medicine group were given3.2mg kg-1dose intraperitoneal to medicine, Dan Xiao group and Dan Da group were given respectively3.7g kg-1,7.4g kg-1doses intragastric to medicine,1time every day, for7consecutive days, take rats brain issue ischemia side in the last2h after treatment.(1) Observe the neurological score changes of rat;Nissl stained method is used to observed the pathomorphological changes in hippocampus CA1area, and recorded the positive cells number with nissl body;(2)BrdU incorporation method is used to record the BrdU positive cells number of hippocampus dentate gyrus;(3)Took the RT-qPCR method to detect hippocampal Wnt-1, Wnt-3a, β-catenin, GSK-3β mRNA expression; (4)Using Western Blot method to detect the hippocampus Wnt-1, Wnt-3a, β-catenin, cytoplasm β-catenin, GSK-3β protein expression.Results:(1) Compared with sham group, neurological score enhanced obviously in other groups, the difference was statistically significant (P<0.01); Compared with model group, neurological score reduced obviously in western medicine group, Dan Xiao group, Dan Da group, the difference was statistically significant (P<0.05, P<0.01);(2)Compared with sham group, the positive cells number with nissl body, Brdu positive cells, Wnt-1, Wnt-3a, β-catenin protein and mRNA expression enhanced obviously, GSK-3β protein and mRNA expression reduced obviously in other groups, the difference was statistically significant (P<0.05, P<0.01); Compared with model group, the positive cells number with nissl body,Brdu positive cells, Wnt-1, Wnt-3a, β-catenin, cytoplasm β-catenin protein and mRNA expression improved obviously, GSK-3β protein and mRNA expression reduced obviously in western medicine group, Dan Xiao group, Dan Da group, the difference was statistically significant (P<0.05, P<0.01).Conclusion:(1)Danlong Xingnao recipe can promote the neurological function recovery by reducing the neurons loss, and relieved cerebra ischemia injury;(2)Danlong Xingnao recipe may activate the Wnt/β-catenin pathway by increasing Wnt-1, Wnt-3a,β-catenin expression, inhibiting GSK-3β expression, promoting the gather of β-catenin in cytoplasm, and promote proliferation of endogenous neural stem cells in hippocampus of cerebral ischemia reperfusion injury modle rats. |
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