The Regulation Of Ilexonin A On Activation Of Canonical Wnt Signaling And Neural Regeneration After Cerebral Ischemia-Reperfusion In Rats

OBJECTIVE：To observe the regulation of Ilexonin A（IA） on canonical Wntsignaling and neural regeneration after cerebral ischemia reperfusion in rats and itspossible mechanism.METHODS：One hundred and forty-four Sprauge-Dawley male rats wererandomly divided into normal group, sham group(The procedure with the modelgroup, but the middle cerebral artery is not blocked), model group, IA group, eachgroup divided into1d、3d、7d、14d four subgroups, nine rats of each group. The modelof focal ischemia reperfusion was made by the middle cerebral artery occlusionthrough line plug, Neuronogic impairment were assessed through the method ofLonga scoring; TTC staining was used to view the infarct of ischemia reperfusion;Immunofluorescent double staining method was applied to detect Brdu/nestin、Brdu/NeuN double positive cells in the perienchyma of ischemic tissue;Immunofluorescence staining、western blotting and RT-PCR were used to observe theexpression of β-catenin、GSK3β、Axin、LEF-1of canonical Wnt signalingpathways related factors.RESULTS：（1）The symptom of neurologic impairment were obvious at1d afterreperfusion, The symptom of neurologic impairment were gradually repaired with theincrease of time. The neurologic impairment score of the treatment group was lessthan model group, The treatment group’s neurologic impairment score was significantdifference with model group at3d、7d（P<0.05）.（2）The level of positive cells inthe perienchyma of ischemic tissue: The model and treatment group’s Brdu/nestin、Brdu/NeuN double positive cells obvious increased and reached its peak at3d, itbegan to decline at7d and still expression at14d, A significant difference in treatmentgroup compared with model group at3d、7d（P<0.05）.（3）The level of β-cateninpositive cells in the perienchyma of ischemic tissue: The model group’s β-cateninpositive cells increased at1d and reached its peak at3d, it began to decline at7d andstill expression at14d, The β-catenin positive cells of the treatment group was morethan model group, A significant difference at3d、7d in treatment group compared with model group（P<0.05）（.4）Western blotting：①The expression of β-catenin protein:The model group’s β-catenin protein increased at1d and reached its peak at3d, itbegan to decline at7d and still expression at14d, The β-catenin protein of thetreatment group was more than model group, A significant difference at1d、3d、7d intreatment group compared with model group（P<0.05）. The protein expression of β-catenin has the similar result with the Immunofluorescence.②The expression ofGSK3β protein: The model group’s GSK3β protein declined with the prolongationof time, The GSK3β protein of the treatment group was less than model group, Asignificant difference at1d、3d、7d、14d in treatment group compared with modelgroup（P<0.05）.（5）RT-PCR detection method:①The expression of Axin mRNA:The model group’s Axin mRNA declined with the prolongation of time, The AxinmRNA of the treatment group was less than model group, A significant difference at3d、7d in treatment group compared with model group（P<0.05）.②The expressionof LEF1mRNA: The model group’s LEF1mRNA increased at1d and reached itspeak at3d, it began to decline at7d and still expression at14d, The LEF1mRNA ofthe treatment group was more than model group, A significant difference at3d、7d、14d in treatment group compared with model group（P<0.05）.CONCLUSION：IA can significantly repair the neurological deficits aftercerebral ischemia reperfusion in rats. It may be pass thought enhance the level of β-catenin protein and promote β-catenin protein into the nuclear, decline theexpression of inhibitor factor GSK3β、Axin, activate the target gene LEF1topromote Wnt signaling to promote neural regeneration and to protect the damagedbrain tissue.