Effect Of Tilianin On The Angiogenesis After Stroke In Brain Tissue Of Rats

Objective: The aim of our study is to establish Middle Cerebral Artery Occlusion(MCAO)rat model to investigate the effect of tilianin on blood circulation ischemic area after stroke in rats and its possible mechanism.Meanwhile,to further study provide experimental evidence.Methods:(1)Establish MCAO rat model.(2)Neuro-functional scoring and grouping,the groups are randomlydivided into model group(normal saline gavage),tilianin low(4mg/kg),middle(8 mg/kg)and high(16 mg/kg)groups and sham-operated group(normal saline gavage,only ligation of the external carotid artery).(3)Triphenyl Tetrazolium Chloride(TTC)staining is used to detect the acreage of cerebral infraction in rats.(4)The number of blood vessels that maintain perfusion in the ischemic cortex was detected by the latex perfusion method.(5)The immunofluorescence(IF)method is used to detect microvascular density around the infarct and ki67 labeled neovascularization number.(6)The expression level of VEGF,Ang-1,p-Src/Src and p-Erk/Erk in brain tissue are detected by Western Blot(WB).Results:(1)Compared with the model group,the neurological scores of the in medium dose and high dose groups of tilianin were significantly decreased(P<0.05,P<0.01).(2)TTC staining results showed that the infarct acreage of model group was 27.96% of the whole brain.Compared with model group,tilianin middle-dosage of infarction acreage group was 11.73% and high-dose of infarction acreage group was 6.04%(P<0.05,P<0.01).(3)After 7 days of pre-administration of tilianin,the numbers of blood vessels could maintain blood flow in the tilianin middle and high dose groups,which were higher than those in the model group,and the proportion of the tiny vessels in the preserved blood vessels was higher than that in the model group.(4)IF results showed that the area ratio of blood vessels in the tilianin middle dose and the high dose groups was higher than the model group,and the number of ki67 labeled blood vessels was significantly higher than that of the model group(P<0.01,P<0.01).(5)WB results showed that the protein expression levels of VEGF,Ang-1,p-Src and p-Erk in the tilianin middle dose and the high dose group were significantly increased(P<0.05).Conclusion: Tilianin could effectively promote angiogenesis after stroke.Its mechanism may regulate of p-Src,p-Erk and other proteins downstream of VEGF signaling pathway.