| Objective:With focal cerebral ischemia(MCAO)in rats model of apoplectic cream on angiogenesis in rats after ischemic stroke the mechanism of action,and to explore a Zhongfeng Gaoin the role of angiogenesis in the Wnt signaling pathways for the effects of the process.Methods:Choose healthy adult male SPF SD rats,with improved method to block the left middle cerebral artery of rats,block 2h,the lateral area 2h after ischemia,reperfusion by the left common carotid artery,making rat arterial ischemia reperfusion model(MCAO).After MCAO,the neurological function in rats score,score qualified according to random number table method is divided into model group rats and Zhongfeng Gao group,according to the number of days again divided into 1d,3d,7d and 14d groups.Don't do any deal with blank group,control group in the lower jaw open incision,from rats after careful of left common carotid artery,external carotid artery,internal carotid artery three arterial bifurcation,don't do any processing,small incisions.Blank group and the control measures are not into any intervention until the end of the Zhongfeng Gao group according to the number of days every day Zhongfeng Gao filling and clothing.Not live to the expected number of days in the process of death group added at any time,guarantee the quantity in each group.Nerve function score after MCAO model,and according to the corresponding time points out brain tissue to molecular biological detection.Observation indexes:(1)the Zea-longa nerve function score 5 points score standards;HE staining to observe rat brain ischemia side structure change of the blood vessel cells;Triphenyl four azole nitrogen chloride(TTC)observation rat infarction volume.(2)using immunohistochemical staining observation group rats hippocampus the expression of CD34~+number.(3)by RT-PCR method and Western Blot method of Wnt signaling pathways in?-catenin and Wnt1,Wnt3a,Wnt7b expression were analyzed.Results:1 The Zea-longa 5points 4 score level such as longa nerve function score,observation found that after the building of nerve function score of rats with cerebral ischemia reperfusion slowly with the extension of recovery.Compared with model group,control group of nerve function score were greater than the control group,the difference statistically significant(P<0.01);Compared with Zhongfeng Gao group,model group and Z7,Z14 model group compared with the same time points,statistically significant difference(P<0.05).2 After HE staining were observed under optical microscope(40×10)times,blank group and control group no change in cell structure,shape,quantity;Model 1d ischemia area visible cells in ischemic change,characterized by disorganized,cerebral white matter become loose,but no change,nucleus to the 3d model of ischemia PVS,compared with model group 1d,dyed becomes shallow,pyknotic nucleus;Model 7d to14d,gradually reduce the ischemia performance,14d to the model of brain white matter structure basic returns to normal.Zhongfeng Gao group 1d compared with model 1d group,no significant change;3d stroke compared with model 3d group,from the cell lineage and cerebral white matter structure change has improved significantly.3 The TTC staining to observe the rat brain infarction area after ischemic stroke,and compared with the same time point Zhongfeng Gao found that Zhongfeng Gao group of infarction area compared with the model group were decreased.4 Immunohistochemical method to detect the expression of CD34~+.between the groups with statistical significance(P<0.01),Zhongfeng Gao in the 7d group and model group,expression of CD34~+peak,differences between groups with statistical significance(P<0.01),1d,3d,14d expression of Zhongfeng Gao group compared with model group had a tendency to rise,there was no statistically significant difference between groups(P>0.05).5 In the normal group and the sham operation group,there was a low level expression of?-catenin mRNA,Wnt1 mRNA,Wnt3a mRNA,Wnt7b mRNA,and?-catenin protein,Wnt1 protein,Wnt3a protein and Wnt7b protein.The expression of?-catenin mRNA,Wnt1 mRNA,Wnt3a mRNA,Wnt7b mRNA,?-catenin protein,Wnt1 protein,Wnt3a protein and Wnt7b protein were all increased in rats after ischemic brain injury,with statistical significance(P<0.01).Compared with the model1d and 3d,the rats were compared with the model 1d and 3d.?-catenin,Wnt1,Wnt3a,Wnt7b in mRNA and protein expression of no change,there was no statistically significant difference between the groups(P>0.05),at Zhongfeng Gao group 7d?-catenin,Wnt1,Wnt3a,Wnt7b mRNA and protein expression was compared with model group,the difference was statistically significant(P<0.05),Zhongfeng Gao group 14d?-catenin,Wnt1,Wnt3a,Wnt7b in the mRNA and protein expression in were significantly increased compared with the model group,and the difference was statistically significant(P<0.01).Conclusion:1 Zhongfeng Gao can significantly improve the neurological deficit symptoms in rats with focal cerebral ischemia,and effectively promote the recovery of neurological function after reperfusion.2 Zhongfeng Gao can increase the expression of CD34~+and promote angiog enesis.3 Zhongfeng Gao the mechanism of its action in the process of promoting angiogenesis may by?-catenin and Wnt pathways Wnt1,Wnt3a,Wnt7b related protein expression and work. |
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