The Effects Of Two Traditional Chinese Medicine Treatments On Angiogenesis And SCF/c-kit Signaling Pathway In Rats With Focal Cerebral Ischemia And Reperfusion

Objective This experiment is based on the brain-related and blood-related theory of ischemic stroke,based on previous research work and establish a rat model of focal cerebral ischemia and reperfusion,application of two kinds of experiences in treatment of Wang's internal medicine(Yiqi Huoxue Tong Luo method,Bushen Shengsui method)and its representative of Chinese medicine compound Objective To observe the changes of the angiogenesis after cerebral ischemia and SCF/C-kit signaling regulatory pathway.Used Immunohistochemistry two-step and Western blot detection method to detect the angiogenesis of the ischemic cortex and the expression of rat cerebral cortex SCF?C-kit?CD34?VEGF and Ang-1,To explore the significance of angiogenesis after cerebral ischemia,The Role of SCF/C-kit Signaling Pathway in Cerebral Ischemia Regeneration and the Effects of Two TCM Treatments on Vascular Regeneration and SCF/C-kit Signaling Regulatory Pathways and Their Mechanisms of Action,to provide experimental basis for the clinical rational application of two traditional Chinese medicine prescriptions.Methods Healthy male SD(Sprague-Dawley)rats 72,age 4m,weight 280~320g.The rats were randomly divided into four group: sham operation group,cerebral ischemia model group,Yiqi Huoxue Tong Luo method group,Bushen Shengsui method group.The model of MCAO/R rat was established by reforming longa method.The score of neurological deficit was 2 or 3.The brain tissue of each group of rats was fixed and coronal brain sections were taken.Using immunohistochemistry envision two-step to detect Proteins Changes in positive area and average absorbed optical density of CD34,VEGF,Ang-1 in the parietal cortex of ischemic side of rats in each group.Western Blot method was used to detect the relative gray value changes of SCF and C-kit protein expression in hippocampus and fronto-parietal cortex of four groups of rats.DP801 morphology microscopic image analysis system was used for immunohistochemical image analysis,Western Blot analysis was performed using the AIC.Alpha View system,count and record all data.Using spss18.0 for finishing analysis,measurement data was expressed using Mean±S,The comparison of mean values at multiple time points between multiple groups was compared using repeated measurement variance analysis.If the data covariance matrix satisfies the spherical test,the sphericity test is used.If not,Huynh-feldt correction is used;One-way analysis of variance was used to compare different groups at the same time;If the difference between the populations is statistically significant,use LSD for comparison,The same group at different time points were compared using the univariate repeated measures analysis of variance,If the overall difference between the groups is statistically significant,Bonferroni(B)is used for post-mortem comparisons;assume that the test level is set to 0.05.Use EXCEL2007 software to make statistics charts and tables.Results1)Effects of two traditional Chinese medicine treatments on Expression of CD34,Ang-1 and VEGF Protein in the Cortex of the Parietal Cortex of Rats.Comparison of three protein expression levels in different groups after reperfusion for3 days: After analysis of variance,there was a statistically significant difference between the four groups(P<0.01),after a couple of comparisons,the model group was lower than the sham group,the Yiqi Huoxue Tong Luo method group and the Bushenshengsui method group were higher than the sham group,the Yiqi Huoxue Tong Luo method group and the Bushen Shengsui method group were higher than the model group,the Yiqi Huoxue Tong Luo method group was higher than the Bushenshengshengsui method group,The difference was statistically significant(P<0.01);Comparison of three protein expression levels in different groups after reperfusion for 7 days: After analysis of variance,there was a statistically significant difference between the four groups(P<0.01),after a couple of comparisons,the model group was lower than the sham group,the Yiqi Huoxue Tong Luo method group and the Bushenshengsui method group were higher than the sham group,the Yiqi Huoxue Tong Luo method group and the Bushen Shengsui method group were higher than the model group,the difference was statistically significant(P<0.01),but there was no significant difference between the Yiqi Huoxue Tong Luo method group and the Bushenshengsui method group;Comparison of three protein expression levels in different groups after reperfusion for14 days: After analysis of variance,there was a statistically significant difference between the four groups(P<0.01),after a couple of comparisons,the model group was lower than the sham group,the Yiqi Huoxue Tong Luo method group and the Bushenshengsui method group were higher than the sham group,the Yiqi Huoxue Tong Luo method group and the Bushen Shengsui method group were higher than the model group,the Bushen Shengsui method group was higher than the Yiqi Huoxue Tong Luo method group.The difference was statistically significant(P<0.01);The univariate repeated-measures analysis of variance was used to compare the positive areas of the three proteins in the parietal cortex at different time points within the group.There was no significant difference in sham operation group,model group,Yiqi Huoxue Tong Luo method group,and Bushen Shengsui method group at different time(P>0.05).There was no statistically significant in average absorbed optical density.2)Effects of two traditional Chinese medicine treatments on Expression of SCF and C-kit Protein in Frontal Parietal Cortex of Model Rats.In the parietal cortex,Comparison of expression levels of SCF and C-kit after 3 days of reperfusion in different groups: After analysis of variance,there was a statistically significant difference between the four groups(P<0.01),after a couple of comparisons,the model group was lower than the sham group,the Yiqi Huoxue Tong Luo method group and the Bushenshengsui method group were higher than the sham group,the Yiqi Huoxue Tong Luo method group and the Bushen Shengsui method group were higher than the model group,the Yiqi Huoxue Tong Luo method group was higher than the Bushenshengsheng sui method group,The difference was statistically significant(P<0.01);Comparison of expression levels of SCF and C-kit after 7 days of reperfusion in different groups: After analysis of variance,there was a statistically significant difference between the four groups(P<0.01),after a couple of comparisons,the model group was lower than the sham group,the Yiqi Huoxue Tong Luo method group and the Bushenshengsui method group were higher than the sham group,the Yiqi Huoxue Tong Luo method group and the Bushen Shengsui method group were higher than the model group,the Bushenshengshengsui method group was higher than the Yiqi Huoxue Tong Luo method group,the difference was statistically significant(P<0.01);Comparison of expression levels of SCF and C-kit after 14 days of reperfusion in different groups: After analysis of variance,there was a statistically significant difference between the four groups(P<0.01),after a couple of comparisons,the model group was lower than the sham group,the Yiqi Huoxue Tong Luo method group and the Bushenshengsui method group were higher than the sham group,the Yiqi Huoxue Tong Luo method group and the Bushen Shengsui method group were higher than the model group,the Bushenshengshengsui method group was higher than the Yiqi Huoxue Tong Luo method group,the difference was statistically significant(P<0.01);The variance analysis of SCF and C-kit at different time points within the group was performed using the univariate repeated measures analysis of variance.There was no significant difference between the sham operation group,the model group,the Yiqi Huoxue Tong Luo method group and the Bushen Shengsui method group at different time points(P>0.05).Conclusion Both treatments regulate the SCF/C-kit signaling pathway by promoting the expression of CD34,VEGF,and Ang-1 proteins,Promoting blood vessel regeneration after cerebral ischemia,it is beneficial to the damage and repair of damaged brain tissue after focal cerebral ischemia and reperfusion.