| ObjectiveBased on the traditional Chinese medicine theory related to "qi and blood--brain pulp" and based on the gene chip detection results of SCF/ c-kit signal pathway in the early stage of the research group,the significantly different results were further explored.The changes of local cerebral blood flow,GRB7 and VEGFR2 on the ischemic side of the model rats were continuously observed at three time points of 3d,7d and 14 d after the intervention treatment with the method of invigorating qi and activating blood circulation and its representative prescription naoluoxintong,to explore the effect of Naoluoxintong on angiogenesis after focal cerebral ischemia and its mechanism.MethodsThe experiment is divided into two parts: experiment 1 and Experiment 2.A total of 96 healthy male SD rats weighing 300±20g were selected and divided into sham operation group(Group J),model group(group M),Tongxinluo group(group T)and Naoluoxintong group(group N)according to the random value table method.There were 24 rats in each group,and 3 subgroups were set up on 3d,7d and 14 d,with 8 rats in each group.A rat model of right middle cerebral artery ischemia-reperfusion(MCAO/R)was established by using the modified thrombus method.After 2h successful completion of modeling,fish lines were drawn for cerebral ischemia-reperfusion,and 2 or 3 points were selected according to Zea Longa evaluation criteria.The doses of naoluoxintong 5.04g/kg and tongxinluo 0.50g/kg were given by gavage twice a day.After the 3d,7d and 14 d,4% paraformaldehyde was perfused in the heart and fixed,and the brain was extracted by craniotomy.Experiment1: IHC Envision two-step method to determine the positive expression areas of GRB7 and VEGFR2 in ischemic frontal parietal cortex of rats in each group.Experiment 2:Laser Doppler ultrasonography was used to detect the changes of r CBF in ischemic cortical infarcts in MCAO/R rats 10 min after successful modeling and after treatment.The results of the experiment,four similar areas were selected from the 400 x field of view of the microscope were analyzed and processed by nanjing jieda DP801 morphological microscopic image analysis system,and IBM SPSS Statistics 24.0software was used to carry out multi-factor anova for repeated measurement of data.The measurement data was represented by Mean±S,and multivariate anova was used for comparison between each group at each time point.was mainly expressed in the nucleus and cytoplasm of nerve cells and endothelial cells in ischemic frontal parietal cortex ResultsExperiment 1.1)Results showed that GRB7 positive expression was brown-yellow in the frontal parietal cortex of the ischemic part,It is w mainly expressed in the nucleus and cytoplasm of nerve cells and endothelial cells in ischemic frontal parietal cortex.There was a small amount of GRB7 expression in group J.Compared with group J,the positive GRB7 expression in group M increased,and the positive area of GRB7 expression was significantly different(P<0.05).Compared with group M,GRB7 expression increased in group N,and the positive expression area was significantly different(P<0.01).Compared with group T,there was no statistically significant difference in GRB7 expression area in group N(P>0.05).1.2)IHC results showed that VEGFR2 protein was brown-yellow in the frontal and parietal cortex of the ischemic part,mainly expressed in the membrane of vascular endothelial cells and the cytoplasm and processes of neurons.There was a small amount of positive expression of VEGFR2 protein in the frontal parietal cortex of the cerebral ischemia side of group J at three different time points,and the positive expression area of VEGFR2 increased in group M compared with group J(P<0.05).Compared with group M,the positive expression area of VEGFR2 protein in group N increased significantly at the same time points,and the differences were statistically significant(P<0.01).There was no significant difference in VEGFR2 expression area between group N and group T(P>0.05).Experiment 2.1 laser doppler detection of focal cerebral ischemia and reperfusion in rats 10 min after successful modeling,compared with group J,the corresponding cerebral blood flow in group M,group T and group N decreased significantly at 3d,7d and 14d(all P< 0.01).There was no statistical difference among the other groups(all P< 0.05).After MCAO/R intervention,cerebral blood flow in group J increased slightly on 3d,7d and14 d,and significantly increased in group M,group T and group N.After treatment,the M group was still significantly lower than the J group,there was no significant difference between group J,group T and group N(P>0.05).Compared with group M,the corresponding three time points in group T and group N were significantly increased(P<0.01).Compared with group T,group N had no statistical significance(P>0.05).ConclusionThis experiment in the early stage of the research method of yiqi huoxue tongluo SCF/c-kit pathway research on the effects of gene expression,on the basis of the positive results of genetic testing further experimental research,explore the SCF/c-kit signaling mechanism,method of yiqi huoxue tongluo and its representative party nao luo xin tong can be expressed by raising GRB7,VEGFR2 protein,adjust the SCF/c-kit signaling pathways,increase the number of new blood vessels after cerebral ischemia,improve cerebral blood flow,reduce brain structure and function injury. |
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