| Objective: In this experiment,from the perspective of bioinformatics,based on the geo database to screen the serum biomarkers of rats with cerebral infarction,and through animal experiments to explore the effect of Huoxue rongluo formula on the serum BDNF and b FGF of rats with MCAO reperfusion injury.Method:In the first part,based on the geo database,the serum biomarkers of rats with cerebral infarction were screened:(1)using the geo database to download the blood chip of gse119121 rats,and using the R language analysis to get the differential genes.(2)Using the string functional protein association database,we constructed and focused the PPI network of cerebral infarction differential genes,and used the degree,closeness,betweenness algorithm topology analysis.(3)Go function enrichment and KEGG pathway enrichment of cerebral infarction differential genes were carried out by using webgestalt and David databases.In the second part,the effects of Huoxue rongluo Recipe on serum BDNF and b FGF in MCAO reperfusion injury rats were studied.120 male SD rats were randomly divided into four groups: sham operation group(sham group,n = 30),model group(MCAO group,n = 30),Huoxue rongluo recipe group(MCAO + Huoxue rongluo recipe group,n = 30),butylphthalide group(MCAO + butylphthalide group,n = 30).Each group was divided into three subgroups: 3D,7d and 14 d 10 rats in each subgroup.The model of MCAO reperfusion injury was established and the neurological deficit was evaluated.The rats were given the first gavage(1ml / 100g)6 hours after anesthesia.On the 3rd,7th and 14 th day after reperfusion,the neurological deficit was scored;on the 7th day after reperfusion,he staining was used to observe the pathological changes of brain tissue;on the 3rd,7th and 14 th day after reperfusion,the expression of b FGF and BDNF in serum was detected by ELISA kit.Result:In the first part,based on the geo database,the serum biomarkers of cerebral infarction rats were screened:(1)the blood chip of gse119121 rats was retrieved,and 271 differential genes were screened by R language,among which 102 genes were down-regulated and 169 genes were up-regulated(including BDNF and b FGF).(2)The PPI of differential gene protein was constructed and focused by using string database and Cytoscape software,and the topological analysis of degree,betweenness and closeness algorithm was carried out.(3)Using webgestalt database,271 differentially expressed genes were analyzed for go function enrichment.Through the analysis of KEGG signal pathway enrichment of 271 differential genes by David database,it was found that the main enrichment was in hematopoietic cell lineage,the interaction between cytokines and cytokine receptor,primary immune deficiency,MAPK signal pathway,etc.In the second part,the effect of huoxuerongluo formula on the serum BDNF and b FGF of rats with MCAO reperfusion injury was discussed:(1)the score of neural function defect: there was no neural function defect in the sham operation group;compared with the model group,the score of Shenjing function defect in the huoxuerongluo formula group and the butylphthalide group was significantly reduced 3D,7d and 14 d after reperfusion(P<0.01).(2)He staining results: under the light microscope(× 400),in the sham operation group: the morphology of neurons in the cerebral cortex of the ischemic side of the rats was normal,the arrangement was orderly and dense,the cells were round or oval,and there was no obvious swelling;in the model group: a large number of neurons in the cerebral cortex of the ischemic side of the rats were degenerated,atrophied,the arrangement of cells was disordered,the vacuolation of cells was obvious,and the boundary between nucleus and cytoplasm was obvious Compared with the model group,Huoxue rongluo formula group and butylphthalide group can improve the morphology of neurons in cerebral cortex of ischemic side in varying degrees.There are a few vacuole like changes in nerve cells,the arrangement is still neat,and the peripheral interstitial damage is light.(3)Results: compared with the sham operation group,the content of BDNF and b FGF in the model group,huoxuerongluo group and butylphthalide group increased significantly(P < 0.01)on the 3rd,7th and 14 th day after reperfusion,and the content of BDNF and b FGF in the model group and butylphthalide group decreased significantly(P < 0.01)on the 3rd,7th and 14 th day after reperfusion.Conclusion:In the first part,using bioinformatics to analyze the original gene chip data of geo database,we can effectively screen the serum biomarkers of rats with cerebral infarction,reveal the possible pathogenesis of cerebral infarction,and suggest that it is related to multiple targets and multiple signal pathways,which provides a new idea for future clinical research.In the second part,we found that the expression of BDNF and b FGF in the serum of MCAO rats was up-regulated,which was consistent with the data obtained by bioinformatics analysis.Huoxue rongluo recipe could promote the recovery of nerve function,alleviate the pathological damage of brain tissue,and down regulate the expression of BDNF and b FGF in the serum of rats with cerebral ischemia-reperfusion injury.We speculated that its mechanism might be related to the improvement of BBB Reply. |
PDF Full Text Request