| Objective:In this study,micro RNA gene chip technology was applied to screen differentially expressed mi RNAs in the middle cerebral artery occlusion model(MCAO)of rats after electroacupuncture for the treatment of cerebral ischemia.Target gene prediction was performed on differentially expressed mi RNAs,and differentially expressed mi RNAs associated with angiogenesis were screened.GO and KEGG functional enrichment analysis was performed on the predicted target genes by bioinformatics method.To explore the potential target of electroacupuncture in promoting cerebral ischemia.It was revealed that electroacupuncture may regulate angiogenesis by regulating mi RNAs expression changes after cerebral ischemia.Methods:Male adult SD rats of SPF grade were randomly divided into sham-operated group,model group and electroacupuncture group.Rat model of middle cerebral artery occlusion(MCAO)was reproduced with reference to Longa modified method.The electroacupuncture group was selected from the Shuigou point for 30 minutes,and the treatment was continued for 3 days.The cerebral cortex was taken from the ischemic penumbra area at 72 hours after operation.The sham operation group and the model group took the corresponding tissue.(1)Bederson’s five-point method was used to record the degree of neurological deficit in each group of rats,and to observe the effect of electroacupuncture at Shuigou on nerve function in MCAO rats.(2)High-throughput microarray technology was used to analyze the differential expression of mi RNAs in the ischemic penumbra of brain tissue of each group,and differential mi RNA expression profiles were established to screen out differentially expressed mi RNAs.(3)The angiogenesis related mi RNAs were screened,and the correlation between differential mi RNAs and predicted angiogenesis-related target genes was analyzed.Cytoscape software was used to construct mi RNA-m RNA and m RNA-mi RNA-lnc RNA co-expression networks.Results:(1)According to the results of Bederson’s neurological deficit scores,there were no neurological defects in the sham-operated group.Compared with the sham-operated group,the model group had significant neurological deficits.The difference was statistically significant(p<0.01).Compared with the model group,the nerve defect in the electroacupuncture group was alleviated,and the difference was statistically significant(p<0.05).(2)The screening results of differentially expressed mi RNAs related to angiogenesis showed that there were 49 differentially expressed mi RNAs in the model group,of which 37 were up-regulated and 12 were down-regulated.Compared with the model group,there were 21 differentially expressed mi RNAs in the electroacupuncture group,of which 14 were up-regulated and 7 were down-regulated.There were 11angiogenesis-related mi RNAs in the model group vs.sham operation group and the electroacupuncture group vs model group.Among them,the up-regulated genes in the model group were up-regulated in the electroacupuncture group by 8 and down-regulated in 3 groups.(3)After the GO and KEGG functional enrichment analysis of the target genes related to angiogenesis,it was found that the functional annotations of the cerebral ischemia model and the electroacupuncture treatment were mainly as follows: vascular endothelial growth factor receptor binding,ascular endothelial growth factor receptor 2binding,vascular endothelial growth factor binding,regulation of blood vessel endothelial cell migration,Positive regulation of angiogenesis,positive regulation of blood vessel endothelial cell migration.Significant pathways include m TOR signaling pathway,VEGF signaling pathway,T cell receptor signaling pathway,Neurotrophin signaling pathway,Axon guidance,B cell receptor signaling pathway,Erb B signaling pathway.(4)Bioinformatics analysis technology was used to construct mi RNA-m RNA and m RNA-mi RNA-lnc RNA co-expression networks to predict the possible targets of electroacupuncture to regulate angiogenesis after cerebral ischemia.Conclusion:1.The expression profile of mi RNA in the ischemic penumbra region of cerebral tissue of ischemic rats was changed,and the difference was significant,suggesting that mi RNAs may be involved in the gene regulation of the occurrence and development of ischemic stroke.2.Electroacupuncture significantly regulated 21 mi RNAs related to angiogenesis,among which 14 were up-regulated and 7 were down-regulated,suggesting that electroacupuncture may affect the pathophysiology of angiogenesis after cerebral ischemia by regulating mi RNAs. |
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