Effect Of Electroacupuncture Pretreatment At Jiaji Point In Rats With Myocardial Ischemia Reperfusion Injury Based On RNA-seq

OBJECTIVE: In this study,the protective effects of preconditioning with electroacupuncture at the Jiaji acupoint T4 and T5 on myocardial ischemia-reperfusion injury in rats were studied,and the gene expression profile of electroacupuncture at Jiaji points was performed by RNA-seq(RNA sequencing)technique.Analysis,screening differentially expressed m RNA and signaling pathway to explore the protective mechanism of electroacupuncture at Jiaji points for reperfusion injury.METHODS: Adult Wistar male rats were randomly divided into control group,model group,Jiaji group and Neiguan group,with 12 rats in each group.The control group and the model group were bundled for 30 min each time,once a day for 7 days.The electroacupuncture pretreatment group was acupunctured for 30 min each time,once a day,and the electroacupuncture was continued for 7 days.The electroacupuncture uses a sparse wave(2/100 Hz)with a 1 m A intensity.On the second day of preconditioning,a model of myocardial ischemia-reperfusion injury was established by ligation of the left anterior descending coronary artery for 30 min and reperfusion for 24 h.The control group was only threaded without ligation.During the experiment,the electrocardiogram of the II lead was monitored and the changes of TII electrocardiogram were analyzed to evaluate the occurrence of arrhythmia.After the end of the animal experiment,the serum of each group was taken to detect CK-MB and c Tn I.The heart of the rat was observed by HE staining.Morphological changes,ultrastructure of cardiomyocytes were observed under transmission electron microscope;myocardial infarct size was determined by TTC-Evans blue staining of rat hearts;4 hearts in each group were taken from left ventricular ischemic area to extract total RNA,and c DNA was constructed.After the library,16 samples were sequenced by RNA-seq technique,and the differentially expressed m RNA was screened.The functional clusters of differentially expressed m RNA were analyzed by GO analysis.The signal pathway of differentially expressed m RNA was analyzed by KEGG enrichment.The biological effects and signal transduction pathways involved in differentially expressed m RNA were preliminarily explored.At the same time,the weighted gene co-expression network(WGCNA)was used to analyze the differentially expressed m RNA of the Jiaji group.RESULTS: 1.The myocardial ischemia-reperfusion injury model was successfully established by ligation of TII electrocardiogram,serum CKMB,c Tn I value and myocardial pathological morphology for 30 min of left anterior descending coronary artery in rats.2.Compared with the model group,electroacupuncture at Jiaji and Neiguan acupoints can significantly reduce the release of serum myocardial enzymes CK-MB and c Tn I in rats with myocardial ischemia-reperfusion injury,reduce the TII wave height of ECG,and reduce the occurrence of reperfusion arrhythmia.Reduce myocardial infarct size and reduce myocardial pathological damage.3.The transcriptome of left ventricle myocardium was successfully constructed by RNA-seq technique.The differential gene cluster analysis showed that the gene expression patterns of myocardial tissue in the control group,model group,Jiaji group and Neiguan group were significantly changed..4.Differentially expressed m RNA analysis showed that there were 7419 differentially expressed m RNAs in the model group compared with the control group,of which 3694 were up-regulated and 3,725 were down-regulated,indicating that the m RNA of rats with myocardial ischemia-reperfusion injury changed significantly;Compared with the model group,there were 1243 differentially expressed m RNAs,of which 678 were up-regulated and 565 were down-regulated.There were 304 differentially expressed m RNAs between the Neiguan and model groups,of which 165 were up-regulated and 139 were down-regulated.The results showed that there were significant changes in m RNA expression in myocardial ischemia-reperfusion injury after electroacupuncture at Jiaji and Neiguan.The differentially expressed m RNA in the Jiaji group was more than that in the Neiguan group.5.GO analysis showed that there were more GO functional items in the model group,the Jiaji group and the Neiguan group.The model group was mainly enriched in the small molecule metabolic process,the regulation of multicellular biological processes,and the biological processes.Positive regulation,etc.;the Jiaji group is mainly concentrated in the defense response,immune system process,innate immune response,immune response,regulation of the immune system process.The Neiguan group is mainly enriched in defense responses,immune system processes,and responses to biological stimuli.6.KEGG enrichment analysis revealed that the model group differentially expressed m RNA enrichment in oxidative phosphorylation,valine,leucine and isoleucine degradation,retrograde endogenous cannabinoid signaling,chemokine signaling pathway,etc.Strip access.The differentially expressed m RNA in the Jiaji group is enriched in multiple pathways such as extracellular matrix receptor interaction,focal adhesion,and chemokine signaling pathway.There is no enrichment in the Neiguan group.CONCLUSION:1.Pretreatment with electroacupuncture at Jiaji has protective effects on myocardial ischemia-reperfusion injury in rats,which can significantly reduce the release of serum myocardial enzymes CK-MB and c Tn I,reduce the high T wave of ECG,reduce the occurrence of arrhythmia,and reduce myocardial damage..2.Multiple signal transduction,immune regulation,substance and energy metabolism and other related genes and pathways involved in myocardial injury during myocardial ischemia-reperfusion injury.Electroacupuncture at Jiaji points is involved in the regulation of multiple genes in myocardial injury.The expression of the pathway,and the expression pattern has changed significantly.