| backgroundAtrial fibrillation(AF)is one of the most common arrhythmia in clinic.Its incidence rate,incidence rate and morbidity are increasing,which brings burden to millions of people around the world.Therefore,both clinical research and basic research have become a hot topic in cardiovascular research.Atrial fibrillation is characterized by the replacement of regular atrial electrical activity by rapid atrial fibrillation,which leads to the disorder of atrial rhythm and the deterioration of mechanical function.Long term maintenance of atrial fibrillation can lead to ventricular rhythm disorder,cardiac dysfunction and atrial thrombosis,and increase the risk of heart failure,stroke,thromboembolism and so on,which is very harmful to the life safety of patients.Epidemiological findings showed that the incidence rate of atrial fibrillation in the general population was about 1-2%,and gradually increased with age,reaching 10% over 80 years old.The main pathological features of AF are electrical remodeling,structural remodeling and neural remodeling,but the exact pathogenesis of AF is still unclear.In recent years,it has been found that atrial fibrillation related RNA plays an important role in the pathophysiological process of atrial fibrillation by participating in the inflammatory response,intervening in the formation of ion channels,regulating the degradation balance of extracellular matrix and so on.Therefore,it is of great clinical significance to study the m RNA and signal pathway related to the pathogenesis of atrial remodeling based on this level,which is expected to provide a new strategy for the prevention and treatment of atrial fibrillation.objectiveA-TP was used to establish the model of experimental atrial fibrillation in goats.The second-generation deep sequencing technology was used to detect the differential expression of messenger ribonucleic acid(m RNAs)transcripts in the right atrium of the experimental group and the control group Analysis)and other bioinformatics methods to explore the m RNAs and cellular signal transduction pathways related to atrial fibrillation remodeling.Method1.Establishment of goat atrial fibrillation modelFourteen healthy adult male goats were randomly divided into experimental group(n = 7)and control group(n = 7).All goats in the experimental group underwent conventional surgical thoracotomy under general anesthesia.During the operation,pacing electrodes were implanted in the left and right auricles.The AF model was established by continuous pacing at a fixed stimulation frequency of 600 times / minute for 4 weeks.The control group received the same operation without pacing.2.Observe the effective refractory period of atrium,and observe whether there is fibrosis in left and right atrium,left ventricle and vestibular tissue of pulmonary vein with Masson staining.3.Detection of m RNA expression by deep sequencingThe total RNAs were extracted from the left auricle of 11 goats(AF group 4,control group 7).The expression of m RNA was detected by RNA SEQ sequencing.4.Using quantitative real-time PCR(QRT PCR)to verify the accuracy of sequencing results.5.Bioinformatics analysisGSEA enrichment analysis of all transcriptional lines was performed to explore pathways related to AF remodeling.Result1.Successful establishment of epicardial rapid pacing model of atrial fibrillation in goats,4 successful,3 dead,4 successful goats can induce atrial fibrillation.The effective refractory period(AERP)was significantly shortened in AF group.2.The results of mass staining showed that there was no significant difference in the degree of fibrosis between the AF group and the control group.The fibrosis degree of pulmonary vein vestibule,bilateral heart and left ventricle decreased gradually in the two groups.3.It was found that the expression of RNAs in the left atrial appendage of goats in AF group was significantly different from that in sinolv group.The expression of genes such as enchig0000018618,enchig000023563,enchig000021270,enchig0000007244,enchig000023417 was significantly down regulated,and enchig0000017433,enchig000024130,enchig0000006316,enchig000012791,enchig000017462 were significantly down regulated Because the expression was obviously down regulated.Three RNAs were randomly selected for QRT PCR.The results confirmed the reliability of the sequencing results.4.GSEA analysis showed that the differentially expressed genes were mainly involved in inflammatory response,collagen metabolism,oxidative phosphorylation and other cellular pathways.conclusion1.AERP was significantly shortened in AF group.2.The results of in-depth sequencing showed that there was significant difference in m RNA expression between the two groups,and the results of QRT PCR confirmed that the sequencing results were true and reliable.3.GSEA analysis showed that there were significant differences in the expression of genes related to cell signaling pathways,such as inflammatory response,collagen fiber metabolism,oxidative phosphorylation,suggesting that inflammation and fibrosis play an important role in the myocardial remodeling of atrial fibrillation. |
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