Study On The Mechanism Of Paeonol Combined With Paeoniflrinon Acute Myocardial Ischemia Injury Based On Microrna Regulation Pathway

Objective: We use paeonol combined with paeoniflorin to treat the rats with acute myocardial ischemia injury,the miRNA of different expression in heart tissue of rats with acute myocardial injury was used as the entry point of the study,and further discusses the mechanism of miRNA regulating the effect of paeonol combined with paeoniflorin on myocardial ischemia injury.Methods: 1.A rat model of acute myocardial ischemia was established,and the intervention of paeonol combined with paeoniflorin was carried out.The differences between microRNA in the myocardium of the model group and the paeonol combined with paeoniflorin group were screened by gene chip,and then verified by RT-PCR.2.Bioinformatics analysis of differentially expressed miRNA.We constructed the regulatory network between miRNA and Gene,GO and Pathway,and got the key up regulated miRNA and down regulated miRNA,and the main signaling pathways.3.Construction of rat acute myocardial damage model,the corresponding intervention of paeonol combined with paeoniflorin compatibility,further verified by bioinformatics prediction results of experimental monitoring ECG,myocardial infarction area was measured and the protein concentration determination,immune response,explored the regulation of miRNA network in paeonol combined with paeoniflorin myocardial ischemia injury protection mechanism.4.The hypoxic model of rat H9c2 cells was established.The intervention of paeonol combined with paeoniflorin,miRNA-21 analogue,miRNA-21 inhibitor and p38 MAPK signal pathway inhibitor(SB203580)were carried out,the level of TNF-? was detected by ELISA,and the MAPK signal pathway was detected by Western blot,in order to explore the mechanism of myocardial ischemia regulated by paeonol combined with paeoniflorin.Results: 1.When myocardial ischemia injury was treated by paeonol combined with paeoniflorin,the key expression of myocardial differential expression was Up-regulatedmiRNA-141-3p and down-regulated miRNA-21-5p,miRNA-92a-3p and miRNA-223-3p.2.The most critical up-regulated miRNA in the regulation network is miRNA-141-3p,and the most critical down regulation of miRNA is miRNA-21-5p.Both of them regulate the MAPK signaling pathway,and the correlation between the two regulators is the highest.3.The combination of paeonol and paeoniflorin has an obvious inhibitory effect on S-T segment elevation,infarct size,the activity of CK,LDH and MDA content in acute myocardial ischemia,elevate SOD activity,inhibit the expression of p38 and JNK signaling pathway,and promote the activity of ERK signal pathway.4.In H9c2 cells,paeonol combined with paeoniflorin further inhibits the secretion of TNF-a by regulating the expression level of miRNA-21;paeonol combined with paeoniflorin can inhibit the activation of p38 MAPK signaling pathway;meanwhile,paeonol combined with paeoniflorin can reverse the activation of miRNA-21 analog p38 MAPK signaling pathway;paeonol combined with paeoniflorin and miRNA-21 inhibitors can activate p38 MAPK.Conclusion: 1.The most critical up-regulation of miRNA is the miRNA-141-3p.The most critical down-regulation of miRNA is the miRNA-21-5p and the MAPKs is an important signaling pathway.2.The compatibility of paeonol and paeoniflorin can promote the activity of ERK signaling pathway by inhibiting the p38 and JNK signaling pathways,and play an important role in protecting the myocardium.3.The combination of paeonol and paeoniflorin can inhibit the expression of miRNA-21,inhibit the p38 MAPK signaling pathway,inhibit the secretion and release of important inflammatory factors during myocardial ischemia,and then play a protective role in the ischemic myocardium.