To Analyze The MiRNA And LncRNA Related To Notch1 Signal Pathway In The Myocardial Protection Of Autophagy

Chapter 1 IntroductionAcute myocardial infarction(AMI) has an extremely high morbidity and mortality rate in cardiovascular disease, ischemia/reperfusion injury(IRI) is still a difficult problem faced to the clinic treatment of AMI. How to alleviate IRI is the research hotspot of myocardial injury and protection. Research on the mechanism of myocardial injury, which can provide the theory and practice basis of the clinical prevention and treatment of myocardial ischemia injury. Therefore, we based on the lnc RNA-mi RNA-Notch1 regulation hypothesis, established the hypoxia/reoxygenation(H/R) injury and autophagy protection models in myocardial cell, using high-throughput sequencing and bioinformatics analysis to analyze the mi RNA of regulating Notch1 protein in autophagy protection and the lnc RNA of regulating these mi RNAs. In order to provide the scientific basis for the molecule mechanism of Notch1 signal pathway in the myocardial protection of autophagy. Chapter 2 Notch1 signal pathway enhanced the myocardial protection of autophagy against hypoxia-reoxygenation injuryObjective: To analyze the reagent concentration of 3-MA and RAPA for effectively regulate the autophagy, and discuss the role of Notch1 signaling pathway in the myocardial protection of autophagy against hypoxia-reoxygenation(H/R) injury.Methods: We established the H/R injury model in myocardial cell, provided 3-MA and RAPA in the model, then detected the cell vitality, analyzed the effective reagent concentration of 3-MA and RAPA. Using Ad-N1 ICD and Ad-N1ICD-sh RNA to effect Notch1 signaling pathway, then detected the cell vitality.Results: In the study, the effective reagent concentration of 3-MA and RAPA was 1μM and 50 n M respectively. Overexpression Notch1 could increase the cell vitality after H/R(p<0.01), interference Notch1 could decrease the cell vitality after H/R(p<0.01), 3-MA and overexpression Notch1 could decrease the cell vitality after H/R(p<0.01), RAPA and interference Notch1 could recover the cell vitality after H/R(p<0.01).Conclusions: Notch1 signal pathway enhanced the myocardial protection of autophagy against H/R injury.Chapter 3 To analyze the mi RNA and lnc RNA related to Notch1 signal pathway in the myocardial protection of autophagy by the high-throughput sequencing and bioinformatics comparative analysisObjective: Using the high-throughput sequencing and bioinformatics comparative analysis to analyze the mi RNA and lnc RNA, which regulate Notch1 signal pathway in the myocardial protection of autophagy.Methods: Extracted the total RNA from H/R injury model in myocardial cell, used the high-throughput sequencing and bioinformatics analysis to analyze the differential expression mi RNA and lnc RNA.Results: We found that mi R-702-5p、mi R-344g、mi R-323-5p、mi R-6334、mi R-5132-5p、mi R-15b-5p and mi R-3562 could regulate the gene expression of Notch1. Bioinformatics analysis showed that lnc RNA-4321 could directly bond to mi R-702-5p and effect its activity. The expression of lnc RAN-4321 and mi R-702-5p in cardiomyocytes treated with RAPA or 3-MA were detected by Realtime-PCR, the results showed that the expression of lnc RNA-4321 was proportional to autophagy, but the expression of mi R-702-5p was inversely proportional to autophagy.Conclusions: lnc RNA-4321/mi R-702-5p/Notch1 may play an important role in the myocardial protection of autophagy against IRI.