| Research background:Myocardial ischemia-reperfusion injury(I/R),with a very high fatality rate,is a cardiac pathological change caused by secondary ischemia or occlusion of coronary arteries after events such as stroke,coronary heart disease,and organ transplantation.Therefore,it is very important to explore the mechanism of myocardial ischemia-reperfusion injury and new strategies for prevention and treatment.In recent years,studies have found that autophagy and ferroptosis play a core role in the pathogenesis and progression of ischemia-reperfusion injury,but the mechanism is still unclear.Therefore,exploring the homeostasis of autophagy and ferroptosis is of great significance for regulating the process of myocardial ischemia-reperfusion injury.As an RNA-binding protein,ELAVL1 could participate in the post-transcriptional regulation of gene expression.Study shows that ELAVL1 is involved in the autophagy-ferroptosis process in liver fibrosis,and another research has shown that ELAVL1 is up-regulated in myocardial ischemia-reperfusion injury and is involved in the regulation of cardiomyocytes apoptosis process.The above information suggests that in the pathogenesis of myocardial ischemia-reperfusion injury,ELAVL1 may play a regulatory role through the autophagy-ferroptosis pathway,which is worthy of further exploration.Main research and experimental methods:Mouse I/R model and in vitro cardiomyocyte hypoxia/reoxygenation(H/R)model were constructed,TCC staining was used to verify the degree of myocardial infarction in I/R model mice.Iron levels,LDH,GSH,GPx4,ROS and LPO activities were detected with the kit.qPCR and western blot were used to detect the expression of autophagy and ferroptosis-related genes.ChIP and dual-luciferase reporter gene experiments were used to verify the interaction between FOXC1 and the upstream promoter region of ELAVL1.Experimental results:①In the mouse I/R model,ELAVL was significantly up-regulated,the expressions of ferroptosis-related markers GPx4,FTH1,and GSH were down-regulated,and iron levels and LDH activity were significantly increased.②In the cardiomyocyte H/R model,ELAVL was significantly up-regulated,and knockdown of ELAVL 1 could significantly promote the expression of the negative regulator of ferroptosis GPx4 and inhibit the expression of Beclin-1 and LC3,reverse the H/R treatment-induced caused increased in ELAVL,Iron,ROS levels,LPO,LDH,and decreased in cell viability and GSH activity.③ In the cardiomyocyte H/R model,the autophagy inhibitor 3-MA significantly down-regulated Beclin-1,LC3II and up-regulated the expression of p62,and rescued H/R-induced cardiomyocyte viability,decreased GSH activity,and iron,ROS,and ROS levels,and increased LPO and LDH activity.④ In the mouse I/R model,down-regulation of ELAVL1 expression could inhibit up-regulation beclin-1 and LC3 expression and down-regulation of P62 and GPx4 expression induced by I/R,while activating autophagy(expressed BeClin-1)was able to reverse the above effect.⑤ ELAVL1 protein combined with Beclin-1 mRNA and improved the stability of Beclin-1 mRNA.FOXC1 protein combined with ELAVL1 promoter region and activated its expression.Conclusion:① ELAVL1 is up-regulated in myocardial ischemia-reperfusion injury,and myocardial ischemia-reperfusion injury promotes ferroptosis..②ELAVL 1 regulates myocardial ischemia-induced ferroptosis and promotes myocardial ischemia-reperfusion injury.③There is an interaction relationship between autophagy and ferroptosis.Autophagy can regulate the expression of key factors of ferroptosis and then mediate the process of myocardial ischemia-reperfusion injury.④ ELAVL1 regulates the process of myocardial ischemia-reperfusion injury by regulating autophagy-ferroptosis mediated by autophagy-related gene Beclin-1.⑤ FOXC1 is a key upstream mechanism of ELAVL1,which can positively regulate the transcription of ELAVL1,participate in the regulation of theautophagy-ferroptosis pathway,and is a key factor in myocardial ischemia-reperfusion injury. |
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