lncRNA-MIRA1433/miR-375/14-3-3η对心肌I/R损伤不同阶段自噬的双重调节

在总结前人与自身工作基础上，我们假设：心肌急性缺血/再灌注（I/R）的不同阶段，lncRNA-MIRA1433/miR-375/14-3-3η信号通路相互整合、双重调节自噬、减轻心肌细胞损伤。拟将lncRNA-MIRA1433/miR-375/14-3-3η有机结合，采用荧光“自噬流”动态检测，在确认14-3-3η对H9C2心肌样细胞及其转基因细胞双重调节自噬之基础上，运用RIP、RNA Pull-Down、双荧光素酶报告基因技术等，建立相应的转基因细胞，观察不同lncRNA-MIRA1433/miR-375/14-3-3η表达水平、彼此间相互作用对自噬的调控，以及对其抗急性I/R损伤能力的影响；最后在整体动物实验上比较论证。这将有助于深入、全面了解心肌急性I/R损伤现象及其细胞自噬之作用，完善、丰富相关知识和理论，阐明可能新的药物作用之靶点，为心肌保护药物的研发奠定良好的理论与实验基础。

In light of previous work by us and others, we hypothesize that at different stage of acute myocardial ischemia/reperfusion(I/R), lncRNA-MIRA1433/miR-375 /14-3-3ηsignal pathway could alleviate the myocardial injury through mutual integration and dual regulative effect on autophagy. We plan to combine the lncRNA-MIRA1433/miR-375/14-3-3 pathway and use the fluorescence "autophagy flow” dynamic detection to recognize the dual regulative effect on autophagy of 14-3-3η on H9C2 cardiomyocyte like cells and it’s transgenic cells.Based on this, we plan to use the RIP,RNA pull-down, double luciferase reporter gene technology to establish the corresponding transgenic cells,then observe the different expression level of lncRNA-MIRA1433/miR-375/14-3-3η, it’s regulative effect on autophagy through interaction with each other and it’s anti acute I / R injury effects, finally compare and comfirm the results through whole animal experiment. The project will help us deeply and comprehensively understand the acute myocardial I/ R injury phenomenon and it’s cell autophagy, improve and enrich the relevant knowledge and theory, clarify the potential target of new drugs, lay a good theoretical and experimental foundation for the research and development of myocardial protective drugs.