丘脑束旁核lncRNA在心肌缺血再灌注损伤中的作用及其干预研究

There is obvious progress in understanding the mechanisms of brain-heart crosstalk. Myocardial ischemia-reperfusion (MI/RI) may lead to severe circulatory sequela, and even cause direct mortality of patients. It has been reported that thalamic parafascicular nucleus (Pf) modifies cardiac autonomic nervous system activity changes. Our previous researchers have found that the expression of Pf lncRNA was increased after MI/RI, which was involved in the reperfusion injury-related inflammatory response. Therefore, this project raises a presumption: abnormal expression of specific lncRNA in Pf may play a key role in the control of MI/RI. In the present study, by establishing a rat model of myocardial ischemia-reperfusion injury, we observe NONRATT025386 expression in the Pf and its associated injuries. We then inhibit Pf NONRATT025386 expression by using NONRATT025386-siRNA to investigate potential protective effects on the myocardial cells and related signaling pathways. Furthermore, we try to understand the metabolic mechanisms of Pf LncRNAs participating in the MI/RI by quantitative proteomics and 1HNMR metabolomics. This study offers the possibility to develop a novel therapeutic strategy to specifically attenuate MI/RI.

脑-心稳态“对话”机制的研究正逐步深入。心肌缺血再灌注损伤可引起循环系统严重后遗症直接威胁病人生命。然已有报道丘脑束旁核可以调控心脏自主神经节活性变化。申请者前期研究发现心肌缺血再灌注损伤，通过支配心脏的神经引起丘脑束旁核lncRNAs的表达变化。因此提出假设：丘脑束旁核特异性lncRNAs表达异常在心肌缺血再灌注损伤的调控中起重要作用。本项目拟建立大鼠结扎冠状动脉诱发局部心肌缺血导致整体心肌损伤模型，给予局部沉默束状核lncRNAs，观察下调束状核NONRATT025386对心肌损伤的保护作用及机制，并运用定量蛋白质组与代谢组学技术阐明束状核lncRNAs参与心肌缺血再灌注损伤中的代谢机制，为特异性减轻心肌缺血再灌注损伤提供一种新的干预策略。

脑-心稳态“对话”机制的研究正逐步深入。本项目是研究丘脑束旁核特异性lncRNAs表达异常在心肌缺血再灌注损伤的调控中起重要作用。首先构建腺相关病毒AAV- NONRATT025386-siRNA-GFP 并进行功能评价；之后将AAV-NONRATT025386-siRNA-GFP和相关空病毒立体定向注射病毒至丘脑束旁核。将SD大鼠分为空白对照组、空病毒对照组、模型组(MIRI组)和4组不同剂量AAV-NONRATT025386-siRNA模型组。观察下调束状核NONRATT025386对心肌损伤的保护作用及机制，并运用定量蛋白质组与代谢组学技术阐明束状核lncRNAs参与心肌缺血再灌注损伤中的代谢机制。超声心动图评估结果显示心脏缺血再灌注导致心功能不全。与对照组相比，MIRI组代谢物改变为牛磺酸和胆碱，差异主要发生在丘脑和脑干。与对照组相比，MIRI组血清肌钙蛋白水平明显升高，且心肌TTC染色发现梗死心肌组织，HE染色显示MIRI组和低剂量AAV-siRNA组心肌细胞排列紊乱，可见炎症细胞浸润，而高剂量AAV-siRNA组上述指标则明显好于MIRI组和低剂量AAV-siRNA组。NMR代谢组学分析发现与相比，MIRI组丘脑牛磺酸含量高于对照组(33.4%↑, P=0.032), 胆碱含量高于对照组(40.5%↑, P=0.010)。牛磺酸和胆碱可能参与束状核lncRNAs对心肌缺血再灌注损伤的调控。此研究为特异性减轻心肌缺血再灌注损伤提供一种新的干预策略。