ShcA及相关的长链非编码RNA在心肺复苏后脑损伤中的作用及机制研究

Post-cardiac arrest (CA) and cardiopulmonary resuscitation (CPR) cerebral injury is the main reason of death of patients underwent successful CPR. The specific mechanisms of post CA/CPR cerebral injury and potential neuroprotective target need to be clarified. In our previous work, we analyzed the information of mRNA and long non-coding RNA (lncRNA) sequencing and confirmed the up-regulated gene named ShcA and related lncRNA named lncRNA-PS in the hippocampus of mice underwent CA/CPR. Additionally, we found that down-regulation of ShcA through perioperative intracerebroventricular injection of ShcA siRNA could significant improve the neurological outcome in mice underwent CA/CPR. This indicates that ShcA and related lncRNA may involve in post CA/CPR cerebral injury. We design this research to further confirm the relationship between the expression of ShcA and lncRNA-PS and post CA/CPR cerebral injury in molecular, cellular and tissue levels. We will also investigate the possible mechanisms for the development of post CA/CPR cerebral injury mediated by ShcA and lncRNA-PS. In addition, we will clarify means of regulation of ShcA by lncRNA-PS and related molecular mechanisms will be determined in our designed experiments. Moreover, clinical study regarding the role of ShcA and associated lncRNA in the development of post CA/CPR cerebral injury will be conducted. This research will provide novel view and theoretical foundation of mechanisms of post CA/CPR cerebral injury and an attractive therapeutic target.

心跳骤停(CA)及心肺复苏(CPR)后脑损伤被认为是CPR成功后患者死亡的最主要原因，亟需明确其发病机制及潜在治疗靶点。我们前期通过mRNA和长链非编码RNA (lncRNA)测序的信息采集分析，初步确定了CPR组小鼠海马组织中表达上调的ShcA及相关的lncRNA，命名为lncRNA-PS；同时下调ShcA (siRNA干扰)可显著改善小鼠CPR后的神经功能，这提示ShcA及相关lncRNA可能参与介导了CPR后脑损伤的发生。本课题拟进一步从分子、细胞和整体不同层面，明确CA/CPR后引起ShcA和lncRNA-PS的表达变化及其与CPR后脑损伤的关系，探讨ShcA和lncRNA-PS介导CPR后脑损伤发生的可能机制，阐明lncRNA-PS调控ShcA的作用方式及分子机制，并进行临床探索性研究。本项目的成功实施将为CPR后脑损伤的发病机制及防治措施提供新的思路和理论基础。

心跳骤停(CA)及心肺复苏(CPR)后脑损伤被认为是CPR成功后患者死亡的最主要原因，亟需明确其发病机制及潜在治疗靶点。我们通过mRNA和长链非编码RNA (lncRNA)测序的信息采集分析，确定了CPR组小鼠海马组织中表达上调的ShcA及相关的lncRNA-PS；同时下调ShcA (siRNA干扰)可显著改善小鼠CPR后的神经功能，这提示ShcA及相关lncRNA可能参与介导了CPR后脑损伤的发生。我们进一步通过Cre/LoxP条件性基因敲除实现小鼠海马CA1区神经元中ShcA的抑制。体外实验中，我们使用针对lncRNA-PS和蛋白ShcA的慢病毒实现对HT22细胞系的基因调控。自主循环(ROSC)恢复后的小鼠存活率和神经功能评分明显下降，认知功能受到影响。通过原位杂交和免疫荧光技术，我们发现LncRNA-PS和蛋白ShcA在CA/CPR后的过表达主要发生在海马CA1区神经元中。CA/CPR前敲除小鼠海马CA1区神经元中的ShcA可减轻CA/CPR后的脑损伤。ShcA敲除对HT22细胞系OGD/R之后的保护性作用是通过抑制炎症和细胞凋亡实现的，而lncRNA-PS的过表达逆转了这一作用。