ciRS-7/miR-7/mTOR/p70S6K信号通路介导神经元自噬导致大鼠复苏后脑损伤的机制及参附注射液的干预研究

Cerebral ischemia-reperfusion injury after cardiac arrest is the key pathological process of cerebral resuscitation after cardiopulmonary resuscitation. Recent studies show that neuronal autophagy is a key mechanism of ischemic neuronal damage. In particular, a large number of circular RNAs (ciRS-7) exist in the nervous system and lead neuron to autophagy by affecting expression of miRNA-7 and then to interfere expression of mTOR and it's downstream target p70S6K. Our previous studies find that Shenfu injection can block the neurological function of brain injury after cardiopulmonary resuscitation by blocking Nogo1-NgR, suggests that Shenfu injection can promote the repair function of neuronal cells. No reports about neuronal autophagy on the mechanism of brain injury after cardiopulmonary resuscitation have been reported. Based on these, we hypothesize that Shenfu injection may protect brain cells by regulating the ciRS-7 / miR-7 / mTOR / p70S6K signaling pathway to mediate neuronal autophagy after CPR. In this study, the ciRS-7 / miR-7 / mTOR / p70S6K signaling pathway will be detected by immunohistochemistry, weston-blot, electron microscopy and immunofluorescence confocal microscopy on duplicated rat models of cardiopulmonary resuscitation (CPR), to explore the mechanism of neuronal autophagy after cardiopulmonary resuscitation and the intervention effect of Shenfu injection, reveal the effective therapeutic target of Shenfu injection on cerebral ischemia-reperfusion injury after cardiopulmonary resuscitation, and provide a new objective basis for the treatment of cerebral injury after resuscitation by Shenfu Injection.

心脏骤停后脑部缺血再灌注损伤是心肺复苏后脑复苏的关键病理环节。最新研究表明，神经元自噬是缺血性神经元损伤的关键机制。大量的环状RNA （ciRS-7）存在神经系统，并通过miRNA-7表达影响mTOR及下游靶物-p70S6K导致神经元自噬。我们前期研究也发现，参附注射液通过阻断Nogo1-NgR对神经元细胞修复功能有促进作用。目前未见关于神经元自噬对心肺复苏后脑损伤机制的研究报道，据此我们提出假说：参附注射液可能通过调节ciRS-7/miR-7/mTOR/p70S6K信号通路介导心肺复苏后神经元自噬而起到脑保护作用。本课题通过复制心肺复苏后大鼠模型，用免疫组织化学法等检测ciRS-7/miR-7/mTOR/p70S6K信号通路；探讨心肺复苏后神经元自噬的机制和参附注射液的干预效应，揭示参附注射液对心肺复苏后脑部缺血再灌注损伤的有效治疗靶点，为参附注射液治疗复苏后脑损伤提供新的客观依据。

心脏骤停后脑缺血再灌注损伤是心肺复苏后脑损伤的重要病理环节。神经元细胞自噬是缺血性神经损伤的关键机制。大量的环状RNA（ciRS-7）存在神经系统，并通过miRNA-7表达影响mTOR及下游靶物-p70S6K导致神经元自噬。我们前期研究发现,参附注射液通过阻断勿动蛋白受体（Nogo1-NgR）对神经元细胞修复功能有促进作用。但未见神经元细胞自噬对心肺复苏后脑损伤机制的研究报道，据此我们提出假说：参附注射液可能通过调节ciRS-7/miR-7/mTOR/p70S6K信号通路介导心肺复苏后神经元自噬而起到脑保护作用。本课题通过复制心脏骤停/心肺复苏后小鼠模型，用qPCR、TUNEL荧光、透射电镜、免疫组化和尼氏（NISSL）染色等检测ciRS-7/miR-7/mTOR/p70S6K信号通路； 最后得出：.1.参附注射液能提高心脏骤停/心肺复苏（CA/CPR）小鼠的生存率和降低神经功能严重程度评分，对脑损伤有神经保护作用。.2.通过复制心脏骤停/心肺复苏（CA/CPR）所致心脏骤停后脑损伤的小鼠模型，脑部缺血再灌注可能下调ciRS-7的表达，进而对其下游的miR-7抑制作用减弱，而上调miR-7/mTOR/p70S6K信号通路的蛋白表达，导致神经细胞自噬和凋亡的发生。.3.参附注射液可能上调ciRs-7的表达，对miR-7抑制作用增强，下调下游的miR-7/mTOR/p70S6K蛋白表达，从而减少心肺复苏后小鼠神经细胞的自噬和凋亡的发生，发挥神经细胞的保护作用。探讨心肺复苏自主循环恢复后神经元细胞自噬的机制和参附注射液的干预效应，揭示参附注射液对心肺复苏后脑缺血再灌注损伤的有效治疗靶点，为参附注射液治疗心肺复苏后脑损伤提供新的理论依据。