Remote Ischemic Postconditioning Inhibited Mitophagy to Achieve Neuroprotective Effects in the Rat Model of Cardiac Arrest
Remote Ischemic Postconditioning Inhibited Mitophagy to Achieve Neuroprotective Effects in the Rat Model of Cardiac Arrest
复制标题
远程缺血后处理抑制线粒体自噬，在心脏骤停大鼠模型中实现神经保护作用
DOI：
10.1007/s11064-020-03193-x
复制
发表时间：
2021-01-06
影响因子：
4.4
通讯作者：
Zhu, ShuiBo
中科院分区：
文献类型：
--
作者：
Huang, Yang;Gao, Xuhui;Zhu, ShuiBo
远程缺血后处理（RI-postC）是改善心脏骤停后神经功能的有效措施。然而，RI-postC的脑保护机制尚未完全阐明，是否与线粒体自噬有关尚不清楚。本研究利用大鼠心脏骤停模型，研究RI-postC对线粒体自噬的影响，并探讨其可能的信号通路。将大鼠随机分为Sham组、CA/CPR组、Mdivi-1组和RI-postC组。采用窒息法建立心脏骤停动物模型。通过夹闭和松开左股动脉进行RI-postC。Mdivi-1单次静脉注射治疗。免疫印迹法检测TOMM 20、TIM 23、Mfn 1、PINK 1和parkin的表达。流式细胞仪测定线粒体膜电位。实时荧光定量PCR检测线粒体DNA相对含量。流式细胞术和TUNEL法检测海马神经元凋亡。此外，还进行了组织学检查。结果显示，RI-postC与线粒体自噬抑制剂Mdivi-1相似，可抑制心肺复苏后线粒体自噬相关蛋白水平的下降，提高线粒体膜电位，上调mt-Atp 6/Rp 113比值。此外，RI-postC还能降低CPR后海马神经元凋亡率，减轻CPR后海马神经元的损伤。此外，RI-postC和Mdivi-1可降低CPR后线粒体中PINK 1和parkin的蛋白水平，而增加胞浆中PINK 1的水平。这些结果表明，RI-postC可以通过PINK 1/parkin信号通路抑制过度激活的线粒体自噬，从而提供神经保护作用。
Remote ischemic postconditioning (RI-postC) is an effective measure to improve nerve function after cardiac arrest. However, the brain protective mechanism of RI-postC has not been fully elucidated, and whether it is related to mitophagy is unclear. In this study, we used the rat model of cardiac arrest to study the effect of RI-postC on mitophagy and explore its possible signaling pathways. Rats were randomly divided into Sham group, CA/CPR group, Mdivi-1 group and RI-postC group. The animal model of cardiac arrest was established by asphyxia. RI-postC was performed by clamping and loosening the left femoral artery. Mdivi-1 was treated with a single intravenous injection. Levels of TOMM20, TIM23, Mfn1, PINK1 and parkin were detected by western blots. Mitochondrial membrane potential was measured by flow cytometry. Real-time PCR was used to detect relative mitochondrial DNA levels. The apoptosis of hippocampal neurons was detected by flow and TUNEL. In addition, Histopathological tests were performed. The results showed that RI-postC was similar to the mitophagy inhibitor Mdivi-1, which could inhibit the decrease of mitophagy-related protein level, improve mitochondrial membrane potential and up-regulate the ratio of mt-Atp6/Rpl13 after cardiopulmonary resuscitation (CPR). Furthermore, RI-postC could also reduce the rate of hippocampal nerve apoptosis and the damage of hippocampal neurons after CPR. Moreover, RI-postC and Mdivi-1 could reduce the protein levels of PINK1 and parkin in mitochondria after CPR, while increasing PINK1 levels in the cytoplasm. These findings suggested that RI-postC could inhibit the overactivation mitophagy through the PINK1/parkin signaling pathway, thus providing neuroprotective effects.