Peroxynitrite (ONOO⁻)-mediated mitophagy activation represents a vital pathogenic mechanism in ischemic stroke. Our previous study suggests that ONOO⁻ mediates Drp1 recruitment to the damaged mitochondria for excessive mitophagy, aggravating cerebral ischemia/reperfusion injury and the ONOO⁻-mediated mitophagy activation could be a crucial therapeutic target for improving outcome of ischemic stroke. In the present study, we tested the neuroprotective effects of rehmapicroside, a natural compound from a medicinal plant, on inhibiting ONOO⁻-mediated mitophagy activation, attenuating infarct size and improving neurological functions by using the in vitro cultured PC12 cells exposed to oxygen glucose deprivation with reoxygenation (OGD/RO) condition and the in vivo rat model of middle cerebral artery occlusion (MCAO) for 2 h of transient cerebral ischemia plus 22 h of reperfusion. The major discoveries include following aspects: (1) Rehmapicroside reacted with ONOO⁻ directly to scavenge ONOO⁻; (2) Rehmapicroside decreased O₂⁻ and ONOO⁻, up-regulated Bcl-2 but down-regulated Bax, Caspase-3 and cleaved Caspase-3, and down-regulated PINK1, Parkin, p62 and the ratio of LC3-II to LC3-I in the OGD/RO-treated PC12 cells; (3) Rehmapicroside suppressed 3-nitrotyrosine formation, Drp1 nitration as well as NADPH oxidases and iNOS expression in the ischemia-reperfused rat brains; (4) Rehmapicroside prevented the translocations of PINK1, Parkin and Drp1 into the mitochondria for mitophagy activation in the ischemia-reperfused rat brains; (5) Rehmapicroside ameliorated infarct sizes and improved neurological deficit scores in the rats with transient MCAO cerebral ischemia. Taken together, rehmapicroside could be a potential drug candidate against cerebral ischemia-reperfusion injury, and its neuroprotective mechanisms could be attributed to inhibiting the ONOO⁻-mediated mitophagy activation.

过氧亚硝酸盐（ONOO ^- ）介导的自噬激活表示在缺血性中风的重要致病机制。我们以前的研究表明，ONOO ^-介导DRP1招聘过多的自噬受损的线粒体，加重脑缺血/再灌注损伤和ONOO ^-介导的自噬激活可能是改善缺血性中风的结果的关键治疗目标。在本研究中，我们测试rehmapicroside的神经保护作用，从药用植物，在抑制ONOO的天然化合物^-通过使用介导的自噬活化，减弱梗塞面积和改善神经功能的体外培养的PC12细胞暴露于缺氧复氧（OGD / RO）条件下，并在体内大鼠大脑中动脉闭塞（MCAO）模型中进行了2 h的短暂性脑缺血再灌注22 h。的重大发现包括以下几个方面：（1）与Rehmapicroside ONOO反应^-直接向扫气ONOO ^- ; （2）降低RehmapicrosideÒ ₂ ^-和ONOO ^-OGD / RO处理的PC12中的Bcl-2，Bcl-2上调但Bax，Caspase-3和Caspase-3裂解下调，PINK1，Parkin，p62以及LC3-II与LC3-I的比例下调细胞; （3）Rehmapicroside抑制了缺血再灌注大鼠脑中的3-硝基酪氨酸形成，Drp1硝化以及NADPH氧化酶和iNOS的表达；（4）Rehmapicroside阻止了PINK1，Parkin和Drp1易位进入线粒体，从而在缺血再灌注的大鼠脑中激活了线粒体。（5）Rehmapicroside改善了短暂MCAO脑缺血大鼠的梗死面积并改善了其神经功能缺损评分。总之，rehmapicroside可能是对脑缺血再灌注损伤的潜在的候选药物，其神经保护机制，可以归因于抑制ONOO ^-介导的线粒体激活。