Restoration of early deficiency of axonal guidance signaling by guanxinning injection as a novel therapeutic option for acute ischemic stroke
Restoration of early deficiency of axonal guidance signaling by guanxinning injection as a novel therapeutic option for acute ischemic stroke
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冠心宁注射液恢复早期轴突引导信号传导缺陷作为急性缺血性脑卒中的新治疗选择
DOI：
10.1016/j.phrs.2021.105460
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发表时间：
2021
影响因子：
9.3
通讯作者：
Zhu Yan
中科院分区：
文献类型：
--
作者：
Xiao Guangxu;Lyu Ming;Li Zhixiong;Cao Linghua;Liu Xinyan;Wang Yule;He Shuang;Chen Zihao;Du Hongxia;Feng Yuxin;Wang Jigang;Zhu Yan
尽管它的发病率和死亡率很高，但由于我们对其发病机制的分子机制了解不完全，目前仍缺乏有效的治疗方法。在这项研究中，我们证明了SHH-ptch1-GLI1介导的轴突引导信号及其相关的神经发生，作为神经元发育的中心途径，在急性卒中模型的早期也发挥了关键作用。特别是在体内，我们通过大脑中动脉栓塞法评价了GXNI对缺血性中风小鼠的影响，发现GXNI通过减少脑梗塞体积、神经功能评分和脑水肿，逆转血脑屏障通透性和组织病理学改变，显著减轻脑缺血再灌注(I/R)损伤。采用RNA-SEQ和网络药理学分析相结合的方法，结合RT-PCR、免疫组织化学和Western blotting验证GXNI的作用机制。指出轴突引导信号通路在GXNI中的作用最为显著，Shh、ptch1和Gli1基因在脑保护中起关键作用。此外，GXNI还能明显保护原代培养的大脑皮层神经元细胞免受缺氧缺糖/复氧损伤，并促进受损神经元的轴突生长和突触形成，进一步证实了活体实验的结果。此外，由于环丙胺抑制SHH-ptch1-GLI1信号通路，GXNI的作用明显减弱。因此，我们的研究通过SHH-ptch1-GLI1介导的轴突引导信号为临床应用GXNI治疗急性缺血性卒中提供了新的选择，轴突指导信号是先前被认为是卒中后恢复的神经元发育途径。
Despite of its high morbidity and mortality, there is still a lack of effective treatment for ischemic stroke in part due to our incomplete understanding of molecular mechanisms of its pathogenesis. In this study, we demonstrate that SHH-PTCH1-GLI1-mediated axonal guidance signaling and its related neurogenesis, a central pathway for neuronal development, also plays a critical role in early stage of an acute stroke model. Specifically,in vivo, we evaluated the effect of GXNI on ischemic stroke mice via using the middle cerebral artery embolization model, and found that GXNI significantly alleviated cerebral ischemic reperfusion (I/R) injury by reducing the volume of cerebral infarction, neurological deficit score and cerebral edema, reversing the BBB permeability and histopathological changes. A combined approach of RNA-seq and network pharmacology analysis was used to reveal the underlying mechanisms of GXNI followed by RT-PCR, immunohistochemistry and western blotting validation. It was pointed out that axon guidance signaling pathway played the most prominent role in GXNI action withShh,Ptch1, andGli1genes as the critical contributors in brain protection. In addition, GXNI markedly prevented primary cortical neuron cells from oxygen-glucose deprivation/reoxygenation damagein vitro, and promoted axon growth and synaptogenesis of damaged neurons, which further confirmed the results ofin vivoexperiments. Moreover, due to the inhibition of the SHH-PTCH1-GLI1 signaling pathway by cyclopropylamine, the effect of GXNI was significantly weakened. Hence, our study provides a novel option for the clinical treatment of acute ischemic stroke by GXNI via SHH-PTCH1-GLI1-mediated axonal guidance signaling, a neuronal development pathway previously considered for after-stroke recovery.