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W026B是一种对小鼠脑缺血再灌注(I-R)损伤具有保护作用的新化合物,但其具体的作用机制尚不明确.本研究应用蛋白质组学观察了W026B在I-R损伤脑组织中对蛋白表达的影响,并试图揭示其潜在的靶点.通过蛋白质组学共检测到4852种蛋白,在脑I-R模型组和W026B治疗组中发现了42种发生显著变化的蛋白,这些蛋白大多与免疫炎症、代谢、神经保护以及细胞增殖和细胞结构有关.经Western blotting验证,在五种候选蛋白中有三种蛋白表现出与蛋白质组学检测结果一致的改变.我们选择RGS17作为进一步研究的对象,利用siRNA RGS 17敲低RGS17蛋白后加重了脑损伤,并抵消了W026B的保护作用.W026B可与RGS17结合(KD:6.04×10-6 mol/L).RGS17的下调加重了Ⅰ型代谢型谷氨酸受体激动剂(Ⅰ型mGluRs)诱导的Neuro-2a细胞损伤,并抵消了W026B的保护作用.综上所述,W026B通过影响多种蛋白保护大脑免受I-R损伤,RGS17可能是W026B的靶点之一,也是潜在的脑I-R损伤治疗靶点.受W026B影响的RGS 17调节的G蛋白上游受体可能是Ⅰ型mGluRs.本研究为W026B的进一步研发和潜在的临床应用提供了有益的证据.“,”W026B is a new compound that has a protective effect on cerebral ischemia reperfusion (I-R) injury in mice,while its specific mechanism is still unknown.In this study,proteomics was used to observe the effect of W026B on protein expression in brain I-R tissue,and to reveal its potential target.A total of 42 significantly altered proteins were identified in both brain I-R model and W026B treatment from 4852 proteins detected by proteomics,and most of these proteins were related to immunity and inflammation,metabolism,neuroprotection as well as cell proliferation and cell structure.Western blotting analysis showed that three out of five selected proteins showed consistent alteration with the proteomics.Regulator of G protein signaling 17 (RGS 17) was selected for further study,and its knockdown by siRNA RGS17 aggravated brain injury and abolished the protective effect of W026B.W026B could bind with RGS 17 (KD:6.04× 10-6 mol/L).The knockdown of RGS 17 aggravated Neuro-2a cell damage induced by group I metabotropic glutamate receptors (mGluRs) agonist,and abolished the protective effect of W026B.In conclusion,W026B protected brain against I-R injury by affecting diverse proteins.RGS17 might be one of its targets and a potential therapeutic target of brain I-R injury.The upstream receptor of G protein,which was regulated by RGS 17 and affected by W026B,might be group Ⅰ mGluRs.This study provided useful evidence for the further R&D and the potential clinical application of W026B.