Department of Chemistry, Department of Physiology and Functional Genomics, Center for Research at the Bio/Nano Interface, UF Health Cancer Center , UF Genetics Institute and McKnight Brain Institute University of Florida , Gainesville , Florida 32611 , United States.
Shenzhen Research Institute , Hunan University , Shenzhen , Guangdong 518000 , China.
ACS Chem Neurosci. 2019 Jun 19;10(6):2877-2889. doi: 10.1021/acschemneuro.9b00103. Epub 2019 Apr 16.
Stroke is a leading cause of disability and the second leading cause of death among adults worldwide, while the mechanisms underlying neuronal death and dysfunction remain poorly understood. Here, we investigated the differential proteomic profiles of mouse brain homogenate with 3 h of middle cerebral artery occlusion (MCAO) ischemia, or sham, using Coomassie Brilliant Blue staining, followed by mass spectrometry. We identified enolase1 (ENO1), a key glycolytic enzyme, as a potential mediator of neuronal injury in MCAO ischemic model. Reverse transcription polymerase chain reaction and western blotting data showed that ENO1 was ubiquitously expressed in various tissues, distinct regions of brain, and different postnatal age. Immunohistochemical analysis revealed that ENO1 is localized in neuronal cytoplasm and dendrites. Interestingly, the expression level of ENO1 was significantly increased in the early stage, but dramatically decreased in the late stage, of cerebral ischemia in vivo. This dynamic change was consistent with our finding in cultured hippocampal neurons treated with oxygen/glucose deprivation (OGD) in vitro. Importantly, ENO1 overexpression in cultured neurons alleviated dendritic and spinal loss caused by OGD treatment. Furthermore, the enzymatic product of ENO1, phosphoenolpyruvate (PEP), was also synchronously changed along with the dynamic ENO1 level. The neuronal injury caused by OGD treatment in vitro or ischemia in vivo was mitigated by the application of PEP. Taken together, our data revealed that ENO1 plays a novel and protective role in cerebral ischemia-induced neuronal injury, highlighting a potential of ENO1 as a therapeutic target of neuronal protection from cerebral ischemia.
中风是全球成年人残疾和死亡的第二大主要原因，而神经元死亡和功能障碍的机制仍知之甚少。在这里，我们使用考马斯亮蓝染色，随后进行质谱分析，研究了 3 小时大脑中动脉闭塞 (MCAO) 缺血或假手术小鼠脑匀浆的差异蛋白质组图谱。我们发现烯醇酶 1 (ENO1)，一种关键的糖酵解酶，是 MCAO 缺血模型中神经元损伤的潜在介质。逆转录聚合酶链反应和蛋白质印迹数据显示，ENO1 在各种组织、大脑不同区域和不同出生后年龄中均有表达。免疫组织化学分析显示 ENO1 定位于神经元细胞质和树突中。有趣的是，ENO1 的表达水平在体内脑缺血的早期阶段显著增加，但在晚期阶段急剧下降。这种动态变化与我们在体外培养海马神经元中观察到的氧/葡萄糖剥夺 (OGD) 处理结果一致。重要的是，在体外培养神经元中过表达 ENO1 可减轻 OGD 处理引起的树突和脊髓丢失。此外，ENO1 的酶产物磷酸烯醇丙酮酸 (PEP) 也随着 ENO1 水平的动态变化而同步变化。体外 OGD 处理或体内缺血引起的神经元损伤可通过应用 PEP 得到缓解。总之，我们的数据表明 ENO1 在脑缺血引起的神经元损伤中发挥新的保护作用，突出了 ENO1 作为脑缺血神经元保护治疗靶点的潜力。
