The School of Life Sciences, Henan University, 475000, Henan Province, Kaifeng, P. R. China.
CNS Neurosci Ther. 2023 Sep;29(9):2583-2596. doi: 10.1111/cns.14198. Epub 2023 Apr 5.
To explore whether 14, 15-EET regulates mitochondrial dynamics to exert neuroprotective effects after cerebral ischemia-reperfusion and its underlying mechanisms.
The mouse middle cerebral artery occlusion reperfusion model was used to observe brain infarct volume and neuronal apoptosis by TTC staining and Tunel assay, modified neurological severity score to detect neurological impairment, HE staining and Nissl staining to observe neuron damage, western blot and immunofluorescence methods to detect the expression of mitochondrial dynamics-related proteins, transmission electron microscopy, and Golgi-Cox staining to detect mitochondrial morphology and neuronal dendritic spines.
14, 15-EET reduced the neuronal apoptosis and cerebral infarction volume induced by middle cerebral artery occlusion reperfusion (MCAO/R), inhibited the degradation of dendritic spines, maintained the structural integrity of neurons, and alleviated neurological impairment. Cerebral ischemia-reperfusion induces mitochondrial dynamics disorders, upregulates the expression of the mitochondrial division protein Fis 1, and inhibits the expression of mitochondrial fusion proteins MFN1, MFN2, and OPA1, while 14, 15-EET treatment reverses this process. Mechanistic studies have shown that 14, 15-EET promotes the phosphorylation of AMPK, upregulates the expression of SIRT1 and phosphorylation of FoxO1, thereby inhibiting mitochondrial division and promoting mitochondrial fusion, preserving mitochondrial dynamics, maintaining neuronal morphological and structural integrity, and alleviating neurological impairment induced by middle cerebral artery occlusion reperfusion. Compound C treatment diminishes the neuroprotective effect of 14, 15-EET following MCAO/R in mice.
This study elucidates the novel neuroprotective mechanism of 14, 15-EET, providing a novel approach for the development of drugs based on mitochondrial dynamics.
探讨 14,15-EET 是否通过调节线粒体动力学发挥脑缺血再灌注后的神经保护作用及其机制。
采用小鼠大脑中动脉闭塞再灌注模型，通过 TTC 染色和 Tunel 检测观察脑梗死体积和神经元凋亡，改良神经功能缺损评分检测神经损伤，HE 染色和尼氏染色观察神经元损伤，Western blot 和免疫荧光法检测线粒体动力学相关蛋白的表达，透射电镜和 Golgi-Cox 染色检测线粒体形态和神经元树突棘。
14,15-EET 减少大脑中动脉闭塞再灌注（MCAO/R）引起的神经元凋亡和脑梗死体积，抑制树突棘降解，维持神经元结构完整性，减轻神经损伤。脑缺血再灌注诱导线粒体动力学紊乱，上调线粒体分裂蛋白 Fis1 的表达，抑制线粒体融合蛋白 MFN1、MFN2 和 OPA1 的表达，而 14,15-EET 处理可逆转这一过程。机制研究表明，14,15-EET 促进 AMPK 磷酸化，上调 SIRT1 和 FoxO1 磷酸化表达，抑制线粒体分裂，促进线粒体融合，维持线粒体动力学，保持神经元形态和结构的完整性，减轻 MCAO/R 引起的神经损伤。复合物 C 处理可减弱 14,15-EET 对 MCAO/R 后小鼠的神经保护作用。
本研究阐明了 14,15-EET 的新型神经保护机制，为基于线粒体动力学的药物开发提供了新途径。
