The autophagy/apoptosis interaction has always been a focus of study in pathogenicity models. Neuritin is a neurotrophic factor that is highly expressed primarily in the central nervous system. Our previous study revealed that it protects against apoptosis in cortical neurons subjected to oxygen-glucose deprivation (OGD)/reoxygenation (OGD/R), and later animal experiments revealed that it can increase the expression of the autophagy-related protein LC3. Whether this neuroprotective effect is closely related to autophagy is still unclear. In this study, we hypothesized that neuritin can promote autophagic flux to protect nerve cells after OGD/R. To verify this hypothesis, we induced OGD/R in primary cortical neurons and assessed cell viability by the CCK8 and LDH assays. Cell apoptosis was assessed by Annexin V-FITC/PI, staining, and the contents and mRNA abundances of the autophagy-related proteins LC3 and p62, the apoptotic protein Caspase3 were quantified by Western blotting and RT-PCR. Autophagic flux was assessed by immunofluorescence after RFP-GFP-LC3 virus transfection, and ultrastructural changes in autophagosomes were observed by transmission electron microscopy (TEM). The results showed that cell viability was decreased, apoptosis was increased and autophagy was enhanced after OGD/R. Neuritin significantly increased cell viability, decreased apoptosis, further increased the expression of the autophagic flux-related protein LC3, further decreased p62 expression, and significantly increased the autophagosome number and autophagosome to lysosome ratio. Bafilomycin A1 (BafA1) is a late autophagy inhibitor, aggravated cell damage and apoptosis and counteracted the enhancement of autophagy activation and protective effects of neuritin. In conclusion, neuritin may promote the completion of autophagic flux by ameliorating neuronal damage after OGD/R.

自噬/凋亡相互作用一直是致病性模型研究的重点。神经蛋白是一种主要在中枢神经系统中高度表达的神经营养因子。我们之前的研究表明，它可以防止遭受氧葡萄糖剥夺（OGD）/复氧（OGD/R）的皮层神经元凋亡，后来的动物实验表明，它可以增加自噬相关蛋白 LC3 的表达。这种神经保护作用是否与自噬密切相关尚不清楚。在这项研究中，我们假设在 OGD/R 后，neuritin 可以促进自噬流以保护神经细胞。为了验证这一假设，我们在原代皮层神经元中诱导了 OGD/R，并通过 CCK8 和 LDH 测定评估了细胞活力。通过膜联蛋白 V-FITC/PI、染色、自噬相关蛋白LC3和p62、凋亡蛋白Caspase3的含量和mRNA丰度通过Western blotting和RT-PCR进行定量。RFP-GFP-LC3病毒转染后通过免疫荧光评估自噬通量，并通过透射电子显微镜（TEM）观察自噬体的超微结构变化。结果表明，OGD/R后细胞活力降低，细胞凋亡增加，自噬增强。Neuritin 显着增加细胞活力，减少细胞凋亡，进一步增加自噬通量相关蛋白 LC3 的表达，进一步降低 p62 的表达，并显着增加自噬体数量和自噬体与溶酶体的比例。Bafilomycin A1 (BafA1) 是一种晚期自噬抑制剂，加重细胞损伤和凋亡，并抵消了神经蛋白自噬激活和保护作用的增强。综上所述，neuritin 可能通过改善 OGD/R 后的神经元损伤来促进自噬通量的完成。