Glaucoma is optic neuropathy that is characterized by progressive neurodegeneration of the retinal ganglion cells (RGCs) and axons. This condition will lead to visual impairment and bring glaucoma to become the second cause of blindness globally. Neuroprotection in glaucoma is needed to prevent the progression of optic neuropathy. In this study, we examined the effects of the superior colliculus (SC), and mesencephalic astrocyte‐derived neurotrophic factor (MANF) secreted from the SC, on RGC survival after oxidative stress. SC slices and RGCs from rats (3‐day old) were co‐cultured using a 3D‐transwell system. In addition, primary RGCs from 4 to 5‐day‐old rats were cultured and treated with 100 μM hydrogen peroxide (H₂O₂), together with stimulation by MANF. Immunoblot and immunofluorescence analyses indicated down‐regulated expression levels of several survival markers of RGCs. Extension of neurites was decreased in RGCs treated with 100 μM H₂O₂. Following co‐culture with SC slices, or the addition of MANF, we found that both the down‐regulated expression of neural markers and extension of neurites caused by oxidative stress in RGCs were blocked. Furthermore, we found a decrease in the expression of neural markers and extension of neurites after co‐culture with MANF siRNA‐treated SC slices compared with slices treated with mock siRNA, but, RGCs co‐cultured with SC slices treated with MANF siRNA displayed no‐changed about to apoptosis. These results suggest that MANF secreted from the SC may play an important role in maintenance of function and survival of RGCs. It is also possible that MANF is an important factor in neuroprotection of RGCs.

中文翻译：

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氧化应激下视网膜神经节细胞中脑星形胶质细胞源性神经营养因子的功能分析。

青光眼是一种视神经病变，其特征在于视网膜神经节细胞（RGC）和轴突的进行性神经变性。这种情况将导致视力障碍，并使青光眼成为全球失明的第二大原因。需要青光眼的神经保护以防止视神经病变的发展。在这项研究中，我们研究了上丘（SC）和SC分泌的中脑星形胶质细胞源性神经营养因子（MANF）对氧化应激后RGC存活的影响。使用3D Transwell系统将大鼠（3天大）的SC切片和RGC进行共培养。此外，对4至5天大的大鼠原代RGC进行培养，并用100μM过氧化氢（H ₂ O ₂），以及MANF的刺激。免疫印迹和免疫荧光分析表明，RGC的几种存活标志物的表达水平下调。用100μMH ₂ O ₂处理的RGC中神经突的延伸减少。与SC切片共培养或添加MANF后，我们发现RGC中神经标记的下调表达和氧化应激引起的神经突延伸均被阻止。此外，与用模拟siRNA处理的切片相比，与MANF siRNA处理的SC切片共培养后，我们发现神经标志物的表达和神经突的延伸减少，但是，与经MANF siRNA处理的SC切片共培养的RGC没有显示出即将发生凋亡。这些结果表明，SC分泌的MANF可能在RGC的功能和存活中起重要作用。MANF也可能是RGC神经保护的重要因素。