Functional analysis of mesencephalic astrocyte-derived neurotrophic factor in retinal ganglion cells under oxidative stress
Functional analysis of mesencephalic astrocyte-derived neurotrophic factor in retinal ganglion cells under oxidative stress
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DOI：
10.1002/cbf.3567
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发表时间：
2020-07-01
影响因子：
3.6
通讯作者：
Kiuchi, Yoshiaki
中科院分区：
文献类型：
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作者：
Ko, Ji-Ae;Komatsu, Kaori;Kiuchi, Yoshiaki
青光眼是以视网膜神经节细胞（RGC）和轴突的进行性神经变性为特征的视神经病变。这种情况将导致视力障碍，使青光眼成为全球第二大致盲原因。需要对青光眼进行神经保护以防止视神经病变的进展。在这项研究中，我们研究了上级丘（SC），中脑星形胶质细胞源性神经营养因子（MANF）分泌的SC，对RGC的生存氧化应激后的影响。使用3D-transwell系统共培养来自大鼠（3日龄）的SC切片和RGCs。此外，原代RGCs从4至5日龄的大鼠进行培养和处理与100 μ M过氧化氢（H2 O2），与刺激的MANF。免疫印迹和免疫荧光分析表明，下调的表达水平的几个生存标志物的RGCs。用100 μ M H2 O2处理的RGC中神经突的延伸减少。在与SC切片共培养或添加MANF后，我们发现RGCs中由氧化应激引起的神经标记物表达下调和神经突起延伸均被阻断。此外，我们发现与经MANF siRNA处理的SC切片共培养后，与经模拟siRNA处理的切片相比，神经标记物的表达和神经突起的延伸减少，但与经MANF siRNA处理的SC切片共培养的RGCs对凋亡无影响。这些结果表明，从SC分泌的MANF可能在维持RGCs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。研究意义青光眼是视网膜神经节细胞（RGC）及其轴突的进行性神经退行性疾病。神经保护的目的是保护那些受损的视神经元。我们现在已经研究了上级丘，或中脑星形胶质细胞源性神经营养因子（MANF），分泌从上级丘，对RGC生存的影响，使用共培养系统。提示MANF可能是RGC神经保护作用的重要因素。
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 mu M 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 mu M 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. Significance of the Study Glaucoma is a progressive neurodegenerative disease of the retinal ganglion cell (RGC) and their axons. Neuroprotection is aimed at protecting those neurons that are damaged glaucomatous optic neuropathy. We have now examined the effects of superior colliculus, or msencephalic astrocyte-derived neutrophic factor (MANF), secreted from superior colliculus, on RGC survival using co-culture system. Our results suggested that MANF may important key factor in neuroprotection of RGC.