Department of Ophthalmology, Tohoku University Graduate School of Medicine, 1-1 Seiryo-machi, Aoba-ku, Sendai, 980-8574, Japan.
Department of Retinal Disease Control, Tohoku University Graduate School of Medicine, Sendai, 980-8574, Japan.
Sci Rep. 2021 May 27;11(1):11159. doi: 10.1038/s41598-021-90604-w.
Administration of the mitochondrial complex I inhibitor rotenone provides an excellent model to study the pathomechanism of oxidative stress-related neural degeneration diseases. In this study, we examined the glial roles in retinal cell survival and degeneration under the rotenone-induced oxidative stress condition. Mouse-derived Müller, microglial (BV-2), and dissociated retinal cells were used for in vitro experiments. Gene expression levels and cell viability were determined using quantitative reverse transcription-polymerase chain reaction and the alamarBlue assay, respectively. Conditioned media were prepared by stimulating glial cells with rotenone. Retinal ganglion cells (RGCs) and inner nuclear layer (INL) were visualized on rat retinal sections by immunohistochemistry and eosin/hematoxylin, respectively. Rotenone dose-dependently induced glial cell death. Treatment with rotenone or rotenone-stimulated glial cell-conditioned media altered gene expression of growth factors and inflammatory cytokines in glial cells. The viability of dissociated retinal cells significantly increased upon culturing in media conditioned with rotenone-stimulated or Müller cell-conditioned media-stimulated BV-2 cells. Furthermore, intravitreal neurotrophin-5 administration prevented the rotenone-induced reduction of RGC number and INL thickness in rats. Thus, glial cells exerted both positive and negative effects on retinal cell survival in rotenone-induced neural degeneration via altered expression of growth factors, especially upregulation of microglia-derived Ntf5, and proinflammatory cytokines.
使用线粒体复合物 I 抑制剂鱼藤酮进行给药,为研究氧化应激相关神经退行性疾病的发病机制提供了一个极佳的模型。在这项研究中,我们研究了在鱼藤酮诱导的氧化应激条件下胶质细胞在视网膜细胞存活和变性中的作用。使用源自小鼠的 Muller 细胞、小胶质细胞(BV-2)和分离的视网膜细胞进行体外实验。使用定量逆转录聚合酶链反应和 alamarBlue 测定法分别确定基因表达水平和细胞活力。通过用鱼藤酮刺激神经胶质细胞来制备条件培养基。通过免疫组织化学和曙红/苏木精分别在大鼠视网膜切片上可视化视网膜神经节细胞(RGC)和内核层(INL)。鱼藤酮呈剂量依赖性诱导神经胶质细胞死亡。用鱼藤酮或鱼藤酮刺激的神经胶质细胞条件培养基处理会改变神经胶质细胞中生长因子和炎症细胞因子的基因表达。在用鱼藤酮刺激或 Muller 细胞条件培养基刺激的 BV-2 细胞的条件培养基中培养时,分离的视网膜细胞的活力显著增加。此外,玻璃体内给予神经营养因子-5 可防止鱼藤酮诱导的大鼠 RGC 数量和 INL 厚度减少。因此,神经胶质细胞通过改变生长因子的表达,特别是上调小胶质细胞衍生的 Ntf5 和促炎细胞因子,对鱼藤酮诱导的神经退行性变中的视网膜细胞存活产生了正反两方面的影响。