神经干细胞与大鼠RPE细胞间的线粒体转运及对RPE细胞凋亡的影响

发布时间：2018-04-21 05:05

本文选题：干细胞 + 视网膜色素变性　；参考：《第三军医大学学报》2017年11期

【摘要】：目的研究小鼠神经干细胞(neural stem cells,NSCs)C17.2细胞系与从RCS大鼠变性视网膜中原代分离视网膜色素上皮细胞(retinal pigment epithelial,RPE)的线粒体交换方式及其对RPE细胞特性的影响。方法分离RCS大鼠RPE细胞并进行培养和鉴定。分别用线粒体特异性标记物Mitotracker-red和Mitotracker-green标记RPE细胞和小鼠NSCs细胞的线粒体,将两种细胞共培养,激光共聚焦显微镜下观察两种细胞之间隧道纳米管(tunneling nanotubes,TNT)的形成及线粒体转运方式。采用流式细胞仪检测与NSCs共培养后RPE细胞的反应性活性氧类(reactive oxygen species,ROS)水平、细胞周期和细胞凋亡水平的变化。结果来源于RCS大鼠视网膜的第3代RPE细胞生长状态良好,RPE65及Bestrophin蛋白阳性率细胞均大于95%。与NSCs共培养24 h后,可见RPE细胞与NSCs间TNT的形成,NSCs中的线粒体向RPE细胞方向单向运动,接受NSCs转运的线粒体后,RPE细胞的ROS水平降低(P0.01);RPE细胞的增殖能力增加,处于S期的细胞比例明显增加(P0.01),细胞增殖指数(PI)显著增加(P0.05);RPE细胞早期凋亡细胞比例显著降低(P0.05)。结论 NSCs可通过与相邻的变性RPE细胞之间形成TNT并向其转运线粒体而改善变性RPE细胞的存活。
[Abstract]:Objective to study the mitochondrial exchange patterns of neural stem cells from mouse neural stem cells and the primary isolation of retinal pigment epithelial cells (RPE) from the degenerative retina of RCS rats and their effects on the characteristics of RPE cells. Methods RCS rat RPE cells were isolated, cultured and identified. Mitochondrial specific markers (Mitotracker-red and Mitotracker-green) were used to label the mitochondria of RPE cells and mouse NSCs cells, respectively. The formation of tunneling nanotubes and mitochondrial transport patterns were observed under confocal laser microscope. The levels of reactive reactive oxygen species (Ros), cell cycle and apoptosis of RPE cells after co-culture with NSCs were detected by flow cytometry. Results the positive rates of RPE65 and Bestrophin protein in the third passage of RPE cells from the retina of RCS rats were higher than 95%. After co-cultured with NSCs for 24 h, the formation of TNT between RPE cells and NSCs cells showed that the mitochondria in RPE cells moved in one direction to RPE cells, and the ROS level of TNT cells after NSCs transport decreased the proliferation ability of P0.01RPE cells. The percentage of cells in S phase increased significantly, and the proliferation index (Pi) increased significantly. The percentage of early apoptosis cells in P0.05 RPE cells decreased significantly. Conclusion NSCs can improve the survival of denatured RPE cells by forming TNT between them and transporting mitochondria to them.
【作者单位】：第三军医大学西南医院全军眼科中心视觉损伤与再生修复重庆市重点实验室;
【基金】：国家重点基础研究发展计划(973计划,2013CB967002)~~
【分类号】：R774.1

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