Delayed Administration of Bone Marrow Mesenchymal Stem Cell Conditioned Medium Significantly Improves Outcome After Retinal Ischemia in Rats
Delayed Administration of Bone Marrow Mesenchymal Stem Cell Conditioned Medium Significantly Improves Outcome After Retinal Ischemia in Rats
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DOI：
10.1167/iovs.13-11683
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发表时间：
2014-06-01
影响因子：
4.4
通讯作者：
Roth, Steven
中科院分区：
文献类型：
--
作者：
Dreixler, John C.;Poston, Jacqueline N.;Roth, Steven
目的。脑缺血后的延迟治疗往往不能令人满意。我们假设缺血后注射骨髓干细胞(BMSC)条件培养液可以挽救缺血的视网膜，在本研究中，我们研究了这种神经保护的功能和组织学结果及其机制。方法：成年Wistar大鼠通过升高眼压55分钟造成视网膜缺血。分别于缺血结束后24小时将大鼠骨髓间充质干细胞条件培养液(CM)或非条件培养液(UCM)注入玻璃体内。缺血7天后用视网膜电描记术评估视网膜的恢复情况，然后安乐死动物，制作4亩厚的石蜡包埋视网膜切片。分别于注射后24小时，即缺血后48小时，用原位末端标记法和Western印迹法检测细胞凋亡和凋亡相关基因的表达。结果缺血后2 4h玻璃体腔注射CM可显著改善视网膜功能，减轻视网膜神经节细胞层细胞丢失。减少脑缺血后细胞凋亡及凋亡相关基因的表达。通过光谱计数，与UCM相比，CM中符合严格鉴定标准的19个蛋白质增加；大多数是细胞外基质蛋白质，它们与其他参与细胞生长和黏附的蛋白质一起映射到一个相互作用的网络中。结论CM通过恢复视网膜功能，减弱细胞凋亡，防止缺血后视网膜细胞丢失，是一种强有力的延迟缺血后干预手段。我们确定了一些潜在的候选蛋白质来实现这种效应。
PURPOSE. Delayed treatment after ischemia is often unsatisfactory. We hypothesized that injection of bone marrow stem cell (BMSC) conditioned medium after ischemia could rescue ischemic retina, and in this study we characterized the functional and histological outcomes and mechanisms of this neuroprotection.METHODS. Retinal ischemia was produced in adult Wistar rats by increasing intraocular pressure for 55 minutes. Conditioned medium (CM) from rat BMSCs or unconditioned medium (uCM) was injected into the vitreous 24 hours after the end of ischemia. Recovery was assessed 7 days after ischemia using electroretinography, at which time we euthanized the animals and then prepared 4-mu m-thick paraffin-embedded retinal sections. TUNEL and Western blot were used to identify apoptotic cells and apoptosis-related gene expression 24 hours after injections; that is, 48 hours after ischemia. Protein content in CM versus uCM was studied using tandem mass spectrometry, and bioinformatics methods were used to model protein interactions.RESULTS. Intravitreal injection of CM 24 hours after ischemia significantly improved retinal function and attenuated cell loss in the retinal ganglion cell layer. CM attenuated postischemic apoptosis and apoptosis-related gene expression. By spectral counting, 19 proteins that met stringent identification criteria were increased in the CM compared to uCM; the majority were extracellular matrix proteins that mapped into an interactional network together with other proteins involved in cell growth and adhesion.CONCLUSIONS. By restoring retinal function, attenuating apoptosis, and preventing retinal cell loss after ischemia, CM is a robust means of delayed postischemic intervention. We identified some potential candidate proteins for this effect.