Intravitreal administration of multipotent mesenchymal stromal cells triggers a cytoprotective microenvironment in the retina of diabetic mice.
Intravitreal administration of multipotent mesenchymal stromal cells triggers a cytoprotective microenvironment in the retina of diabetic mice.
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DOI：
10.1186/s13287-016-0299-y
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发表时间：
2016-03-16
影响因子：
7.5
通讯作者：
Ezquer F
中科院分区：
文献类型：
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作者：
Ezquer M;Urzua CA;Montecino S;Leal K;Conget P;Ezquer F
糖尿病视网膜病变是糖尿病的常见并发症，也是西方世界不可逆视力丧失的主要原因。由于视网膜的神经节细胞层中的神经细胞的损失而导致的颜色/对比敏感度的降低是糖尿病性视网膜病变发作的早期事件。多能间充质基质细胞（Multipotent mesenchymal stromal cells，MSCs）具有向神经元细胞分化、产生高水平神经营养因子和降低氧化应激等特点，是治疗神经退行性疾病的理想工具。我们的目的是确定玻璃体内注射脂肪来源的MSC是否能够预防糖尿病小鼠视网膜神经节细胞的丢失。通过施用链脲佐菌素在C57 BL 6小鼠中诱导糖尿病。当存在视网膜促损伤机制时，动物接受单次玻璃体内剂量的2 × 105脂肪来源的MSC或溶剂。4周和12周后，我们评估：（a）视网膜神经节细胞数量（免疫荧光）;（B）神经营养因子水平（实时定量聚合酶链反应（RT-qPCR）和酶联免疫吸附测定（ELISA））;（c）视网膜凋亡率（TUNEL）;（d）活性氧和氧化损伤的视网膜水平（ELISA）;（e）视网膜的电反应（f）促血管生成和抗血管生成因子水平（RT-qPCR和ELISA）;和（g）视网膜血管（血管造影术）。此外，MSC给药后1、4、8和12周，评估视网膜中供体细胞的存在及其向神经和血管周围样细胞的分化（免疫荧光和流式细胞术）。MSC施用完全防止视网膜神经节细胞损失。供体细胞留在玻璃体腔内，不分化为神经或血管周围样细胞。然而，它们增加了几种有效的神经营养因子（神经生长因子，碱性成纤维细胞生长因子和胶质细胞系衍生的神经营养因子）的眼内水平，并减少了视网膜中的氧化损伤。此外，MSC给药对视网膜的电反应具有中性作用，并且不会导致病理性新生血管形成。玻璃体内注射脂肪来源的间充质干细胞在糖尿病小鼠视网膜中触发有效的细胞保护微环境。因此，MSC代表了一种有趣的工具，以预防糖尿病视网膜病变。本文的在线版本（doi：10.1186/s13287-016-0299-y）包含补充材料，可供授权用户使用。
Diabetic retinopathy is a common complication of diabetes and the leading cause of irreversible vision loss in the Western world. The reduction in color/contrast sensitivity due to the loss of neural cells in the ganglion cell layer of the retina is an early event in the onset of diabetic retinopathy. Multipotent mesenchymal stromal cells (MSCs) are an attractive tool for the treatment of neurodegenerative diseases, since they could differentiate into neuronal cells, produce high levels of neurotrophic factors and reduce oxidative stress. Our aim was to determine whether the intravitreal administration of adipose-derived MSCs was able to prevent the loss of retinal ganglion cells in diabetic mice. Diabetes was induced in C57BL6 mice by the administration of streptozotocin. When retinal pro-damage mechanisms were present, animals received a single intravitreal dose of 2 × 105 adipose-derived MSCs or the vehicle. Four and 12 weeks later we evaluated: (a) retinal ganglion cell number (immunofluorescence); (b) neurotrophic factor levels (real-time quantitative polymerase chain reaction (RT-qPCR) and enzyme-linked immunosorbent assay (ELISA)); (c) retinal apoptotic rate (TUNEL); (d) retinal levels of reactive oxygen species and oxidative damage (ELISA); (e) electrical response of the retina (electroretinography); (f) pro-angiogenic and anti-angiogenic factor levels (RT-qPCR and ELISA); and (g) retinal blood vessels (angiography). Furthermore, 1, 4, 8 and 12 weeks post-MSC administration, the presence of donor cells in the retina and their differentiation into neural and perivascular-like cells were assessed (immunofluorescence and flow cytometry). MSC administration completely prevented retinal ganglion cell loss. Donor cells remained in the vitreous cavity and did not differentiate into neural or perivascular-like cells. Nevertheless, they increased the intraocular levels of several potent neurotrophic factors (nerve growth factor, basic fibroblast growth factor and glial cell line-derived neurotrophic factor) and reduced the oxidative damage in the retina. Additionally, MSC administration has a neutral effect on the electrical response of the retina and did not result in a pathological neovascularization. Intravitreal administration of adipose-derived MSCs triggers an effective cytoprotective microenvironment in the retina of diabetic mice. Thus, MSCs represent an interesting tool in order to prevent diabetic retinopathy. The online version of this article (doi:10.1186/s13287-016-0299-y) contains supplementary material, which is available to authorized users.