Neurite regeneration in adult rat retinas exposed to advanced glycation end-products and regenerative effects of neurotrophin-4
Neurite regeneration in adult rat retinas exposed to advanced glycation end-products and regenerative effects of neurotrophin-4
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DOI：
10.1016/j.brainres.2013.08.027
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发表时间：
2013-10-09
期刊：
影响因子：
2.9
通讯作者：
Yamamoto, Shuichi
中科院分区：
文献类型：
--
作者：
Bikbova, Guzel;Oshitari, Toshiyuki;Yamamoto, Shuichi
本研究旨在探讨低浓度糖基化终末产物对大鼠视网膜轴突再生的影响，以及神经营养因子-4对晚期糖基化终末产物视网膜再生的影响。将4只成年SD大鼠的视网膜外植体接种于胶原胶上，分别在无血清对照、葡萄糖糖基化终产物-牛血清白蛋白、甘油醛糖基化终产物-牛血清白蛋白、甘油醛糖基化终产物-牛血清白蛋白、葡萄糖糖基化终产物+神经营养素-4、乙醇醛-糖基化终产物+神经营养素-4、甘油醛-糖基化终产物+神经营养素-4培养液中培养。7d后，计数外植体再生的轴突数量。然后，将外植体固定，冷冻，并进行TUNEL染色。测定神经节细胞层TUNEL阳性细胞占全部细胞的比例。免疫组织化学方法检测caspase-9活性形式和凋亡诱导因子的表达。在含有晚期糖基化终产物的视网膜培养液中，再生的轴突数量少于没有晚期糖基化终产物的视网膜，而TUNEL阳性细胞和caspase-9及凋亡诱导因子免疫阳性细胞的数量显著高于对照培养液。补充神经营养素-4可增加再生神经炎的数量，TUNEL阳性细胞数、caspase-9阳性细胞数和凋亡诱导因子免疫阳性细胞数显著少于未加神经营养素-4的晚期糖基化终末产物组。低剂量的高级糖基化终末产物阻碍了大鼠视网膜中的轴突再生。神经营养素-4显著促进接触晚期糖基化终末产物的视网膜中的轴突再生。(C)2013爱思唯尔B.V.保留所有权利。
The purpose of this study was to determine the effect of low concentrations of advanced glycation end-products on neurite regeneration in isolated rat retinas, and to determine the effects of neurotrophin-4 on regeneration in advanced glycation end-products exposed retinas. Retinal explants of 4 adult Sprague-Dawley rats were cultured on collagen gel and were incubated in; (1) serum-free control culture media, (2) glucose-advanced glycation end-products-bovine serum albumin media, (3) glycolaldehyde-advanced glycation end-products-bovine serum albumin media, (4) glyceraldehyde-advanced glycation end-products-bovine serum albumin media, (5) glucose-advanced glycation end-products+neurotrophin-4 media, (6) glycolaldehyde-advanced glycation end-products+neurotrophin-4 media, or (7) glyceraldehyde-advanced glycation end-products+neurotrophin-4 supplemented culture media. After 7 days, the number of regenerating neurites from the explants was counted. Then, explants were fixed, cryosectioned, and stained for TUNEL. The ratio of TUNEL-positive cells to all cells in the ganglion cell layer was determined. Immunohistochemical examinations for the active-form of caspase-9 and apoptosis-inducing factor were performed. In retinas incubated with advanced glycation end-products containing media, the number of regenerating neurites were fewer than in retinas without advanced glycation end-products, and the number of TUNEL-positive cells and caspase-9- and apoptosis-inducing factor-immunopositive cells was significantly higher than in control media. Neurotrophin-4 supplementation increased the numbers of regenerating neuritis, and the number of TUNEL-positives, caspase-9-, and apoptosis-inducing factor-immunopositive cells were significantly fewer than that in advanced glycation end-products without neurotrophin-4 media. Low doses of advanced glycation end-products impede neurite regeneration in the rat retinas. Neurotrophin-4 significantly enhances neurite regeneration in retinas exposed to advanced glycation end-products. (C) 2013 Elsevier B.V. All rights reserved.