Neuroserpin restores autophagy and promotes functional recovery after acute spinal cord injury in rats.
Neuroserpin restores autophagy and promotes functional recovery after acute spinal cord injury in rats.
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Neuroserpin恢复大鼠急性脊髓损伤后自噬并促进功能恢复
DOI：
10.3892/mmr.2017.8249
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发表时间：
2018-03
影响因子：
3.4
通讯作者：
Chen Z
中科院分区：
文献类型：
--
作者：
Li Z;Liu F;Zhang L;Cao Y;Shao Y;Wang X;Jiang X;Chen Z
本研究旨在揭示脊髓损伤后功能恢复过程中自噬的特点及神经丝氨酸蛋白酶抑制剂（NSP）对自噬的影响。制作大鼠脊髓夹压模型后，分别于2、4、24、72、168 h检测实验组和假手术组大鼠脊髓自噬相关蛋白LC 3-II、beclin-1和p62的表达。透射电子显微镜（TEM）进一步用于在4和72 h的自噬检测。所有雄性大鼠随机分为3组：假手术组、溶剂组和NSP组。SCI后立即通过鞘内注射给予NSP或等体积的生理盐水载体。每组进一步分成亚组用于以下实验：i）蛋白质印迹（LC 3-II和p62）; ii）免疫荧光双重染色（LC 3/MAP-2/DAPI）; iii）Nissl染色和Basso Beattie Bresnahan（BBB评分）用于NSP神经保护评价。我们的研究结果显示，LC 3-II和p62的表达在SCI后24，72和168小时呈上升趋势。LC 3-II在72 h达到峰值，而p62在24 h达到峰值。Beclin-1在72和168 h显著下降。透射电镜结果显示，与假手术组相比，脊髓损伤后72 h自噬体大量聚集。Western blot分析显示，与赋形剂组相比，在损伤后72 h，NSP处理的LC 3-II和p62显著降低。免疫荧光双标记表明，自噬体的积累减少在NSP组。此外，SCI后用NSP治疗改善了BBB评分，并增加了前角运动神经元的数量。总之，这项研究表明，自噬流量受损，同时NSP恢复自噬流量，促进SCI后大鼠的功能恢复。
This study is to reveal the characteristics of autophagy and the effect of neuroserpin (NSP) treatment on autophagy during the process of functional recovery following spinal cord injury (SCI). After the clip compress rat model of SCI had been made, autophagy-associated proteins, including LC3-II, beclin-1 and p62, were evaluated at 2, 4, 24, 72 h, and 168 h in the experimental group, and the sham group as control. Transmission electron microscopy (TEM) was further used for autophagy detection at 4 and 72 h. All the male rats were randomly divided into three groups: Sham, vehicle and NSP group. NSP or an equal volume of saline vehicle was administered via intrathecal injection immediately after SCI. Each group was further divided into subgroups for the following experiments: i)Western blot (LC3-II and p62); ii) Immunofluorescent double staining (LC3/MAP-2/DAPI); iii) Nissl staining and Basso Beattie Bresnahan (BBB score) for NSP neuroprotection evaluation. Our results revealed both LC3-II and p62 expression trended upward at 24, 72 and 168 h after SCI. The LC3-II peaked at 72 h, while p62 peaked at 24 h. Beclin-1 dropped significantly at 72 and 168 h. TEM results showed that autophagosomes largely accumulated at 72 h after SCI when compared with the sham group. Western blot analysis showed that LC3-II and p62 were markedly decreased with NSP treatment at 72 h after injury compared with that of the vehicle-group. Immunofluorescent double labeling indicated that accumulation of autophagosomes was reduced in the NSP group. Further, post-SCI treatment with NSP improved the BBB scale and increased the number of anterior horn motor neurons. Together, this study demonstrates that autophagic flux is impaired, meanwhile NSP restores autophagic flux and promotes functional recovery after SCI in rats.
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影响因子：
6.1
作者：
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通讯作者：
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影响因子：
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作者：
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