[1]刘　明　杭春华　蔡智基　刘美秋　李　松　 吴光辉.Necrostatin-1在小鼠脊髓损伤后继发性损伤中的作用[J].中国临床神经外科杂志,2018,(06):419-422.[doi:10.13798/j.issn.1009-153X.2018.06.012]
　LIU Ming,HANG Chun-hua,CAI Zhi-ji,et al.Role of Necrostatin-1 in secondary injury after spinal cord injury in adult mice[J].,2018,(06):419-422.[doi:10.13798/j.issn.1009-153X.2018.06.012]
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Necrostatin-1在小鼠脊髓损伤后继发性损伤中的作用()
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《中国临床神经外科杂志》[ISSN:1009-153X/CN:42-1603/TN]

卷:
期数:
2018年06期
页码:
419-422
栏目:
论著
出版日期:
2018-06-25

文章信息/Info

Title:
Role of Necrostatin-1 in secondary injury after spinal cord injury in adult mice
文章编号:
1009-153X（2018）06-0419-04
作者:
刘　明　杭春华　蔡智基　刘美秋　李　松　 吴光辉
作者单位：352100 福建宁德， 福建医科大学附属宁德市医院神经外科（刘　明、蔡智基、刘美秋、吴光辉 ）；210002，南京，中国人民解放军南京总医院神经外科 （杭春华、李　松）
Author(s):
LIU Ming1;  HANG Chun-hua2;  CAI Zhi-ji1;  LIU Mei-qiu1;  LI Song2;  WU Guang-hui1.
1. Department of Neurosurgery, Ningde Municipal Hospital, Ningde 352100, China; 2. Department of Neurosurgery, Nanjing General Hospital, PLA, Nanjing 210002, China
关键词:
脊髓损伤; 受体相互作用蛋白;  活性氧簇; 线粒体; Necrostatin-1; 成年小鼠
Keywords:
Spinal cord injury;  Necrostatin-1;  Receptor interaction protein;  Secondary injury;  Mice
分类号:
R 641
DOI:
10.13798/j.issn.1009-153X.2018.06.012
文献标志码:
A
摘要:
目的 探讨Necrostatin-1(Nec-1)在小鼠脊髓损伤后继发性损伤中的作用及机制。方法 将168只健康成年雌性LCR小鼠随机分为4组：对照组（48只）、脊髓损伤组（48只）、溶剂组（36只，鞘内注射4 μl二甲基亚砜）和治疗组[36只，鞘内注射4 μl Nec-1（4 mmol/L）]。采用血管夹钳夹小鼠脊髓建立脊髓损伤模型。伤后6、12、24、48 h采用免疫印迹法检测脊髓组织受体相互作用蛋白（RIP）1、3的表达；伤后24 h采用免疫共沉淀法评估RIP1和RIP3的相互作用；伤后24 h检测脊髓组织丙二醛（MDA）和活性氧簇（ROS）水平，电镜观察小鼠脊髓组织神经元线粒体损伤情况。结果 伤后48 h内，小鼠脊髓RIP1表达水平无明显变化；伤后6 h，小鼠脊髓RIP3表达水平明显增高，持续到伤后48 h。伤后24 h，治疗组和溶剂组RIP1和RIP3的表达水平均无明显差异；正常脊髓组织RIP1和RIP3相互作用较弱，脊髓损伤后RIP1和RIP3相互作用加强，而Nec-1显著抑制RIP1和RIP3相互作用。伤后24 h，脊髓神经元线粒体不同程度受损，而治疗组小鼠脊髓神经元线粒体结构保存相对较好。伤后24 h，脊髓组织MDA和ROS含量明显升高，而Nec-1能明显减少小鼠脊髓MDA和ROS含量。结论 小鼠脊髓损伤后，Nec-1通过抑制RIP1和RIP3的相互作用，进而抑制程序性坏死，减轻脊髓继发性损伤。Nec-1能降低ROS产物，减轻氧化应激损伤，保护线粒体功能。
Abstract:
Objective To study the role of Necrostatin-1 (Nec-1) in secondary injury after spinal cord injury (SCI) in adult mice. Methods One hundred and sixty-eight female adult ICR mice were randomized divided into four groups, i.e. control group (n=48) and SCI group (n=48), vehicle group (n=36, intrathecal injection of 4 μl dimethyl sulfoxide) and treatment group (n=36, intrathecal injection of 4 μl Nec-1 with a concentration of 4 mmol/L). The SCI model was established by clamping the spinal cord with a vascular clamp. The expressions of receptor interaction protein (RIP)1 and RIP3 in the spinal cord tissues were determined by western blot 6, 12, 24 and 48 hours after SCI. The interaction of RIP1 and RIP3 was determined by immunoprecipitation 24 hours after SCI. The levels of malondiadehyde (MDA) and reactive oxygen species (ROS) in the spinal cord tissues was assessed. The ultrastructure of neuronal mitochondria in the spinal cord was observed by electron microscope 24 hours after SCI. Results Within 48 hours after SCI, there was no significant change in the expression of RIP1. Six hours after SCI, the expression of RIP3 increased significantly and continued until 48 hours after SCI. At 24 hours after SCI, the expression levels of RIP1 and RIP3 were not significantly different between the treatment and solvent groups. The interaction between RIP1 and RIP3 in the normal spinal cord tissue was weak, and was enhanced after SCI. Nec-1 significantly inhibited the interaction between RIP1 and RIP3. Twenty-four hours after SCI, the mitochondria of spinal cord neurons were damaged to certain degree, while the mitochondrial structure of spinal cord neurons in the treated group was preserved relatively well. At 24 h after SCI, the levels of MDA and ROS in spinal cord tissues increased significantly, while Nec-1 significantly decreased the levels of MDA and ROS. Conclusions Nec-1 reduces secondary injury by inhibiting the interaction between RIP1 and RIP3 and programmed necrosis after SCI in mice. Nec-1 can reduce ROS production, reduce oxidative stress and protect mitochondrial function.

参考文献/References:

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备注/Memo

备注/Memo:
基金项目：福建省卫生计生委青年科研课题
更新日期/Last Update: 2018-06-25