中国组织工程研究

中国组织工程研究 ›› 2021, Vol. 25 ›› Issue (26): 4175-4179.doi: 10.12307/2021.117

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

脂氧素A4对脊髓缺血再灌注损伤模型大鼠的神经保护

路  坦1，常耀辉2，尉  娜3   

  1. 1新乡医学院第一附属医院骨外二科，河南省卫辉市   453100；2修武县人民医院骨外科，河南省焦作市   454350；3新乡医学院第三附属医院神经内二科，河南省新乡市   453003
  • 收稿日期:2019-04-01 修回日期:2019-05-06 接受日期:2020-08-11 出版日期:2021-09-18 发布日期:2021-05-10
  • 通讯作者: 尉娜，副主任医师，新乡医学院第三附属医院神经内二科，河南省新乡市 453003
  • 作者简介:路坦，男，1982年生，河南省林州市人，汉族，博士，副主任医师，硕士生导师，主要从事脊髓损伤修复方面的研究。
  • 基金资助:
    河南省高等学校重点科研项目(19A320003)，项目负责人：路坦；新乡市科技攻关计划项目(CXGG17031)，项目负责人：尉娜；新乡医学院第一附属医院青年基金项目(QN-2017-A002)，项目负责人：路坦

Lipoxanthin A4 plays a neuroprotective role against spinal cord ischemia-reperfusion injury in rats

Lu Tan1, Chang Yaohui2, Wei Na3   

  1. 1Department of Orthopedics, the First Affiliated Hospital of Xinxiang Medical University, Weihui 453100, Henan Province, China; 2Department of Orthopedics, Xiuwu County People’s Hospital, Jiaozuo 454350, Henan Province, China; 3Department of Neurology, the Third Affiliated Hospital of Xinxiang Medical University, Xinxiang 453003, Henan Province, China
  • Received:2019-04-01 Revised:2019-05-06 Accepted:2020-08-11 Online:2021-09-18 Published:2021-05-10
  • Contact: Wei Na, Associate chief physician, Department of Neurology, the Third Affiliated Hospital of Xinxiang Medical University, Xinxiang 453003, Henan Province, China
  • About author:Lu Tan, MD, Associate chief physician, Master’s supervisor, Department of Orthopedics, the First Affiliated Hospital of Xinxiang Medical University, Weihui 453100, Henan Province, China
  • Supported by:
    the Key Scientific Research Project of Henan Provincial Colleges and Universities, No. 19A320003 (to LT); Xinxiang Municipal Science and Technology Research Project, No. CXGG17031 (to WN); Youth Fund of the First Affiliated Hospital of Xinxiang Medical University, No. QN-2017-A002 (to LT)

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摘要：

文题释义：
脊髓缺血再灌注损伤：在脊髓损伤长期受压后，当脊髓受压解除血液再通后继而出现神经损伤症状，其本质是当脊髓血液再通后多种因素造成不可逆性神经元死亡，包括二次打击。
脂氧素：是一类由花生四烯酸代谢而来的内源性脂类介质，主要作用是促进炎症反应的及时消退，被称为炎症反应的“刹车信号”，可以明显抑制白细胞的驱化和激活，是体内重要的抗炎递质。

背景：前期研究已证明脂氧素A4对大鼠脊髓损伤具有神经保护作用，但其在脊髓缺血再灌注损伤中的作用机制仍不明确。
目的：探讨脂氧素A4在大鼠脊髓缺血再灌注损伤中的保护作用。
方法：将48只SD大鼠随机分为3组，每组16只：对照组、实验组制作脊髓缺血再灌注损伤模型，实验组脊髓鞘内注射脂氧素A4，对照组脊髓鞘内注射生理盐水；空白组不造模。造模后6，12，24，48 h，利用BBB评分评估大鼠运动神经功能；再灌注后24 h，采用Tunel法观察脊髓组织细胞凋亡情况，ELISA法检测脊髓组织Bcl-2和Bax浓度，Western blot检测脊髓组织cleaved-caspase-3、p-IKKβ和p-NF-κB蛋白的表达。实验通过新乡医学院第一附属医院伦理委员会批准(2018038)。
结果与结论：①对照组不同时间点的BBB评分低于空白组(P < 0.05)，实验组造模后12，24，48 h的BBB评分高于对照组(P < 0.05)；②对照组细胞凋亡多于空白组(P < 0.05)，实验组少于对照组(P < 0.05)；③对照组脊髓组织内的Bcl-2、Bax水平均高于空白组(P < 0.05)；实验组的Bcl-2水平高于对照组(P < 0.05)，Bax水平低于对照组(P < 0.05)；④对照组cleaved-caspase-3、p-NF-κB蛋白表达强于空白组，p-IKKβ蛋白表达弱于空白组；实验组cleaved-caspase-3、p-NF-κB蛋白表达弱于对照组，p-IKKβ蛋白表达强于对照组；⑤结果表明，脂氧素A4可激活细胞质中caspase-3/IKKβ/NF-κB通道的活化，抑制细胞凋亡，起到治疗脊髓缺血再灌注损伤的作用。
https://orcid.org/0000-0001-5493-814X (路坦)

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 组织构建, 脊髓, 脂氧素A4, 脊髓缺血再灌注损伤, 细胞凋亡, 信号通路

Abstract:

BACKGROUND: Previous studies have indicated that lipoxin A4 has a neuroprotective effect against spinal cord injury in rats, but its mechanism of action in spinal cord ischemia-reperfusion injury is still unclear.

OBJECTIVE: To investigate the neuroprotective effect of lipoxin A4 in rats with spinal cord ischemia-reperfusion injury. 
METHODS: Forty-eight Sprague-Dawley rats were randomized into three groups (n=16 per group): Spinal cord ischemia-reperfusion injury models were built in control and experimental groups followed by intrathecal injection of normal saline and lipoxin A4 respectively, and no treatment was given in blank control group. Basso-Beattie-Bresnahan (BBB) score was used to evaluate motor nerve function in rats at 6, 12, 24, and 48 hours after modeling. At 24 hours after reperfusion, TUNEL assay was used to detect cell apoptosis in the rat spinal cord; ELISA was used to detect the levels of Bcl-2 and Bax in the rat spinal cord, and western blot assay was used to detect the levels of cleaved-caspase-3, p-IKKβ and p-NF-κB proteins in the rat spinal cord. This experiment was approved by the Ethics Committee of the First Affiliated Hospital of Xinxiang Medical University (approval No. 2018038).
RESULTS AND CONCLUSION: Compared with the blank control group, the BBB scores were lower in the control group at different observation time points 
(P < 0.05), while compared with the control group, the BBB score was higher in the experimental group at 12, 24, and 48 hours after modeling (P < 0.05). There were more apoptotic cells in the control group than the blank control group (P < 0.05), while the number of apoptotic cells in the experimental group was lower than that in the control group (P < 0.05). The levels of Bcl-2 and Bax in the spinal cord were significantly higher in the control group than the blank control group, while compared with the control group, the experimental group showed a higher Bcl-2 level and a lower Bax level (P < 0.05). Compared with the blank control group, the expressions of cleaved-caspase-3 and p-NF-κB proteins were strongly higher in the control group, while the protein expression of p-IKKβ was weaker in the control group. Compared with the control group, the expressions of cleaved-caspase-3 and p-NF-κB proteins were lower in the experimental group, while the protein expression of p-IKKβ was higher in the experimental group. These findings indicate that lipoxin A4, as a therapeutic agent for spinal cord ischemia-reperfusion injury, can inhibit cell apoptosis by activating the Caspase-3/IKKβ/NF-κB pathway in the cytoplasm.

Key words: tissue construction, spinal cord, lipoxin A4, spinal cord ischemia-reperfusion injury, apoptosis, signaling pathway

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