建立兔脊髓缺血再灌注损伤模型PDIA3的表达变化及临床意义

发布时间：2018-02-23 07:07

  本文关键词： 脊髓 缺血 再灌注损伤 蛋白质二硫键异构酶 组织构建 组织工程 脊髓缺血再灌注损伤 蛋白质二硫键异构酶A 应激 国家自然科学基金　出处：《中国组织工程研究》2017年24期 　论文类型：期刊论文

【摘要】：背景:目前认为脊髓缺血再灌注损伤是造成脊髓减压手术后"二次瘫痪"的主要原因,而控制应激相关蛋白和兴奋性氨基酸对脊髓缺血再灌注损伤的治疗至关重要。目的:观察蛋白质二硫键异构酶A3(PDIA3)在兔脊髓缺血再灌注损伤后表达变化。方法:依据Zivin法建立兔脊髓缺血再灌注损伤模型,将36只新西兰白兔随机均分成6组,对照组只显露腹主动脉而不阻断血流,30 min后关闭腹腔;实验组阻断腹主动脉血流30 min后分别再灌注0,6,12,24,48 h再关闭腹腔。首先均利用改良Tarlov评分进行运动功能评价;随后取损伤段腰髓(L3-L5),结合荧光差异双向凝胶电泳和质谱分析筛选出PDIA3;其次应用免疫印迹印证质谱;最后通过免疫组织化学结果观察其在脊髓内的时间和空间表达变化特点。结果与结论:(1)运动功能评分结果:实验动物于脊髓缺血再灌注损伤发生后均可见后肢功能逐渐好转现象,评分结果为缺血再灌注24 h达最高水平,48 h又略有下降;(2)免疫印迹结果:对照组PDIA3表达有清晰印迹显示,缺血再灌注0 h印迹轻度增强,6-12 h进一步增强,24 h显著减弱至最低水平,48 h再次升高到6-12 h水平;(3)免疫组织化学结果:神经元胞浆中可见PDIA3表达,且中间神经元表达量明显多于运动神经元;(4)结果表明:脊髓缺血再灌注损伤发生发展过程中PDIA3异常上调,说明PDIA3与脊髓缺血再灌注损伤密切相关,可作为其诊断和治疗的新靶点。
[Abstract]:Background: at present, it is believed that spinal cord ischemia-reperfusion injury is the main cause of secondary paralysis after decompression of spinal cord. The control of stress-related proteins and excitatory amino acids is very important in the treatment of spinal cord ischemia-reperfusion injury. Objective: to observe the expression of protein disulfide isomerase A _ 3 / PDIA _ 3 after spinal cord ischemia-reperfusion injury in rabbits. Rabbit spinal cord ischemia-reperfusion injury model was established by Zivin method. 36 New Zealand white rabbits were randomly divided into 6 groups. The control group only exposed the abdominal aorta without blocking the blood flow for 30 min and closed the abdominal cavity. In the experimental group, abdominal aorta blood flow was blocked for 30 min and the abdominal cavity was closed for 48 h after reperfusion. The motor function was evaluated by modified Tarlov score. Then the L3-L5 of the injured lumbar spinal cord was selected, then the PDIA3 was screened by fluorescence differential two dimensional gel electrophoresis and mass spectrometry, and then the mass spectrometry was confirmed by Western blotting. Finally, the changes of time and space expression in spinal cord were observed by immunohistochemical results. Results and conclusion: the results of motor function score showed that hindlimb work was observed in experimental animals after spinal cord ischemia-reperfusion injury. Can gradually improve the phenomenon, The results of Western blotting showed that the expression of PDIA3 in the control group was clearly imprinted, which reached the highest level at 24 h and decreased slightly at 48 h. The immunohistochemical results showed that the PDIA3 expression was observed in the cytoplasm of the neurons after 0 h ischemia reperfusion, slight enhancement at 6-12 h, and further enhancement at 24 h, significantly decreased to the lowest level at 48 h and increased again to the 6-12 h level (P < 0.05). The results showed that PDIA3 was upregulated during the development of spinal cord ischemia-reperfusion injury, indicating that PDIA3 was closely related to spinal cord ischemia-reperfusion injury. It can be used as a new target for diagnosis and treatment.
【作者单位】： 吉林大学第二医院骨科;
【基金】：国家自然科学基金(31572217,81350013) 吉林省科技型中小企业技术创新基金(SC201502001) 吉林大学研究生创新基金资助项目(2017176)~~
【分类号】：R-332;R651.2
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