本文關鍵詞： 脊髓 缺血 再灌注損傷 蛋白質二硫鍵異構酶 組織構建 組織工程 脊髓缺血再灌注損傷 蛋白質二硫鍵異構酶A 應激 國家自然科學基金　出處：《中國組織工程研究》2017年24期 　論文類型：期刊論文

【摘要】：背景:目前認為脊髓缺血再灌注損傷是造成脊髓減壓手術后"二次癱瘓"的主要原因,而控制應激相關蛋白和興奮性氨基酸對脊髓缺血再灌注損傷的治療至關重要。目的:觀察蛋白質二硫鍵異構酶A3(PDIA3)在兔脊髓缺血再灌注損傷后表達變化。方法:依據Zivin法建立兔脊髓缺血再灌注損傷模型,將36只新西蘭白兔隨機均分成6組,對照組只顯露腹主動脈而不阻斷血流,30 min后關閉腹腔;實驗組阻斷腹主動脈血流30 min后分別再灌注0,6,12,24,48 h再關閉腹腔。首先均利用改良Tarlov評分進行運動功能評價;隨后取損傷段腰髓(L3-L5),結合熒光差異雙向凝膠電泳和質譜分析篩選出PDIA3;其次應用免疫印跡印證質譜;最后通過免疫組織化學結果觀察其在脊髓內的時間和空間表達變化特點。結果與結論:(1)運動功能評分結果:實驗動物于脊髓缺血再灌注損傷發(fā)生后均可見后肢功能逐漸好轉現象,評分結果為缺血再灌注24 h達最高水平,48 h又略有下降;(2)免疫印跡結果:對照組PDIA3表達有清晰印跡顯示,缺血再灌注0 h印跡輕度增強,6-12 h進一步增強,24 h顯著減弱至最低水平,48 h再次升高到6-12 h水平;(3)免疫組織化學結果:神經元胞漿中可見PDIA3表達,且中間神經元表達量明顯多于運動神經元;(4)結果表明:脊髓缺血再灌注損傷發(fā)生發(fā)展過程中PDIA3異常上調,說明PDIA3與脊髓缺血再灌注損傷密切相關,可作為其診斷和治療的新靶點。
[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.
【作者單位】： 吉林大學第二醫(yī)院骨科;
【基金】：國家自然科學基金(31572217,81350013) 吉林省科技型中小企業(yè)技術創(chuàng)新基金(SC201502001) 吉林大學研究生創(chuàng)新基金資助項目(2017176)~~
【分類號】：R-332;R651.2
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本文編號：1526420