缺血预处理对鼠后肢缺血再灌注脊髓运动神经元和下肢肌肉的保护作用

发布时间：2018-05-14 04:18

  本文选题：缺血预处理 + 缺血再灌注损伤　； 参考：《天津医科大学》2012年硕士论文

【摘要】：目的：缺血预处理(IPC)是以多次短暂缺血的方法提高组织或器官的缺血耐受性,减轻组织后续缺血再灌注(I／R)损伤。缺血预处理对心脏和骨骼肌等组织缺血再灌注损伤的保护作用已被实验证实,对周围神经缺血再灌注损伤的保护作用报道较少。而临床上周围神经缺血再灌注损伤有一定的发生率,考虑到缺血预处理的保护作用能在短时间内被诱导且方法简便易行,损伤副作用小,所以本实验重点研究缺血预处理对周围神经缺血再灌注损伤是否具有保护作用,通过建立大鼠一侧后肢的缺血再灌注损伤模型,进行不同条件缺血预处理干预,观察大鼠脊髓腰骶膨大段前角运动神经元以及胫前肌组织形态学、超微结构的变化,探讨缺血预处理对缺血再灌注损伤的保护作用,为临床有效防治此类损伤提供理论依据。 方法：健康3月龄SD大鼠64只,随机分为8组,每组8只,以缺血6h再灌注2h和12h分为A、B两个大组,按无预处理、预处理1次、预处理2次和预处理3次(叠加预处理时无时间间隔)分成A0、A1、A2、A3；B0、B1、B2、B38组, A0、B0为无缺血预处理的损伤对照组,其余为实验组。做左侧腹股沟切口,显露髂总、髂内和髂外动脉血管,以血管夹暂时夹闭阻断血流,建立一侧后肢缺血模型,预处理方式为夹闭血管阻断血流10min复流10min。取材部位为实验侧腰骶膨大段脊髓前角和胫前肌组织,放大镜下取材,行光镜HE染色及透射电镜醋酸铀和柠檬酸铅双重染色,观察不同条件缺血预处理大鼠一侧后肢在遭受缺血再灌注损伤时实验侧脊髓腰骶膨大段前角运动神经元以及后肢胫前肌组织形态学、超微结构变化。 结果： 1.大鼠周围神经缺血再灌注损伤动物模型制备成功,夹闭一侧髂总、髂内和髂外动脉可有效阻断一侧后肢动脉血流,放松血管夹后,又能使血供恢复,符合实验要求。 2.光镜下组织形态学观察结果：A0缺血6h再灌注2h组脊髓腰骶膨大段前角运动神经元数量减少,核溶解消失,损伤明显,预处理后神经元数量增多(P0.05),可见部分细胞核存在；B0缺血6h再灌注12h组神经元细胞大部分坏死溶解,预处理后坏死溶解区域减小。 3.电镜下超微结构观察：A0和B0组脊髓前角神经元细胞核周器不同程度扩张损伤,内质网扩张、大量线粒体空泡以及核膜溶解消失,B0组损伤最重。预处理后再灌注损伤程度有所减轻,叠加预处理后损伤进一步减轻。 4.骨骼肌虽通过不同条件预处理缺血再灌注后仍出现损伤表现,但损伤程度不同,相同预处理条件下的损伤程度随再灌注时间延长而加重。再灌注时间相同的情况下,增加预处理次数可相对减轻骨骼肌的再灌注损伤。 结论： 1.缺血预处理能减轻大鼠后肢缺血再灌注后腰骶膨大段脊髓前角运动神经元的损伤,从而具有保护作用。反复3次缺血预处理产生的保护作用稍优于2次预处理,3次、2次预处理的保护作用明显优于1次缺血预处理。 2.缺血预处理对骨髂肌同样有保护作用。2次预处理效果优于1次,2次、3次预处理相比较效果相似。同样预处理条件,在一定范围内时间,再灌注时间越长,损伤越严重。
[Abstract]:Objective: ischemic preconditioning (IPC) is a method to improve the ischemic tolerance of tissues or organs by multiple transient ischemia and reduce the injury of subsequent ischemia-reperfusion (I / R). The protective effect of ischemic preconditioning on ischemia-reperfusion injury of cardiac and skeletal muscles has been verified and the protection of ischemia reperfusion injury of peripheral nerve is made. There are few reports. There is a certain incidence of ischemic reperfusion injury in clinical peripheral nerve. It is considered that the protective effect of ischemic preconditioning can be induced in a short time and the method is simple and easy to use, and the damage side effect is small. Therefore, this experiment focuses on whether the ischemic preconditioning has protective effect on peripheral nerve ischemia reperfusion injury. The ischemia-reperfusion injury model of the hind limbs of the rat was established and the ischemic preconditioning intervention was carried out to observe the changes of the histomorphology and ultrastructure of the anterior horn of the lumbosacral lumbosacral segment of the spinal cord of the rat, and the protective effect of ischemic preconditioning on the ischemia reperfusion injury in order to effectively prevent and cure such injury in clinical. For theoretical basis.
Methods: 64 healthy 3 month old SD rats were randomly divided into 8 groups, with 8 rats in each group. With ischemic 6h reperfusion, 2h and 12h were divided into A, B, two large groups, which were pretreated 1 times without preconditioning, preconditioning 2 times and 3 preconditioning (preconditioning without time interval), A0, A1, A2, A3; B0, B1, A3, etc., the rest were the control group without ischemic preconditioning, and the remainder for the rest control group, and the rest for the rest of the control group without ischemic preconditioning. The rest were the rest of the control group, and the rest for the rest of the control group without ischemic preconditioning. The rest were the rest for the rest group, the rest were the rest control group without ischemic preconditioning. In the experimental group, the left inguinal incision was made to expose the blood vessels of the iliac and iliac internal and external iliac arteries. The blood flow was temporarily blocked by the clamp, and the ischemic model of one side hind limb was established. The preconditioning method was to clamp the blood flow of the 10min reflow 10min. to the anterior spinal cord and the anterior tibial muscle in the lumbosacral enlarged segment of the experimental side. HE staining and transmission electron microscope staining of uranium acetate and lead citrate were used to observe the histomorphology of the anterior horn of the lumbosacral anterior horn of the spinal cord and the posterior tibial muscle in the experimental side of the ischemic preconditioning rats and the ultrastructure of the posterior tibial muscle at the experimental side of ischemic preconditioning.
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