Hemin诱导脑红蛋白高表达对脊髓损伤的保护作用

发布时间：2018-01-26 01:36

本文关键词： 氯化血红素脑红蛋白脊髓损伤大鼠　出处：《中国人民解放军医学院》2015年硕士论文　论文类型：学位论文

【摘要】：目的：制作大鼠Allen's脊髓损伤(Spinal Cord Injury, SCI)模型,腹腔注射氯化血红素(Hemin)溶液。分析Hemin干预脊髓损伤后大鼠行为学评分的变化和不同分组实验后一定时间点评分的差异,Hemin处理后大鼠脊髓内脑红蛋白(Neuroglobin, Ngb)表达空间和数量的规律,以及Hemin处理引起的脊髓Ngb高表达对于脊髓损伤后脊髓组织结构、细胞水平、病理进程影响,试为临床采用Hemin干预脊髓损伤提供可靠的实验室理论基础。方法：健康雄性SD大鼠90只,体重180g-200g,由军事医学科学院提供,随机分为三组：对照组(假手术组)、手术组(仅造模)、Hemin处理组(造模后腹腔注射Hemin溶液)。假手术组仅进行椎板卸下后还纳,不进行脊髓打击,造模后腹腔注射相同量生理盐水;手术组采用Allen's法制作大鼠损伤模型,造模后腹腔注射相同量生理盐水;Hemin处理组进行造模和相同量的Hemin溶液腹腔注射。采用大鼠后肢功能BBB评分、苏木素伊红染色、尼氏染色、免疫组化等方法评价各组大鼠实验后1d、3d、7d、14d、28d、56d时大鼠运动功能差异、脊髓结构改变、Ngb表达含量和分布的差异等。实验相关结果数据进行统计学分析。结果：(1)BBB评分结果：对照组Hemin处理组手术组。(2)HE染色结果：对照组脊髓组织完整,无明显的红细胞积聚及空隙,无炎性细胞浸润,神经细胞未出现病理改变;手术组脊髓具有明显的红细胞积聚及空隙,炎性细胞浸润明显,噬神经现象等病理改变严重;Hemin处理组损伤重于对照组,但明显轻于手术组,有少量出血和空腔,神经组织炎性细胞浸润现象较少,神经细胞水肿变性较少。(3)尼氏染色结果：对照组神经元形态正常,尼氏体围绕胞核呈颗粒状,颜色较深切且分布较多,由内向外颜色逐渐递减;Hemin处理组染色结果中水肿及颜色变浅、数量减少重于对照组但轻于手术组;手术组神经元病理变化最重,除上述情况外,更有尼氏体溶解的现象。(4)免疫组化结果：Hemin处理组阳性细胞最多,手术组次之,假手术组最少。结论：本实验通过采用Hemin腹腔注射干预脊髓损伤Allen's造模后的SD大鼠,对脊髓损伤引起脊髓组织Ngb高表达及Hemin诱导脊髓组织Ngb高表达的作用进行了验证。对Hemin诱导的脊髓组织Ngb高表达在脊髓损伤过程中的作用进行了研究和探讨。本实验的结果提示利用Hemin诱导脊髓Ngb高表达可以在提高大鼠对脊髓损伤的抗性。Ngb的神经保护作用涉及多途径、多环节,利用Hemin可以增强内源性Ngb的表达,从而提高其神经保护作用,这将为SCI的临床干预新的思路和药物选择提供可靠的实验室基础。
[Abstract]:Objective: to establish Allen's rat spinal cord injury (Spinal Cord, Injury, SCI) model, intraperitoneal injection of hemin (Hemin) solution. The difference analysis of the changes of Hemin intervention in rats after spinal cord injury behavior score and different experimental groups after a certain time point score, after the treatment of Hemin in spinal cord of rat Neuroglobin (Neuroglobin Ngb) expression space and number of rules, and Hemin induced high expression of Ngb in spinal cord tissue structure, cell level after spinal cord injury, pathological process, Hemin intervention of spinal cord injury provided theoretical basis for the reliable laboratory for clinical use. Methods: 90 healthy male SD rats, weight 180g-200g, from the Military Medical Science Academy of the PLA provide, randomly divided into three groups: control group (sham operation group), operation group (only model), Hemin group (model after intraperitoneal injection of Hemin solution). The sham operation group underwent laminectomy after discharge Accept, without spinal cord injury, rats after intraperitoneal injection of the same amount of normal saline; operation group by the method of Allen's injury model of rat model after intraperitoneal injection of the same amount of saline; Hemin treatment groups were modeled and the same amount of Hemin solution by intraperitoneal injection. Rat limb function after hematoxylin BBB score. With red staining, Nissl staining, immunohistochemistry methods were evaluated in rats after 1D, 3D, 7d, 14d, 28d, 56d between the motor function of rats, spinal cord structure change, the difference of Ngb expression content and distribution. The experimental results related data were statistically analyzed. Results: (1) results of BBB: control group Hemin group operation group. (2) the results of HE staining: control group of spinal cord tissue integrity, no accumulation of red cells and the gap is obvious, no infiltration of inflammatory cells, nerve cells did not appear pathological changes; spinal surgery group has obvious and accumulation of red blood cells Void, inflammatory cell infiltration was pathological changes serious neuronophagia phenomenon; Hemin treatment group injury in control group, but was lower in the operation group, a small amount of bleeding and cavity, less nerve tissue infiltration of inflammatory cells, nerve cell edema less. (3) Nissl staining results: the control group of neurons normal morphology, Nissl bodies around the nucleus is granular and color is deep and the distribution is more, from the inside to the outside color gradually decreased; Hemin staining results in the treatment group the edema and pale, heavier than the control group but lighter than the operation group to reduce the number of operation group; pathological changes of the neurons, in addition to the above, more chromatolysis phenomenon. (4) the results of immunohistochemistry: Hemin positive cells in treatment group, operation group and sham operation group at least. Conclusion: this experiment by using SD rats by intraperitoneal injection of Hemin intervention for spinal cord injury after Allen's, To cause the high expression of Ngb in spinal cord induced high expression of Ngb and Hemin in spinal cord injury was verified. On Hemin induced spinal cord tissue high expression of Ngb in spinal cord injury in rats was investigated and discussed. The experimental results suggest that the high expression of Ngb in rat spinal cord can improve the neuroprotective effect on rats the resistance of.Ngb spinal cord injury involving multi way, multi link using Hemin, using Hemin can enhance the expression of endogenous Ngb, so as to improve its neuroprotective effect, which will provide SCI clinical intervention ideas and new drugs provide reliable laboratory basis.

【学位授予单位】：中国人民解放军医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R651.2

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