赖氨酰氧化酶在大鼠脊髓慢性受压中的作用
发布时间：2019-07-08 09:43
【摘要】：目的通过渐进性螺钉植入法成功复制出慢性脊髓损伤的大鼠模型的基础上,比较赖氨酰氧化酶促进组、赖氨酰氧化酶抑制组、单纯慢性损伤压迫对照组各组间的不同,探讨赖氨酰氧化酶在大鼠脊髓慢性受压中的作用。观察不同组间皮层体感诱发电位、慢性脊髓损伤后组织结构变异的关系探讨赖氨酰氧化酶对于慢性脊髓压迫的治疗意义。方法健康SD雌性大鼠随机分为四组:空白对照组(A组,n=15);单纯慢性损伤压迫对照组(B组,n=15);压迫+BAPN LOX抑制组(C组,n=15);压迫+氯化钴LOX促进组(D组,n=15),建立慢性脊髓压迫模型后。B组,致伤后不做任何治疗处理,C组BAPN 1g/KG(腹腔注射),D组Co Cl2 30mg/L(给水)。饲养至8周以后,进行BBB评分、Tarlov运动评分及CSEP检测后获取大鼠脊髓部分组织进行RT-PCR检测赖氨酰氧化酶表达含量,部分组织制成石蜡包块儿进行HE及免疫组化染色。结果经脊髓组织RT-PCR检测结果显示与空白对照组A组相比各压迫组大鼠的脊髓内赖氨酰氧化酶的表达含量均有不同程度的升高。其中LOX促进组D组升高最为明显,这说明氯化钴成功的促进了赖氨酰氧化酶的表达。而LOX抑制组C组升高的程度最低,说明β氨基丙晴按照预期的设想抑制了赖氨酰氧化酶的表达。CSEP实验结果显示随时间变化除对照组A潜伏期未见变化外,其余各组潜伏期均出现不同程度的延长。测得各压迫组间结果在6周后出现差异,在8周差异明显。第8周时LOX促进组潜伏期为(18.06±0.67)ms,单纯慢性压迫阳性对照组为(19.76±0.82)ms,LOX抑制组为(22.16±0.74)ms。HE染色结果显示LOX抑制组D组与LOX促进组C组相比较白质中可见肿胀的轴突,轴突周围的空泡也较多,组织水肿较为严重、结构更为紊乱,细胞成分明显减少;灰质中可见神经元细胞皱缩更为明显,细胞核固缩,尼氏体更少甚至消失。利用免疫组化技术观察Bcl-2/Bax的表达情况从而检测脊髓神经细胞的凋亡情况。Bcl-2在各组中的表达水平为:LOX空白对照组A组的光密度值为(0.48±0.01),单纯慢性损伤压迫对照组B组的光密度值为(0.36±0.02),LOX抑制组C组的光密度值(0.25±0.02),LOX促进组D组的光密度值(0.31±0.01)。各组比较差异明显有统计学意义(P0.05)。Bax在空白对照组A组的表达最低,其余三组表达均升高,其中C组升高最为明显。D组较C组有所降低且差异有统计学意义(P0.05)。结论赖氨酰氧化酶可能在脊髓的慢性受压过程中起着重要的作用。
[Abstract]:Objective To study the model of rats with chronic spinal cord injury by means of progressive screw implantation. Objective To study the role of aminopeptidase in the chronic compression of spinal cord in rats. To observe the relationship between somatosensory evoked potential and chronic spinal cord injury in different groups, and to explore the significance of the method for the treatment of chronic spinal cord compression in the patients with chronic spinal cord injury. Methods Healthy SD female rats were randomly divided into four groups: blank control group (group A, n = 15), simple chronic injury compression control group (group B, n = 15), compression + BAPN LOX inhibition group (group C, n = 15), compression + cobalt chloride-type LOX promotion group (group D, n = 15), and establishment of chronic spinal cord compression model. Group B, no treatment after injury, group C BAPN 1g/ KG (intraperitoneal injection), and D group Co Cl2 30 mg/ L (water supply). After feeding for 8 weeks, BBB score, Tarlov's motion score and CSEP detection were performed to detect the expression level of the insulin-aminidase oxidase in the spinal cord of the rat, and the tissue was made into a paraffin-containing block for HE and immunohistochemical staining. Results The results of RT-PCR in the spinal cord showed that the level of the expression of the myeloperoxidase in the spinal cord of each compression group was higher than that of the control group A. In which LOX promoted the most significant increase in group D, suggesting that cobalt chloride successfully promoted the expression of the amine oxidase. While the level of the elevation of the group C in the LOX-inhibiting group was the lowest, and the inhibition of the expression of the aminopeptidase was inhibited according to the expected assumption. The results of CSEP showed that there was no change in the latency of the control group A, and the latency of the remaining groups was extended to a different extent. The difference in the results of each compression group after 6 weeks was measured, and the difference was evident at 8 weeks. At week 8, the latency of the LOX-promoting group was (18.06-0.67) ms, the simple chronic compression-positive control group was (19.76-0.82) ms, and the LOX-inhibiting group (22.16-0.74) ms. HE staining showed that the LOX-inhibiting group D and the LOX-promoting group C group compared the axons of the visible swelling in the white matter, The vacuoles around the axons are also more, the edema of the tissue is more serious, the structure is more disordered, the cell components are obviously reduced, the cells of the visible neurons in the gray matter are more obvious, the cells are fixed, and the ntroostite is less and even disappears. The expression of Bcl-2/ Bax was observed by immunohistochemistry in order to detect the apoptosis of spinal cord. The expression level of Bcl-2 in the control group was (0.48-0.01), the optical density value of group B was (0.36-0.02), and the optical density (0.25-0.02) in group C was inhibited in the control group. The optical density values of the group D were promoted by LOX (0.31-0.01). The expression of Bax in group A of blank control group was the lowest, and the expression of the other three groups increased, among which group C was the most obvious. There was a significant difference between group D and group C (P0.05). Conclusion The myeloperoxidase may play an important role in the chronic compression of the spinal cord.
【学位授予单位】：华北理工大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R651.2
本文编号：2511500
[Abstract]:Objective To study the model of rats with chronic spinal cord injury by means of progressive screw implantation. Objective To study the role of aminopeptidase in the chronic compression of spinal cord in rats. To observe the relationship between somatosensory evoked potential and chronic spinal cord injury in different groups, and to explore the significance of the method for the treatment of chronic spinal cord compression in the patients with chronic spinal cord injury. Methods Healthy SD female rats were randomly divided into four groups: blank control group (group A, n = 15), simple chronic injury compression control group (group B, n = 15), compression + BAPN LOX inhibition group (group C, n = 15), compression + cobalt chloride-type LOX promotion group (group D, n = 15), and establishment of chronic spinal cord compression model. Group B, no treatment after injury, group C BAPN 1g/ KG (intraperitoneal injection), and D group Co Cl2 30 mg/ L (water supply). After feeding for 8 weeks, BBB score, Tarlov's motion score and CSEP detection were performed to detect the expression level of the insulin-aminidase oxidase in the spinal cord of the rat, and the tissue was made into a paraffin-containing block for HE and immunohistochemical staining. Results The results of RT-PCR in the spinal cord showed that the level of the expression of the myeloperoxidase in the spinal cord of each compression group was higher than that of the control group A. In which LOX promoted the most significant increase in group D, suggesting that cobalt chloride successfully promoted the expression of the amine oxidase. While the level of the elevation of the group C in the LOX-inhibiting group was the lowest, and the inhibition of the expression of the aminopeptidase was inhibited according to the expected assumption. The results of CSEP showed that there was no change in the latency of the control group A, and the latency of the remaining groups was extended to a different extent. The difference in the results of each compression group after 6 weeks was measured, and the difference was evident at 8 weeks. At week 8, the latency of the LOX-promoting group was (18.06-0.67) ms, the simple chronic compression-positive control group was (19.76-0.82) ms, and the LOX-inhibiting group (22.16-0.74) ms. HE staining showed that the LOX-inhibiting group D and the LOX-promoting group C group compared the axons of the visible swelling in the white matter, The vacuoles around the axons are also more, the edema of the tissue is more serious, the structure is more disordered, the cell components are obviously reduced, the cells of the visible neurons in the gray matter are more obvious, the cells are fixed, and the ntroostite is less and even disappears. The expression of Bcl-2/ Bax was observed by immunohistochemistry in order to detect the apoptosis of spinal cord. The expression level of Bcl-2 in the control group was (0.48-0.01), the optical density value of group B was (0.36-0.02), and the optical density (0.25-0.02) in group C was inhibited in the control group. The optical density values of the group D were promoted by LOX (0.31-0.01). The expression of Bax in group A of blank control group was the lowest, and the expression of the other three groups increased, among which group C was the most obvious. There was a significant difference between group D and group C (P0.05). Conclusion The myeloperoxidase may play an important role in the chronic compression of the spinal cord.
【学位授予单位】：华北理工大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R651.2
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