孕酮对脊髓缺血再灌注损伤的修复作用及其机制研究
发布时间：2018-10-17 20:14
【摘要】：目的通过孕酮对兔脊髓缺血再灌注损伤模型的的干预,研究孕酮对兔脊髓缺血再灌注损伤的作用。方法采用腹主动脉阻断法诱导兔子的脊髓缺血再灌注模型,其中空白对照组3只仅进行手术不进行主动脉的阻断,随时间延长无明显病理变化;损伤组15只,手术后不给于药物干预;孕酮组15只,分别在术后0h、24h、48h腹腔注射8mg/kg孕酮。损伤组、孕酮组分别在术后12h、24h、36h、48h、72h各处死3只兔子。选取术后24h、48h及72h三个观察时间点,采用Tarlov评分对兔的后肢神经学功能进行评估。收集兔的脊髓组织样本,采用HE染色观察脊髓组织的形态变化,并对其神经元的保留水平进行统计。采用免疫组化的方法对脊髓组织中Caspase-8及p53蛋白的表达水平进行检测,并统计其中阳性细胞的数目。收集新鲜的组织样本采用western blotting技术对不同时间点的p53的蛋白表达情况进行检测,然后对其进行定量分析。结果1 HE染色显示,与空白对照组相比,损伤组12h无明显病变,24h脊髓出现出现空泡变性,神经元开始皱缩;48h神经元大量凋亡,组织出血水肿严重;72h出现炎性细胞浸润;孕酮组在术后24h到72h内也出现损伤组类似的变化,但神经元保存数量较多,且出血水肿及炎性细胞浸润等病理变化较轻。2术后12h时,空白对照组的神经元数量最多,孕酮组次之,损伤组最少;术后24h时,三组兔子的神经元数量无明显统计学差异(P0.05);术后48h时,损伤组及孕酮组的神经元数量与空白对照组相比明显减少(P0.05),且损伤组的神经元数量明显的少于孕酮组(P0.05);术后72h时,损伤组及孕酮组的神经元数量较空白对照组及术后24h和48h进一步减少(P0.05),且损伤组的神经元数量明显的小于孕酮组(P0.05)。3术后12h,损伤组及孕酮组兔子的神经功能均发生不同程度神经功能障碍,且损伤组的损伤程度更为严重(P0.05);手术48h后损伤组的运动功能较之前显著降低,孕酮组兔子较术后24h时有细微降低,但仍其功能显著高于损伤组(P0.05);手术72h时,损伤组及孕酮组兔子的运动神经功能均有不同程度的降低,但降低程度并无统计学差异,且孕酮组的评分显著高于损伤组(P0.05)。4三组兔子的脊髓组织的Caspase-8表达水平进行比较发现,12h时,空白对照组的Caspase-8的阳性得分最低,损伤组显著高于空白对照组,孕酮组的得分介于两组之间;48h时,损伤组及孕酮组的Caspase-8的蛋白表达水平进一步提高,且损伤组的表达水平显著高于孕酮组;72h时,损伤组的Caspase-8的表达水平较48h时进一步显著提高,孕酮组虽有提高,但提高幅度明显小于损伤组。5对p53的阳性细胞进行统计发现,24h时,空白对照组的p53阳性得分最低,损伤组为最高,孕酮组的得分介于两组之间;48h时,损伤组及孕酮组的p53的蛋白表达水平进一步提高,且损伤组的表达水平显著高于孕酮组;72h时,损伤组的p53的表达水平较48h时进一步显著提高,孕酮组虽有提高,但提高幅度明显小于损伤组。结论本研究中,通过对不同处理状态下脊髓缺血再灌注损伤兔子脊髓形态及炎症的进行评估,并对可能的机制进行初步的探索发现:1脊髓缺血再灌注损伤后注射孕酮能够显著的减少脊髓组织的炎症,降低脊髓神经元的凋亡;2脊髓缺血再灌注损伤后接受孕酮干预能够显著的提高兔子的运动神经功能的恢复;3脊髓缺血再灌注损伤后接受孕酮干预能够显著降低脊髓组织中Caspase-8及p53蛋白的表达,提示孕酮可能通过降低Caspase-8及p53蛋白的表达减少脊髓缺血损伤后神经元的凋亡。
[Abstract]:Objective To study the effect of progesterone on ischemia-reperfusion injury of spinal cord in rabbits by intervention of progesterone on rabbit spinal cord ischemia-reperfusion injury model. Methods The rabbit spinal cord ischemia/ reperfusion model was induced by abdominal aorta occlusion method, in which 3 groups were only operated without aortic occlusion, no obvious pathological changes were observed over time, 15 were injured in the injury group, no drug intervention was given after the operation, and 15 in the progesterone group. 8mg/ kg progesterone was injected intraperitoneally at 0h, 24h and 48h after operation respectively. Three rabbits were sacrificed at 12h, 24h, 36h, 48h and 72h after operation. The neurological function of the hindlimb of rabbits was evaluated using the lolov score at the three observation time points of 24h, 48h and 72h after operation. The spinal cord tissue samples of rabbits were collected, the morphological changes of spinal cord tissues were observed by HE staining and the retention levels of neurons were counted. The expression level of Caspase-8 and p53 protein in spinal cord tissue was detected by immunohistochemistry and the number of positive cells was counted. The expression of p53 protein in different time points was detected by western blotting technique and quantitative analysis was carried out. Results 1 HE staining showed that there were no obvious pathological changes in 12h of injury group compared with blank control group, vacuous degeneration occurred in 24h spinal cord, and neurons began to collapse; 48h neurons were apoptosis, and the edema of tissue hemorrhage was severe; 72h experienced inflammatory cell infiltration; There were similar changes in the group of progesterone in 24h to 721h after operation, but the number of neurons was much higher, and the pathological changes of hemorrhage edema and inflammatory cell infiltration were lighter. At 12h after operation, the number of neurons in the blank control group was the most, the progesterone group was the second, the injury group was the least, and when the operation was 24h, There was no significant difference in the number of neurons in the three groups (P0.05). When 48h after operation, the number of neurons in the injury group and progesterone group decreased significantly compared with the blank control group (P0.05), and the number of neurons in the injured group was significantly lower than that of the progesterone group (P0.05). The number of neurons in the injury group and progesterone group decreased further (P0.05), and the number of neurons in the injured group was significantly lower than that of the progesterone group (P0.05). The damage level of the injured group was more serious (P0.05), the exercise function of the injured group decreased significantly before the 48h operation, but the level of the progesterone group was significantly lower than that of the injured group at 24h, but its function was significantly higher than that of the injury group (P0.05). The level of Caspase-8 expression in the spinal cord tissue of the three groups of rabbits was significantly higher than that in the injured group (P0.05). The expression level of Caspase-8 in the injury group and progesterone group was higher than that in the control group, and the expression level of Caspase-8 in the injured group was significantly higher than that of the progesterone group at 48h, and the expression level of the injured group was significantly higher than that of the progesterone group. The expression level of Caspase-8 in the injured group was further improved when the expression level of Caspase-8 was higher than that in the injured group, but the increase in the progesterone group was significantly less than that of the injury group. The positive cells of p53 were statistically found to be the lowest in the blank control group and the highest in the injury group at 24h. The expression level of p53 in the injury group and the progesterone group was further increased after 48h, and the expression level of the injured group was significantly higher than that of the progesterone group at 48h. The expression level of p53 in the injured group was further improved when the expression level of p53 in the injured group was higher than that of the progesterone group at 48h. but the increase in amplitude is significantly less than that of the injury group. Conclusion In this study, the spinal cord morphology and inflammation of rabbits were assessed by ischemia-reperfusion injury of spinal cord in different treatment states, and the possible mechanisms were preliminarily explored. reducing the apoptosis of spinal cord neurons; receiving progesterone intervention after spinal ischemic reperfusion injury to remarkably improve the recovery of motor nerve function of rabbits; 3, receiving progesterone intervention after spinal ischemia reperfusion injury to remarkably reduce the expression of Caspase-8 and p53 protein in spinal cord tissues, It is suggested that progesterone may decrease the apoptosis of neurons after spinal ischemic injury by reducing the expression of Caspase-8 and p53 protein.
【学位授予单位】：华北理工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R651.2
本文编号：2277831
[Abstract]:Objective To study the effect of progesterone on ischemia-reperfusion injury of spinal cord in rabbits by intervention of progesterone on rabbit spinal cord ischemia-reperfusion injury model. Methods The rabbit spinal cord ischemia/ reperfusion model was induced by abdominal aorta occlusion method, in which 3 groups were only operated without aortic occlusion, no obvious pathological changes were observed over time, 15 were injured in the injury group, no drug intervention was given after the operation, and 15 in the progesterone group. 8mg/ kg progesterone was injected intraperitoneally at 0h, 24h and 48h after operation respectively. Three rabbits were sacrificed at 12h, 24h, 36h, 48h and 72h after operation. The neurological function of the hindlimb of rabbits was evaluated using the lolov score at the three observation time points of 24h, 48h and 72h after operation. The spinal cord tissue samples of rabbits were collected, the morphological changes of spinal cord tissues were observed by HE staining and the retention levels of neurons were counted. The expression level of Caspase-8 and p53 protein in spinal cord tissue was detected by immunohistochemistry and the number of positive cells was counted. The expression of p53 protein in different time points was detected by western blotting technique and quantitative analysis was carried out. Results 1 HE staining showed that there were no obvious pathological changes in 12h of injury group compared with blank control group, vacuous degeneration occurred in 24h spinal cord, and neurons began to collapse; 48h neurons were apoptosis, and the edema of tissue hemorrhage was severe; 72h experienced inflammatory cell infiltration; There were similar changes in the group of progesterone in 24h to 721h after operation, but the number of neurons was much higher, and the pathological changes of hemorrhage edema and inflammatory cell infiltration were lighter. At 12h after operation, the number of neurons in the blank control group was the most, the progesterone group was the second, the injury group was the least, and when the operation was 24h, There was no significant difference in the number of neurons in the three groups (P0.05). When 48h after operation, the number of neurons in the injury group and progesterone group decreased significantly compared with the blank control group (P0.05), and the number of neurons in the injured group was significantly lower than that of the progesterone group (P0.05). The number of neurons in the injury group and progesterone group decreased further (P0.05), and the number of neurons in the injured group was significantly lower than that of the progesterone group (P0.05). The damage level of the injured group was more serious (P0.05), the exercise function of the injured group decreased significantly before the 48h operation, but the level of the progesterone group was significantly lower than that of the injured group at 24h, but its function was significantly higher than that of the injury group (P0.05). The level of Caspase-8 expression in the spinal cord tissue of the three groups of rabbits was significantly higher than that in the injured group (P0.05). The expression level of Caspase-8 in the injury group and progesterone group was higher than that in the control group, and the expression level of Caspase-8 in the injured group was significantly higher than that of the progesterone group at 48h, and the expression level of the injured group was significantly higher than that of the progesterone group. The expression level of Caspase-8 in the injured group was further improved when the expression level of Caspase-8 was higher than that in the injured group, but the increase in the progesterone group was significantly less than that of the injury group. The positive cells of p53 were statistically found to be the lowest in the blank control group and the highest in the injury group at 24h. The expression level of p53 in the injury group and the progesterone group was further increased after 48h, and the expression level of the injured group was significantly higher than that of the progesterone group at 48h. The expression level of p53 in the injured group was further improved when the expression level of p53 in the injured group was higher than that of the progesterone group at 48h. but the increase in amplitude is significantly less than that of the injury group. Conclusion In this study, the spinal cord morphology and inflammation of rabbits were assessed by ischemia-reperfusion injury of spinal cord in different treatment states, and the possible mechanisms were preliminarily explored. reducing the apoptosis of spinal cord neurons; receiving progesterone intervention after spinal ischemic reperfusion injury to remarkably improve the recovery of motor nerve function of rabbits; 3, receiving progesterone intervention after spinal ischemia reperfusion injury to remarkably reduce the expression of Caspase-8 and p53 protein in spinal cord tissues, It is suggested that progesterone may decrease the apoptosis of neurons after spinal ischemic injury by reducing the expression of Caspase-8 and p53 protein.
【学位授予单位】：华北理工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R651.2
【参考文献】
相关期刊论文 前10条
1 李晓宁;吴磊;迟蕾;綦雪巍;梅继林;李诺;;不同治疗周期夹脊电针对急性脊髓损伤大鼠运动功能及细胞凋亡的影响[J];针刺研究;2016年06期
2 解立杰;黄锦秀;胡霁;;丙泊酚静脉泵注对兔脊髓缺血再灌注损伤的防治作用及其机制[J];山东医药;2016年43期
3 荆瀛黎;刘小野;白帆;董浩;陈惠;;褪黑素对脊髓损伤大鼠突触可塑性的作用[J];中国康复理论与实践;2016年07期
4 郑启新;田青;郭晓东;邓享誉;;脊髓损伤中坏死性凋亡的发生与调控机制研究现状及展望[J];中华实验外科杂志;2016年07期
5 余奇劲;解立杰;胡霁;;脊髓缺血再灌注损伤与血脊髓屏障关系的研究进展[J];实用医学杂志;2016年12期
6 吕少君;翁浩;刘海健;齐乐;徐志勇;刘荣;黄丁丁;;右美托咪定对兔脊髓缺血再灌注损伤促炎性因子表达的影响[J];中国临床药理学杂志;2016年06期
7 张芬;;MicroRNA-124a对SCI继发性损伤的影响及相关机制研究[J];郑州大学学报(理学版);2016年01期
8 祁磊;王茂德;;创伤性脑损伤与神经凋亡的研究进展[J];中华脑科疾病与康复杂志(电子版);2016年01期
9 赵文涛;刘明清;袁红兵;李盼盼;;神经节苷脂治疗急性脊髓损伤的疗效评价[J];山东医药;2015年39期
10 谢鹏;阮文辉;;骨髓间充质干细胞联合促红细胞生成素对脊髓损伤大鼠的保护作用[J];海南医学院学报;2016年01期
相关会议论文 前1条
1 任永信;殷国勇;张宁;;兔脊髓缺血再灌注损伤时Bcl-xL/Bcl-2相关死亡启动因子的变化及其意义[A];第三届全国脊髓损伤治疗与康复研讨会论文集[C];2012年
相关硕士学位论文 前1条
1 黄伟华;右美托咪定对家兔脊髓缺血再灌注损伤后Bax,Bcl-2,Caspase-3表达的影响[D];南方医科大学;2014年
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