PSD大鼠小脑顶核形态学改变及其损伤机制研究

发布时间：2018-01-19 12:08

本文关键词： 卒中后抑郁小脑顶核凋亡形态学大鼠　出处：《辽宁医学院》2014年硕士论文　论文类型：学位论文

【摘要】：目的观察卒中后抑郁(post-stroke depression，PSD)模型大鼠小脑顶核细胞的形态学改变，并检测顶核细胞的凋亡情况。通过观察凋亡相关蛋白caspase-3、caspase-8、caspase-9及AIF（apoptosis-inducing factor，AIF）在该区域的表达，初步探讨PSD大鼠小脑顶核细胞发生损伤的方式，探讨小脑顶核损伤与PSD的关系。方法选用健康SD（Sprague-Dawley, SD）大鼠，随机分为4组。对照组：给予假手术处理。卒中组：行大脑中动脉闭塞术（middle cerebral artery occlusion，MCAO）。抑郁组：慢性不可预见性温和刺激（chronic unpredictable mildstimulation, CUMS）结合孤养。PSD组：MCAO后给予CUMS结合孤养。进行行为学观察和测试后，取脑组织，行尼氏染色，观察各组大鼠顶核区域形态学改变；应用末端脱氧核苷酰基转移酶介导性dUTP切口末端标记(Terminaldeoxynucleotidyl transferase-mediated dUTP nick end labeling, TUNEL)检测神经细胞凋亡情况，应用免疫组化技术测定凋亡相关蛋白caspase-3、caspase-8、caspase-9及AIF的表达。结果 1、大鼠行为学观察和测试：与对照组及卒中组相比，PSD组大鼠体重增长速度减慢，糖水消耗量下降，水平运动及垂直运动降低，强迫游泳不动时间延长（P0.05，或P0.01）。 2、脑组织病理学观察：尼氏染色结果，对照组尼氏小体正常存在。与对照组相比，PSD组小脑顶核细胞尼氏小体减少甚至消失。尼氏体在其余各组中均有减少，但程度不同。 3、TUNEL检测：对照组小脑顶核可见极少量的凋亡细胞，考虑与生理性死亡有关。卒中组，，抑郁组和PSD组均有较多凋亡细胞出现，与对照组相比具有统计学意义（P0.05，或P0.01）。 4、免疫组化检测结果：对照组caspase-3、caspase-8、caspase-9及AIF的表达强度均最低。与对照组比较，PSD组的caspase-3、caspase-8、caspase-9及AIF的表达均增加（P0.01）。结论 1、PSD大鼠小脑顶核细胞存在损伤，该损伤可能与PSD发病有关。 2、PSD大鼠中小脑顶核的损伤与凋亡相关，可能通过caspase及非caspase介导的通路传导，但具体的传导通路未确定。
[Abstract]:Purpose Objective: to observe the morphological changes of cerebellar parietal nucleus cells in post-stroke PSD rats with post-stroke depression. The apoptosis of apical nucleus cells was detected. The apoptosis-related protein caspase-3 and caspase-8 were observed. The expression of caspase-9 and AIF(apoptosis-inducing factor AIFs in this region. To explore the damage mode of cerebellar parietal nucleus cells in PSD rats and the relationship between the injury of cerebellar parietal nucleus and PSD. Method Healthy SD rats were selected. They were randomly divided into 4 groups: control group: sham operation and stroke group: middle cerebral artery occlusion was performed during middle cerebral artery occlusion (MCAO). Depression group: chronic unpredictable mild stimulation of chronic unpredictable mildstimulation. CUMS combined with solitary. PSD group was given CUMS combined with solitary care. After behavioral observation and test, brain tissue was taken and stained by Nissl's staining. The morphologic changes of the parietal nucleus in each group were observed. Terminal deoxynucleotidyl transferase mediated dUTP incision end labeling (. Terminaldeoxynucleotidyl transferase-mediated dUTP nick end labeling. Tunel was used to detect neuronal apoptosis and the apoptosis-related protein caspase-3 and caspase-8 were detected by immunohistochemistry. Expression of caspase-9 and AIF. Results 1. Behavioral observation and test: compared with the control group and the stroke group, the rats in the PSD group had slower weight gain, lower consumption of sugar water, and lower horizontal and vertical movement. Forced swimming immobility prolonged P0.05, or P0.01. 2. Observation of brain histopathology: the results of Nissl staining showed that the Nissl corpuscles were normal in the control group, and compared with the control group. In PSD group, the Nissl corpuscles of cerebellar parietal nucleus cells decreased or even disappeared, and the Nissl bodies decreased in other groups, but to different degrees. 3Tunel detection: a very small number of apoptotic cells were found in the cerebellar parietal nucleus in the control group, which was related to physiological death. There were more apoptotic cells in the apoplexy group, depression group and PSD group. Compared with the control group, there was significant difference in P0.05, or P0.01. 4. The expression of caspase-3, caspase-8, caspase-9 and AIF were the lowest in the control group. The expression of caspase-3, caspase-8, caspase-9 and AIF increased in PSD group (P 0.01). Conclusion 1 the damage of cerebellar parietal nucleus cells in PSD rats may be related to the pathogenesis of PSD. 2 the damage of cerebellar parietal nucleus is related to apoptosis, which may be mediated by caspase and non-#en1# pathway, but the specific pathway is not determined.
【学位授予单位】：辽宁医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R743.3

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