普罗布考对血管性痴呆大鼠海马组织SOD、MDA影响的实验性研究

发布时间：2019-05-18 16:59

【摘要】：目的:血管性痴呆(vascular dementia, VD)是脑血管疾病的常见并发症，它是由一系列脑血管因素导致脑组织损害引起的以认知功能障碍为特征的痴呆综合症，不仅给患者带来长期痛苦，严重影响其生活质量，而且给家庭、社会造成沉重负担。近一个世纪，由于人类平均年龄的逐步增加，导致痴呆人群的不断增加，特别是血管性痴呆，在欧洲被认为是继阿尔茨海默病后导致痴呆的第二位因素，在亚洲可能占有的地位更为重要。因此对血管性痴呆的研究也就显得特别重要。近年来，自由基损伤与血管性痴呆之间的关系逐渐成为人们研究的热点问题。组织中丙二醛(MaleicDialdehyde, MDA)是脂质过氧化物的主要代谢产物，超氧化物歧化酶(Superoxide Dismutase, SOD)活力的高低间接反映了机体清除氧自由基的能力。本研究旨在了解血管性痴呆大鼠海马组织中SOD和MDA的变化，以进一步探讨VD与氧化损伤的关系，并进一步探讨普罗布考对VD模型大鼠智能改善的情况及海马区SOD和MDA的影响，为临床使用普罗布考治疗VD提供实验依据。方法：选取健康雄性Sprague-Dawley (SD)大鼠，将大鼠随机分为空白模型组、假手术组、普罗布考组，每组12只。对大鼠采用双侧颈总动脉永久性结扎的方法，造成慢性脑灌注不足所致VD模型。动物模型的筛选标准：术后7天在各组大鼠进行Morris水迷宫试验训练1天，以假手术组大鼠平均逃避潜伏期的均值为参考值，计算手术组大鼠平均逃避潜伏期与参考值之差占平均逃避潜伏期的比例，该值20%定为痴呆鼠。普罗布考组给予普罗布考500mg/kg/d灌胃，假手术组与空白模型组给予等剂量羧甲基纤维素钠灌胃。灌胃8周后，开始为期5天的Morris水迷宫实验，前4天为定位航行试验，记录平均逃避潜伏期。第5天进行空间搜索试验，记录大鼠穿越原平台次数。水迷宫实验结束后，制作大脑海马HE切片，采用光镜观察各组大鼠海马CA1区的形态学变化；取海马组织制成10%匀浆，采用黄嘌呤氧化酶法测定SOD活力，硫代巴比妥酸(Thibabituric Acid, TBA)法测MDA含量。通过观察大鼠行为学、海马组织形态学以及检测海马组织中SOD、MDA的表达，评价普罗布考对VD大鼠的作用效果。结果： 1一般状况观察早期观察，手术后大鼠不能进食，运动减少，反应迟钝，但无肢体瘫痪。前6天体重逐渐减少，，每天降低5g左右，第7天大鼠体重开始恢复。上述观察结果与文献所描述的模型后表现大致相符，说明复制的模型成功。 2行为学测定用Morris水迷宫观察VD大鼠学习记忆情况，以大鼠逃避潜伏期代表学习能力，逃避潜伏期越短，学习能力越强。水迷宫训练的第1天起，空白模型组比假手术组的逃避潜伏期明显延长(P0.01)，普罗布考组与空白模型组比较，逃避潜伏期明显缩短(P0.01)。第5天空白模型组比假手术组的穿越平台次数显著减少(P0.01)。普罗布考组与空白模型组比较，穿越平台次数明显增加(P0.01)。各组大鼠随着训练时间的推移，训练次数的增多，寻找平台的方式在不断变化，寻找平台的时间都在缩短，说明学习锻炼可以增强记忆能力。 3脑组织SOD活力和MDA含量的测定空白模型组与假手术组相比，空白模型组SOD活性明显降低，差异具有显著性(P0.01)，MDA含量明显增加(P0.01)；普罗布考组与空白模型组比较，普罗布考组SOD活性明显增加(P0.01)，MDA含量明显降低(P0.01)。 4海马神经细胞形态学观察假手术组海马CA1区神经细胞层结构紧密，神经细胞核完整，胞浆丰富。空白模型组海马CA1区的细胞排列紊乱,神经细胞数明显减少，许多细胞变得不完整，有的形成碎片,基质疏松及微空泡形成。普罗布考组海马CA1区的形状与正常接近，大脑海马区细胞排列紧密，层次丰富，细胞数明显多于空白模型组，神经细胞完整，胞浆丰富，可见少量变性细胞。结论: 1大鼠双侧颈总动脉永久性结扎可以建立血管性痴呆模型。 2VD大鼠MDA含量明显增加，SOD活性明显降低，说明氧化损伤参与了VD的发病过程。 3普罗布考可以改善VD大鼠认知水平和病理变化，提高学习记忆能力。 4普罗布考可以提高VD大鼠大脑海马中SOD活力，降低MDA含量，提示抗脂质过氧化功能可能是普罗布考改善VD大鼠空间学习记忆障碍作用机制之一。
[Abstract]:Objective: Vascular dementia (VD) is a common complication of cerebrovascular disease. It is a kind of dementia syndrome characterized by a series of cerebrovascular factors that cause brain tissue damage, which not only brings long-term pain to the patient, but also seriously affects the quality of life. And a heavy burden on the family and society. As a result of the gradual increase in the average age of humans in the last century, the increasing number of people with dementia, especially vascular dementia, is considered to be the second factor in dementia following the onset of Alzheimer's disease, which is more important in Asia. The study of vascular dementia is therefore of particular importance. In recent years, the relationship between free-radical injury and vascular dementia has become a hot topic of people's research. MaleicDialdehyde (MDA) in the tissue is the main metabolite of lipid peroxide, and the activity of superoxide dismutase (SOD) indirectly reflects the ability of the body to remove oxygen free radicals. The purpose of this study was to investigate the changes of SOD and MDA in the hippocampus of vascular dementia rats, to further explore the relationship between VD and oxidative damage, and to further study the effect of probucol on the intelligent improvement of VD model rats and the effects of SOD and MDA in the hippocampus. The experimental basis was provided for clinical use of probucol for the treatment of VD. Methods: Male Sprague-Dawley (SD) rats were randomly divided into blank model group, sham operation group and probucol group. The method of permanent ligation of the bilateral common carotid artery in rats was used to cause the VD model due to the deficiency of the chronic cerebral perfusion. Model of animal model:7 days after operation, Morris water maze test was performed for 1 day in each group of rats. The mean value of the average escape latency of the rats in the sham operation group was the reference value, and the difference between the average escape latency and the reference value of the operation group was calculated as the ratio of the average escape latency. In the case of dementia, the value is 20%. Rats. Probucol 500 mg/ kg/ day given probucol 500 mg/ kg/ day, and the sham operation group and blank model group were given equal doses of methyl cellulose sodium. After 8 weeks of gastric administration, Morris water maze test was started for 5 days. The first four days were used to position the navigation test and the average escape latency was recorded. The space search test was performed on the 5th day, and the rats were recorded through the original platform. The results were as follows: After the water maze test, the HE section of the brain of the brain was made, and the morphological changes of the hippocampal CA1 area of each group were observed by light microscope. The hippocampal formation was made into 10% homogenate, and the activity of SOD and thiobarbituric acid (TBA) were determined by the method of the method of the yellow-methotrexate oxidase. The effects of probucol on VD rats were evaluated by observing the behavior of rats, the morphology of the hippocampus and the detection of the expression of SOD and MDA in the hippocampus. Fruit. Results:1 The general situation was observed in the early stage. After the operation, the rats were not allowed to eat, the exercise was reduced, the reaction was slow, but no limb paralysis. The weight of the first six days is gradually reduced, about 5 grams per day, and the seventh day is large The body weight of the mouse is recovered. The results of the above observations are approximately in line with the post-model performance described in the literature and will be described the model of replication is Work. The behavior measurement was used to observe the learning and memory of VD rats by the Morris water maze, to avoid the latent period of the rats to represent the learning ability and to avoid the latent period. The shorter the latent period, the stronger the learning ability. In the first day of the water maze training, the escape latency of the blank model group compared with the sham operation group was significantly prolonged (P0.01), and the probucol group was compared with the blank model group to avoid the incubation period. Obvious shortening (P0.01). The number of cross platforms of the blank model group on the 5th day was lower than that of the sham-operated group. Significant decrease (P0.01). The number of crossing platforms is compared with the blank model group. As the training time, the number of training times increased, the way to find the platform is changing constantly, and the time of searching for the platform is shortened and explained. exercise can enhance memory. Brain tissue The SOD activity and MDA content in the blank model group were significantly lower than those in the sham operation group (P0.01), and the content of MDA was significantly increased (P0.01). Compared with the blank model group, the activity of SOD in probucol group was significantly increased (P0.01), and M. The content of DA was significantly lower (P0. 01).4. Morphological observation of the hippocampal neurons in the hippocampus of the hippocampus. The cell structure is tight, the nucleus of the nerve cell is complete and the cytoplasm is abundant. The cell arrangement of the hippocampal CA1 region of the blank model group is disordered, the number of the nerve cells is obviously reduced, and many of the cells become incomplete. The shape of the CA1 region of the hippocampus of the probucol group is close to that of the normal, the cells in the hippocampus of the brain are closely arranged, the level is abundant, the number of cells is obviously higher than that of the blank model group, transcell Complete, rich in cytoplasm and a small amount of denatured cells. Conclusion:1 The model of vascular dementia can be established by the permanent ligation of the bilateral common carotid artery of the rat. The content of MDA in the 2VD rats is obviously increased, and S OD activity was significantly reduced, indicating that oxidative damage was involved in the pathogenesis of VD. The study can improve the cognitive level and pathological changes of VD rats, and improve the learning and memory ability. Probucol can improve the activity of SOD in the hippocampus of VD rats, decrease the content of MDA, and prompt the anti-lipid peroxidation.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R965;R749.13

【参考文献】