普羅布考對血管性癡呆大鼠海馬組織SOD、MDA影響的實驗性研究
發(fā)布時間：2019-05-18 16:59
【摘要】：目的:血管性癡呆(vascular dementia, VD)是腦血管疾病的常見并發(fā)癥，它是由一系列腦血管因素導(dǎo)致腦組織損害引起的以認知功能障礙為特征的癡呆綜合癥，不僅給患者帶來長期痛苦，嚴重影響其生活質(zhì)量，而且給家庭、社會造成沉重負擔(dān)。近一個世紀(jì)，由于人類平均年齡的逐步增加，導(dǎo)致癡呆人群的不斷增加，特別是血管性癡呆，在歐洲被認為是繼阿爾茨海默病后導(dǎo)致癡呆的第二位因素，在亞洲可能占有的地位更為重要。因此對血管性癡呆的研究也就顯得特別重要。近年來，自由基損傷與血管性癡呆之間的關(guān)系逐漸成為人們研究的熱點問題。組織中丙二醛(MaleicDialdehyde, MDA)是脂質(zhì)過氧化物的主要代謝產(chǎn)物，超氧化物歧化酶(Superoxide Dismutase, SOD)活力的高低間接反映了機體清除氧自由基的能力。本研究旨在了解血管性癡呆大鼠海馬組織中SOD和MDA的變化，以進一步探討VD與氧化損傷的關(guān)系，并進一步探討普羅布考對VD模型大鼠智能改善的情況及海馬區(qū)SOD和MDA的影響，為臨床使用普羅布考治療VD提供實驗依據(jù)。 方法：選取健康雄性Sprague-Dawley (SD)大鼠，將大鼠隨機分為空白模型組、假手術(shù)組、普羅布考組，每組12只。對大鼠采用雙側(cè)頸總動脈永久性結(jié)扎的方法，造成慢性腦灌注不足所致VD模型。動物模型的篩選標(biāo)準(zhǔn)：術(shù)后7天在各組大鼠進行Morris水迷宮試驗訓(xùn)練1天，以假手術(shù)組大鼠平均逃避潛伏期的均值為參考值，計算手術(shù)組大鼠平均逃避潛伏期與參考值之差占平均逃避潛伏期的比例，該值20%定為癡呆鼠。普羅布考組給予普羅布考500mg/kg/d灌胃，假手術(shù)組與空白模型組給予等劑量羧甲基纖維素鈉灌胃。灌胃8周后，開始為期5天的Morris水迷宮實驗，前4天為定位航行試驗，記錄平均逃避潛伏期。第5天進行空間搜索試驗，記錄大鼠穿越原平臺次數(shù)。水迷宮實驗結(jié)束后，制作大腦海馬HE切片，采用光鏡觀察各組大鼠海馬CA1區(qū)的形態(tài)學(xué)變化；取海馬組織制成10%勻漿，采用黃嘌呤氧化酶法測定SOD活力，硫代巴比妥酸(Thibabituric Acid, TBA)法測MDA含量。通過觀察大鼠行為學(xué)、海馬組織形態(tài)學(xué)以及檢測海馬組織中SOD、MDA的表達，評價普羅布考對VD大鼠的作用效果。 結(jié)果： 1一般狀況觀察 早期觀察，手術(shù)后大鼠不能進食，運動減少，反應(yīng)遲鈍，但無肢體癱瘓。前6天體重逐漸減少，，每天降低5g左右，第7天大鼠體重開始恢復(fù)。上述觀察結(jié)果與文獻所描述的模型后表現(xiàn)大致相符，說明復(fù)制的模型成功。 2行為學(xué)測定 用Morris水迷宮觀察VD大鼠學(xué)習(xí)記憶情況，以大鼠逃避潛伏期代表學(xué)習(xí)能力，逃避潛伏期越短，學(xué)習(xí)能力越強。水迷宮訓(xùn)練的第1天起，空白模型組比假手術(shù)組的逃避潛伏期明顯延長(P0.01)，普羅布考組與空白模型組比較，逃避潛伏期明顯縮短(P0.01)。第5天空白模型組比假手術(shù)組的穿越平臺次數(shù)顯著減少(P0.01)。普羅布考組與空白模型組比較，穿越平臺次數(shù)明顯增加(P0.01)。各組大鼠隨著訓(xùn)練時間的推移，訓(xùn)練次數(shù)的增多，尋找平臺的方式在不斷變化，尋找平臺的時間都在縮短，說明學(xué)習(xí)鍛煉可以增強記憶能力。 3腦組織SOD活力和MDA含量的測定 空白模型組與假手術(shù)組相比，空白模型組SOD活性明顯降低，差異具有顯著性(P0.01)，MDA含量明顯增加(P0.01)；普羅布考組與空白模型組比較，普羅布考組SOD活性明顯增加(P0.01)，MDA含量明顯降低(P0.01)。 4海馬神經(jīng)細胞形態(tài)學(xué)觀察 假手術(shù)組海馬CA1區(qū)神經(jīng)細胞層結(jié)構(gòu)緊密，神經(jīng)細胞核完整，胞漿豐富�？瞻啄Ｐ徒M海馬CA1區(qū)的細胞排列紊亂,神經(jīng)細胞數(shù)明顯減少，許多細胞變得不完整，有的形成碎片,基質(zhì)疏松及微空泡形成。普羅布考組海馬CA1區(qū)的形狀與正常接近，大腦海馬區(qū)細胞排列緊密，層次豐富，細胞數(shù)明顯多于空白模型組，神經(jīng)細胞完整，胞漿豐富，可見少量變性細胞。 結(jié)論: 1大鼠雙側(cè)頸總動脈永久性結(jié)扎可以建立血管性癡呆模型。 2VD大鼠MDA含量明顯增加，SOD活性明顯降低，說明氧化損傷參與了VD的發(fā)病過程。 3普羅布考可以改善VD大鼠認知水平和病理變化，提高學(xué)習(xí)記憶能力。 4普羅布考可以提高VD大鼠大腦海馬中SOD活力，降低MDA含量，提示抗脂質(zhì)過氧化功能可能是普羅布考改善VD大鼠空間學(xué)習(xí)記憶障礙作用機制之一。
[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.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R965;R749.13
本文編號：2480177
[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.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R965;R749.13
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