豐富環(huán)境改善血管性癡呆大鼠學習和空間記憶能力及其機制研究
發(fā)布時間：2019-06-14 09:43
【摘要】：背景與目的:近年來由于人民群眾生活質(zhì)量的改善,心腦血管疾病的發(fā)生率也逐漸升高,由此導致血管性癡呆(VaD)的發(fā)病率也隨之上升。目前在所有癡呆類型中,VaD發(fā)病率排名第二,僅低于阿爾茨海默病(AD),它在給病人帶來生活上的困擾之外,也給社會和家庭造成了沉重的負擔,目前臨床上關(guān)于Va D病人的治療方法仍十分有限。近年來,對于豐富環(huán)境(EE)的研究日益增多,發(fā)現(xiàn)其能增強神經(jīng)的可塑性,在多種神經(jīng)精神疾病中已經(jīng)被證實有一定的治療作用。在本研究中,我們擬探討EE的干預是否能夠提升Va D大鼠的學習及空間記憶能力以及其可能的內(nèi)在作用機制。方法:本實驗采用雙側(cè)頸總動脈結(jié)扎法(Bilateral common carotid occlusion,2VO)建立SD(Sprague-Dawley)大鼠的VaD模型。采用健康成年的雄性SD大鼠,隨機分為4個組:假手術(shù)(sham)組、2VO組、假手術(shù)+豐富環(huán)境(sham+EE)組和2VO+豐富環(huán)境(2VO+EE)組。2VO組和2VO+EE組的大鼠進行2VO手術(shù)。有兩種不同的飼養(yǎng)環(huán)境在本實驗中被運用:標準飼養(yǎng)環(huán)境(SE)和豐富環(huán)境(EE)。Sham組和2VO組的大鼠在SE中飼養(yǎng)4周,而sham+EE組和2VO+EE組的大鼠則在EE中飼養(yǎng)4周。通過Morris水迷宮(MWM)來檢測大鼠的學習及空間記憶能力的差異,尼氏染色觀察大鼠神經(jīng)元的存活數(shù)量及形態(tài)學的改變,tunel法測定各組大鼠神經(jīng)細胞的凋亡情況,采用westernblot檢測wnt/?-catenin通路中wnt3a、p-gsk-3?、?-catenin的蛋白含量的變化。結(jié)果:(1)在mwm的實驗結(jié)果中,在ee中飼養(yǎng)的2vo+ee組大鼠的逃避潛伏期較se中飼養(yǎng)2vo組大鼠明顯縮短(p0.05),目標象限停留時間明顯增加(p0.05),在ee中飼養(yǎng)的sham+ee組大鼠逃避潛伏期較se中飼養(yǎng)的sham組大鼠明顯縮短(p0.05),目標象限停留時間明顯增加(p0.05),提示ee可以提高大鼠的學習和空間記憶能力。(2)與sham組相比,2vo組大鼠在術(shù)后4周的海馬凋亡細胞比例明顯增加(p0.05),而經(jīng)過ee干預的2vo+ee組的海馬凋亡細胞的比例則比2vo組明顯下降(p0.05),同時經(jīng)過ee干預的sham+ee組的海馬凋亡細胞比例也較se中飼養(yǎng)的sham組大鼠明顯下降(p0.05)。(3)與sham組相比,2vo組大鼠在術(shù)后4周的海馬正常存活的神經(jīng)元數(shù)量明顯減少(p0.05)。而2vo+ee組在經(jīng)過ee的干預后存活的神經(jīng)元數(shù)量則2vo組增加(p0.05),經(jīng)過ee干預的sham+ee組大鼠的海馬存活神經(jīng)元數(shù)量也明顯要高于飼養(yǎng)在se環(huán)境中的sham組(p0.05)。(5)westernblot的結(jié)果顯示,與se中飼養(yǎng)4周的sham組及2vo組大鼠相比,ee中飼養(yǎng)4周的sham+ee組和2vo+ee組大鼠海馬中wnt3a、p-gsk-3?、?-catenin蛋白的含量明顯增加(p0.05),提示大鼠在經(jīng)歷豐富環(huán)境的過程中伴隨著wnt/?-catenin信號通路的激活。結(jié)論:與SE中飼養(yǎng)的大鼠相比,經(jīng)歷4周EE的大鼠學習和空間記憶能力得到了明顯的提升。大鼠在EE的飼養(yǎng)環(huán)境下,海馬神經(jīng)元凋亡比例降低,正常神經(jīng)元的數(shù)量增加。在EE的作用過程中同時伴隨著wnt3a、p-GSK-3?、?-catenin蛋白含量的增加,說明4周的EE干預激活了wnt/?-catenin信號通路,提示wnt/?-catenin信號通路的激活可能是EE促進學習和空間記憶能力提升作用的內(nèi)在分子學機制之一。
[Abstract]:BACKGROUND & OBJECTIVE: In recent years, the incidence of cardiovascular and cerebrovascular diseases has increased gradually due to the improvement of the quality of people's life, and the incidence of vascular dementia (VaD) has also increased. In all the types of dementia, the rate of VaD is the second and only lower than the Alzheimer's disease (AD), which causes a heavy burden on the society and the family, which is still very limited in the treatment of the Va-D patients. In recent years, the research on the rich environment (EE) has been increasing, and it has been found that it can enhance the plasticity of the nerve, and has been proved to have a certain therapeutic effect in various neuropsychiatric diseases. In this study, we intend to explore whether the intervention of EE can improve the learning and spatial memory capacity of the Va-D rats and its possible internal mechanism of action. Methods: The VaD model of SD (Sprague-Dawley) rats was established by the double-sided common carotid artery ligation (2VO). Healthy adult male SD rats were randomly divided into 4 groups: sham group, 2VO group, sham operation + rich environment (sham + EE) group and 2VO + rich environment (2VO + EE) group. Two different breeding environments were used in this experiment: the standard feeding environment (SE) and the rich environment (EE). Rats in the Sham group and the 2 VO group were fed in SE for 4 weeks, while the sham + EE group and the 2 VO + EE group rats were housed in the EE for 4 weeks. The learning and spatial memory abilities of the rats were detected by the Morris water maze (MWM). The number of survival and the morphological changes of the neurons in the rats were observed by means of the method of Morris water maze (MWM), and the apoptosis of the nerve cells in each group was determined by means of the test. The wnt3a, p-gsk-3 in the wnt/?-cattenin pathway were detected by using the method of western blot. ? -a change in the protein content of catenin. Results: (1) In the experimental results of mwm, the escape latency of the 2 vo + ee group rats fed in ee was significantly shortened (p0.05), and the retention time of the target quadrant was significantly increased (p0.05). In the sham + ee group, the rats of sham + ee group were significantly shorter (p0.05) than that of the sham group, and the retention time of the target quadrant was significantly increased (p0.05), and it was suggested that the ee could improve the learning and spatial memory of the rats. (2) Compared with the sham group, the proportion of the apoptosis cells in the hippocampus of the 2vo + ee group after the operation was significantly increased (p0.05), while the proportion of the apoptotic cells in the hippocampus of the 2vo + ee group after the ee intervention was significantly lower than that of the second group (p0.05). At the same time, the percentage of apoptotic cells in the hippocampus of the sham + ee group by ee intervention was lower than that in the sham group (p0.05). (3) Compared with the sham group, the number of neurons in the 2 vo group in the normal survival of the hippocampus at 4 weeks after operation was significantly reduced (p0.05). The number of surviving neurons in the 2vo + ee group after the ee intervention increased (p0.05), and the number of surviving neurons in the hippocampus of the sham + ee group after the ee intervention was significantly higher than that in the sham group (p0.05) in the se environment. (5) The results of western blot showed that wnt3a, p-gsk-3 in the hippocampus of the 2 vo + ee group and the 2 vo + ee group were fed with 4 weeks of sham + ee and 2 vo + ee groups in the ee. ? The content of the-catenin protein was significantly increased (p0.05), suggesting that the rats were associated with the activation of the wnt/?-cattenin signal pathway in a rich environment. Conclusion: Compared with the rats fed with SE, the learning and spatial memory ability of rats subjected to 4 weeks of EE is obviously improved. In the environment of EE, the percentage of neuron apoptosis in the hippocampus decreased and the number of normal neurons increased. At the same time with wnt3a, p-GSK-3 in the course of the action of EE? ? The increase in the content of the-catenin protein suggests that the 4-week EE intervention has activated the wnt/?-cattenin signal pathway, suggesting that the activation of the wnt/?-catenin signal pathway may be one of the intrinsic molecular mechanisms of EE for learning and the enhancement of spatial memory capacity.
【學位授予單位】：重慶醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R749.1
本文編號：2499299
[Abstract]:BACKGROUND & OBJECTIVE: In recent years, the incidence of cardiovascular and cerebrovascular diseases has increased gradually due to the improvement of the quality of people's life, and the incidence of vascular dementia (VaD) has also increased. In all the types of dementia, the rate of VaD is the second and only lower than the Alzheimer's disease (AD), which causes a heavy burden on the society and the family, which is still very limited in the treatment of the Va-D patients. In recent years, the research on the rich environment (EE) has been increasing, and it has been found that it can enhance the plasticity of the nerve, and has been proved to have a certain therapeutic effect in various neuropsychiatric diseases. In this study, we intend to explore whether the intervention of EE can improve the learning and spatial memory capacity of the Va-D rats and its possible internal mechanism of action. Methods: The VaD model of SD (Sprague-Dawley) rats was established by the double-sided common carotid artery ligation (2VO). Healthy adult male SD rats were randomly divided into 4 groups: sham group, 2VO group, sham operation + rich environment (sham + EE) group and 2VO + rich environment (2VO + EE) group. Two different breeding environments were used in this experiment: the standard feeding environment (SE) and the rich environment (EE). Rats in the Sham group and the 2 VO group were fed in SE for 4 weeks, while the sham + EE group and the 2 VO + EE group rats were housed in the EE for 4 weeks. The learning and spatial memory abilities of the rats were detected by the Morris water maze (MWM). The number of survival and the morphological changes of the neurons in the rats were observed by means of the method of Morris water maze (MWM), and the apoptosis of the nerve cells in each group was determined by means of the test. The wnt3a, p-gsk-3 in the wnt/?-cattenin pathway were detected by using the method of western blot. ? -a change in the protein content of catenin. Results: (1) In the experimental results of mwm, the escape latency of the 2 vo + ee group rats fed in ee was significantly shortened (p0.05), and the retention time of the target quadrant was significantly increased (p0.05). In the sham + ee group, the rats of sham + ee group were significantly shorter (p0.05) than that of the sham group, and the retention time of the target quadrant was significantly increased (p0.05), and it was suggested that the ee could improve the learning and spatial memory of the rats. (2) Compared with the sham group, the proportion of the apoptosis cells in the hippocampus of the 2vo + ee group after the operation was significantly increased (p0.05), while the proportion of the apoptotic cells in the hippocampus of the 2vo + ee group after the ee intervention was significantly lower than that of the second group (p0.05). At the same time, the percentage of apoptotic cells in the hippocampus of the sham + ee group by ee intervention was lower than that in the sham group (p0.05). (3) Compared with the sham group, the number of neurons in the 2 vo group in the normal survival of the hippocampus at 4 weeks after operation was significantly reduced (p0.05). The number of surviving neurons in the 2vo + ee group after the ee intervention increased (p0.05), and the number of surviving neurons in the hippocampus of the sham + ee group after the ee intervention was significantly higher than that in the sham group (p0.05) in the se environment. (5) The results of western blot showed that wnt3a, p-gsk-3 in the hippocampus of the 2 vo + ee group and the 2 vo + ee group were fed with 4 weeks of sham + ee and 2 vo + ee groups in the ee. ? The content of the-catenin protein was significantly increased (p0.05), suggesting that the rats were associated with the activation of the wnt/?-cattenin signal pathway in a rich environment. Conclusion: Compared with the rats fed with SE, the learning and spatial memory ability of rats subjected to 4 weeks of EE is obviously improved. In the environment of EE, the percentage of neuron apoptosis in the hippocampus decreased and the number of normal neurons increased. At the same time with wnt3a, p-GSK-3 in the course of the action of EE? ? The increase in the content of the-catenin protein suggests that the 4-week EE intervention has activated the wnt/?-cattenin signal pathway, suggesting that the activation of the wnt/?-catenin signal pathway may be one of the intrinsic molecular mechanisms of EE for learning and the enhancement of spatial memory capacity.
【學位授予單位】：重慶醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R749.1
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