本文選題：阿爾茨海默病 + 血腦屏障��； 參考：《北京中醫(yī)藥大學》2017年博士論文

【摘要】：1研究目的阿爾茨海默病(Alzheimer' s Disease,AD)又稱老年癡呆,是一種以進行性認知障礙和記憶能力損害為主的中樞神經(jīng)系統(tǒng)退變性疾病。阿爾茨海默病是當前公共健康所面臨的重大問題,已被確定為中樞神經(jīng)系統(tǒng)損傷領域研究的重點疾病。因此研究老年性癡呆及其相關疾病已成為了刻不容緩的迫切任務,成為國內(nèi)外醫(yī)學研究的重點課題之一。臨床研究顯示,針灸可以改善AD患者的精神行為學狀態(tài)和認知功能。電針是一種可以針對多種疾病簡單有效的治療手段,許多的研究和證據(jù)表明針刺可以有效地改善AD,但其機制尚不明確,需要進一步探討。Aβ級聯(lián)假說認為β淀粉樣蛋白(β Amyloid protein,Aβ)的生成與清除失衡是其沉積于腦組織繼而形成淀粉樣斑塊的原因,其可能具有的神經(jīng)毒性作用引發(fā)神經(jīng)元變性,導致AD癥狀的產(chǎn)生。目前,也已經(jīng)有許多團隊從多個角度研究并證明其確實存在神經(jīng)毒性,Aβ在神經(jīng)的退化和凋亡過程中扮演著十分重要的角色。本課題組以電針"百會、涌泉"為治療手段,研究電針對不同月齡APP/PS1轉(zhuǎn)基因小鼠行為學的影響,探討電針治療阿爾茲海默病的最佳干預時間;基于A0的血腦屏障清除途徑,觀察電針對的APP/PS1轉(zhuǎn)基因AD模型腦內(nèi)和血清Aβ1-40和Aβ 1-42的表達,探討電針是否通過調(diào)節(jié)APP/PS1雙轉(zhuǎn)基因鼠低密度脂蛋白受體蛋白-1(LRP1)相關基因的表達,促進海馬內(nèi)β淀粉樣蛋白清除,探討電針治療阿爾茲海默病的機制。2研究方法首先,將4、6、9月齡APP/PS1雙轉(zhuǎn)基因小鼠各20只,隨機分為模型組、電針治療組,以相同月齡C57BL/6野生鼠各10只作為正常對照組;電針干預6周后,采用Morris水迷宮進行空間記憶行為學測試,觀察各月齡各組行為學的變化。之后,將6月齡APP/PS1雙轉(zhuǎn)基因小鼠64只,隨機分為模型組、電針治療組,以C57BL/6野生鼠32只作為正常對照組;采用Morris水迷宮進行空間記憶行為學測試;以免疫組化法觀察腦內(nèi)A β 1-42及LRP1的表達,激光共聚焦觀察A β 1-42及LRP1的共表達情況,二次過篩法取微血管段觀察LRP1表達;ELISA法檢測皮層、海馬和血清Aβ1-42,Western Blotting檢測海馬LRP1表達水平,免疫組化和RT-PCR法用來檢測海馬MEOX2、MYOCD、SREBP2 表達。3研究結(jié)果3.1 Morris水迷宮檢測顯示:三個月齡逃避潛伏期電針組相對于模型組都有下降趨勢。重復測量統(tǒng)計顯示:5月齡組間效應方差分析無統(tǒng)計學意義(P0.05),7月齡組間效應方差分析、時間因素及交互作用均有顯著有明顯差異(P0.01);10月齡組間效應方差分析、時間因素、交互作用均有統(tǒng)計學意義(P0.05)三個月齡穿越平臺次數(shù)及平臺象限游泳路程,模型組均明顯低于正常對照組(P0.05),針刺組高于模型組。3.2 7月齡Morris水迷宮檢測顯示:模型組與正常對照組比較逃避潛伏時增加、空間探索實驗穿越平臺次數(shù)、平臺象限游泳路程明顯減少(P0.05,P0.01),電針治療組逃避潛伏時明顯減少(P0.05);激光共聚焦和免疫組化顯示:模型組有明顯的致密性老年斑沉積,針刺組相對減少;電針組LRP1在血管周圍表達相對增多。3.3 ELISA檢測皮層、海馬和血清A β 1-40、Aβ1-42電針治療組較模型組都明顯降低(P0.01)。WB和ELISA結(jié)果顯示LRP1表達水平,模型組低于正常組(P0.01),而電針組高于模型組。3.4PCR結(jié)果顯示MEOX2表達水平,模型組低于正常組(P0.01),而電針組高于模型組(P0.01);MYOCD、SREBP2的表達,模型組高于正常組(P0.01),而電針組低于模型組(P0.01);與免疫組化表達結(jié)果相一致。4結(jié)論4.1電針可以改善APP/PS1雙轉(zhuǎn)基因鼠的學習記憶能力,6月齡可能是電針早期干預AD的最佳治療介入點,但仍需進一步實驗機制研究討論。4.2電針可降低皮層、海馬和血清中A β水平,且電針組具有神經(jīng)毒性的A β 1-42的水平降低較明顯,這可能是電針治療癡呆的作用途徑之一。4.3電針可以改善APP/PS1雙轉(zhuǎn)基因小鼠的學習記憶能力,降低腦內(nèi)Aβ的表達,其機制可能與A β轉(zhuǎn)運受體LRP1水平升高有關。4.4電針上調(diào)MEOX2,降低MYOCD、SREBP2來干預A β轉(zhuǎn)運受體LRP1相關,這可能是電針干預AD血腦屏障清除A β治療AD的可能機制。
[Abstract]:1 Alzheimer's Disease (AD), also known as Alzheimer's disease, is a central nervous system degenerative disease characterized by progressive cognitive impairment and memory impairment. Alzheimer's disease is a major problem facing public health, and has been identified as a key disease in the field of central nervous system injury. Therefore, the study of Alzheimer's disease and related diseases has become an urgent and urgent task, which has become one of the key topics in medical research both at home and abroad. Clinical studies have shown that acupuncture can improve the mental and behavioral state and cognitive function of AD patients. Research and evidence suggest that acupuncture can effectively improve AD, but its mechanism is not clear. It is necessary to further explore the.A beta cascade hypothesis that the formation and clearance of beta amyloid (beta Amyloid protein, A beta) is the cause of its deposition of amyloid plaques in the brain tissue, and its possible neurotoxicity triggering neuronal degeneration. Sex, leading to the emergence of AD symptoms. At present, many teams have studied and proved that they do exist neurotoxicity from multiple angles. A beta plays a very important role in the process of neural degeneration and apoptosis. In this group, the behavior of Electroacupuncture on APP/PS1 transgenic mice of different months of age is studied by Electroacupuncture "Baihui, Yongquan" as the treatment method. To explore the best intervention time of electroacupuncture in treating Alzheimer's disease, the expression of A beta 1-40 and A beta 1-42 in the brain and serum of the APP/PS1 transgenic AD model based on the Electroacupuncture of A0 based on the blood brain barrier clearance pathway, and to explore whether the electroacupuncture can be used to regulate the -1 (LRP1) related genes of the low density lipoprotein receptor protein receptor protein of the APP/PS1 double transgenic mice. To promote the clearance of amyloid protein in the hippocampus and explore the mechanism.2 research method of electroacupuncture treatment of Alzheimer's disease first, 20 4,6,9 month old APP/PS1 double transgenic mice were randomly divided into model group, electroacupuncture treatment group, 10 of the same month old C57BL/6 wild rats as the normal control group; after 6 weeks of Electroacupuncture intervention, the Morris water maze was used. 6 month old APP/PS1 double transgenic mice were randomly divided into model group, electroacupuncture treatment group and 32 C57BL/6 wild rats as normal control group, and Morris water maze was used to conduct spatial memory behavior test, and A beta 1-4 in brain was observed by immunohistochemical method. 2 and LRP1 expression, confocal laser confocal observation of the co expression of A beta 1-42 and LRP1, two times of screening microvascular segment to observe the expression of LRP1, ELISA method to detect the cortex, hippocampus and serum A beta 1-42, Western Blotting to detect the expression level of hippocampus LRP1, immunohistochemistry and RT-PCR method used to test the hippocampus MEOX2, MYOCD, 3.1 expresses the result of MYOCD. The is water maze test showed that the three month old escape incubation group had a downward trend relative to the model group. The repeated measurements showed that there was no significant difference in variance analysis between the 5 month old groups (P0.05), and there were significant differences between the 7 month old groups of effect variance analysis, time factors and interaction (P0.01), and the variance of the effect between the 10 month old groups. The analysis, time and interaction were statistically significant (P0.05) three months of age crossing platform and platform quadrant swimming distance, the model group was significantly lower than the normal control group (P0.05), and the acupuncture group was higher than the model group.3.2 7 month old Morris water maze test. The model group was compared with the normal control group, and the space exploration was higher than the normal control group. The number of traversing platform, the platform quadrant swimming path decreased significantly (P0.05, P0.01), the electroacupuncture treatment group decreased obviously (P0.05). Laser confocal and immunohistochemical staining showed that the model group had obvious densification of the senile plaque and the relative decrease in the acupuncture group; the expression of LRP1 in the electroacupuncture group was relatively increased by.3.3 ELISA to detect the cortex and the sea. The A beta 1-40 and A beta 1-42 electroacupuncture group were significantly lower than the model group (P0.01) and the results of.WB and ELISA showed LRP1 expression level, the model group was lower than the normal group (P0.01), and the electroacupuncture group was higher than the model group, and the.3.4PCR results showed the MEOX2 expression level, the model group was lower than the normal group (P0.01), and the electroacupuncture group was higher than the model group (P0.01); MYOCD, the table of the.3.4PCR group was higher than the model group. The model group was higher than the normal group (P0.01), but the electroacupuncture group was lower than the model group (P0.01), and the.4 conclusion 4.1 electroacupuncture could improve the learning and memory ability of the APP/PS1 double transgenic mice. The 6 month old may be the best intervention point for the early intervention of AD by electroacupuncture, but the further experimental mechanism was still needed to discuss the reduction of.4.2 electroacupuncture. The level of A beta in the lower cortex, hippocampus and serum, and the level of neurotoxic A beta 1-42 in the electroacupuncture group decreased significantly. This may be one of the ways of treating dementia by electroacupuncture..4.3 electroacupuncture can improve the learning and memory ability of APP/PS1 double transgenic mice and reduce the expression of A beta in the brain. The mechanism may be associated with the LRP1 level of A beta transporter LRP1. High related.4.4 electroacupuncture up regulation of MEOX2, reduced MYOCD, and SREBP2 to interfere with LRP1 related A beta transporter receptor, which may be a possible mechanism for the interference of electroacupuncture in the AD blood-brain barrier to eliminate A beta in the treatment of AD.

【學位授予單位】：北京中醫(yī)藥大學
【學位級別】：博士
【學位授予年份】：2017
【分類號】：R245;R-332

【參考文獻】

相關期刊論文 前10條

1 趙鵬;孫亞平;陳紅;董貴成;;阿爾茨海默病發(fā)病機制探究[J];中風與神經(jīng)疾病雜志;2016年01期

2 劉桑;;參芎膠囊治療老年癡呆64例臨床療效觀察[J];中國現(xiàn)代醫(yī)藥雜志;2015年09期

3 吳紅彥;李海龍;顧靜;蘭美華;王虎平;車敏;鄧健男;;黑逍遙散對Aβ_(25~35)誘導AD模型大鼠腦組織神經(jīng)遞質(zhì)及海馬病理改變的影響[J];中國老年學雜志;2015年16期

4 韓向博;王鑫;姜婧;周源;李志剛;;電針對SAMP8小鼠海馬區(qū)PS1蛋白表達的影響[J];現(xiàn)代生物醫(yī)學進展;2015年22期

5 吳羽楠;陳吉祥;陶靜;林如輝;張穎錚;卓沛元;陳立典;;電針百會對APP/PS1轉(zhuǎn)基因小鼠學習記憶能力及Tau蛋白磷酸化的影響[J];中國康復醫(yī)學雜志;2015年05期

6 加吾拉.阿不力孜;王鑫;李芙;白楊;姚海江;莫雨平;周源;許紅;毛穎秋;高譽珊;張偉;張忠;李志剛;薛衛(wèi)國;;觀察電針“百會”、“涌泉”兩穴對不同月齡APP/PS1雙轉(zhuǎn)基因小鼠海馬Aβ淀粉樣沉淀及超微結(jié)構的影響[J];環(huán)球中醫(yī)藥;2015年05期

7 姜婧;高凱;周源;韓向博;樸贊勛;王鑫;曹瑾;盧夢晗;邵千楓;劉剛;李志剛;;Micro-PET與Morris水迷宮觀察“通督啟神”針法對AD模型小鼠療效的影響[J];中華中醫(yī)藥雜志;2015年05期

8 王康鋒;張立娟;陳新勇;;電針大椎及百會穴治療老年性癡呆36例臨床觀察[J];中華中醫(yī)藥雜志;2015年03期

9 李芙;李麗娜;王鑫;白楊;加吾拉·阿不力孜;步青云;高堂珂;薛衛(wèi)國;;電針“百會”“涌泉”對APP/PS 1雙轉(zhuǎn)基因小鼠海馬β淀粉樣蛋白及低密度脂蛋白受體相關蛋白-1水平的影響[J];針刺研究;2015年01期

10 王輝;何婭;陳松盛;馬巧亞;王銳利;;電針對阿爾茨海默病大鼠認知功能及海馬Nogo-A、NgR表達的影響[J];陜西醫(yī)學雜志;2015年02期

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