本文選題：老年性癡呆 + 泛素��； 參考：《湖北中醫(yī)藥大學(xué)》2016年博士論文

【摘要】：目的:補(bǔ)腎化痰開(kāi)竅法是中醫(yī)學(xué)的重要治法之一,在老年性癡呆的防治過(guò)程中起著重要作用。本文系統(tǒng)回顧和總結(jié)了古代醫(yī)家對(duì)老年性癡呆的認(rèn)識(shí),認(rèn)為此病的基本病機(jī)是痰蒙清竅,腎元虧虛,屬本虛標(biāo)實(shí)之證。從腎虛痰蒙清竅理論探討老年性癡呆的發(fā)病機(jī)理和治療法則,本課題以APP/PS1雙轉(zhuǎn)基因小鼠作為老年性癡呆動(dòng)物模型,觀察補(bǔ)腎化痰開(kāi)竅法對(duì)小鼠學(xué)習(xí)記憶水平、腦組織病理形態(tài)和泛素-蛋白酶體通路相的影響,探討補(bǔ)腎化痰開(kāi)竅法對(duì)該模型的干預(yù)作用和作用機(jī)理,旨在為補(bǔ)腎化痰開(kāi)竅法防治老年病的研究提供理論基礎(chǔ)和實(shí)驗(yàn)依據(jù)。方法:1.理論探討:整理與中醫(yī)補(bǔ)腎化痰開(kāi)竅法有關(guān)的古今文獻(xiàn),分析補(bǔ)腎化痰開(kāi)竅法的概念、內(nèi)涵,探討補(bǔ)腎化痰開(kāi)竅法的理論基礎(chǔ),及其防治老年性癡呆的理論依據(jù)。2.以APP/PS1雙轉(zhuǎn)基因小鼠為老年性癡呆動(dòng)物模型,泛素-蛋白酶體途徑為研究靶點(diǎn),研究補(bǔ)腎化痰開(kāi)竅法對(duì)老年性癡呆的作用及其機(jī)制。觀察跳臺(tái)測(cè)試、Morris水迷宮測(cè)試對(duì)小鼠為行為學(xué)影響,檢測(cè)小鼠的學(xué)習(xí)記憶能力;HE染色、免疫組織化學(xué)染色法觀測(cè)小鼠海馬組織泛素-蛋白酶體途徑病理學(xué)變化,用Western Blot法檢測(cè)小鼠海馬組織泛素-蛋白酶體途徑主要蛋白的含量;以Real Time-PCR檢測(cè)小鼠海馬組織泛素m RNA的表達(dá)。結(jié)果:1.Morris水迷宮測(cè)試結(jié)果:空白組比較,模型組小鼠發(fā)現(xiàn)平臺(tái)時(shí)間明顯延長(zhǎng)(P0.01),游泳總路程明顯增加(P0.01);與模型組比較,各組小鼠發(fā)現(xiàn)平臺(tái)時(shí)間均縮短(P0.05),游泳總路程均減少(P0.05);各給藥組與多奈哌齊組比較各組小鼠發(fā)現(xiàn)平臺(tái)時(shí)間未見(jiàn)明顯差異(P0.05),游泳總路程未見(jiàn)明顯差異(P0.05)。益智溫膽組與菖蒲益智丸組比較,兩組小鼠發(fā)現(xiàn)平臺(tái)時(shí)間未見(jiàn)明顯差異(P0.05),游泳總路程未見(jiàn)明顯差異(P0.05)。2.跳臺(tái)測(cè)試實(shí)驗(yàn)結(jié)果給藥90天后,與空白組比較,模型組小鼠的在跳臺(tái)中錯(cuò)誤次數(shù)明顯增加(P0.01),潛伏期明顯縮短(P0.01)。與模型組比較,多奈哌齊組和其它各組小鼠的出錯(cuò)次數(shù)明顯減少(P0.01),多奈哌齊組、益智溫膽組和菖蒲益智丸組小鼠的潛伏期明顯延長(zhǎng)(P0.01)。與多奈哌齊組比較,益智溫膽組和菖蒲益智丸組小鼠的出錯(cuò)次數(shù)和潛伏期無(wú)明顯差別(P0.05);補(bǔ)腎組化痰組出錯(cuò)次數(shù)和潛伏期有所差別(P0.05),化痰組潛伏期有所差別(P0.05)。3.HE染色模型組主要表現(xiàn)為較明顯的腦神經(jīng)元變性、可見(jiàn)到神經(jīng)纖維纏結(jié)和淀粉樣蛋白沉積斑塊。多奈哌齊組和補(bǔ)腎組對(duì)模型轉(zhuǎn)基因小鼠海馬病理形態(tài)影響不明顯,多奈哌齊組僅僅可以觀察到神經(jīng)元數(shù)量變化,以及細(xì)胞結(jié)構(gòu)排列較為整齊,補(bǔ)腎組神經(jīng)元形態(tài)排列較為整齊,結(jié)構(gòu)較多奈哌齊組完整。益智溫膽組、菖蒲益智丸組、化痰組對(duì)神經(jīng)元的保護(hù),空泡變性以及細(xì)胞核增大也有一定好轉(zhuǎn),神經(jīng)細(xì)胞排列以及細(xì)胞形態(tài)明顯較好,神經(jīng)纖維纏結(jié)不明顯。4.免疫組化泛素陽(yáng)性免疫產(chǎn)物定位于海馬區(qū)神經(jīng)元細(xì)胞漿內(nèi)。免疫組化結(jié)果顯示模型組、多奈哌齊組切片內(nèi)海馬CA1區(qū)可見(jiàn)大量泛素,補(bǔ)腎組海馬CA1區(qū)也可見(jiàn)到有部分泛素的聚集;益智溫膽組與菖蒲益智丸組相差不多,二者最少。空白組小鼠海馬CA1區(qū)灰度值明顯較低(P0.01);而多奈哌齊組與其相差無(wú)幾無(wú)明顯差異(P0.05);益智溫膽組、補(bǔ)腎組、化痰組與菖蒲益智丸組小鼠海馬CA1區(qū)灰度值與其有明顯差異(P0.01)。5.小鼠海馬組織Western Blot實(shí)驗(yàn)結(jié)果APP/PS1雙轉(zhuǎn)基因小鼠海馬組織泛素抗體蛋白質(zhì)印跡檢測(cè)結(jié)果表示:與空白組比較,模型組小鼠海馬組織泛素相對(duì)表達(dá)量有顯著性差別(P0.01)。與模型組比較,多奈哌齊組海馬組織泛素相對(duì)表達(dá)量未見(jiàn)顯著性差異(P0.05);模型組較其它各組均顯示表達(dá)量增加。6.小鼠海馬組織Real Time-PCR實(shí)驗(yàn)結(jié)果與空白組比較,模型組泛素m RNA表達(dá)明顯上調(diào),益智溫膽組、補(bǔ)腎組、化痰組、菖蒲益智丸組泛素m RNA均有不同程度的上調(diào)。與模型組比較,多奈哌齊組、益智溫膽組、補(bǔ)腎組、化痰組、菖蒲益智丸組泛素m RNA均有不同程度的下調(diào)。結(jié)論:1.腎虛痰蒙清竅是老年性癡呆的基本病機(jī),補(bǔ)腎化痰開(kāi)竅法能夠有效的防治老年性癡呆。2.補(bǔ)腎化痰開(kāi)竅法對(duì)APP/PS1雙轉(zhuǎn)基因小鼠的學(xué)習(xí)記憶能力具有改善作用。3.補(bǔ)腎化痰開(kāi)竅法能有效的改善動(dòng)物海馬泛素化通路功能,
[Abstract]:Objective: the method of invigorating the kidney and eliminating phlegm and opening the orifices is one of the important treatment methods of traditional Chinese medicine and plays an important role in the prevention and treatment of Alzheimer's disease. This paper systematically reviews and summarizes the understanding of the Alzheimer's disease by ancient doctors. It is believed that the basic pathogenesis of this disease is phlegm, Mongolia, deficiency of the kidney and the deficiency of the kidney. The pathogenesis and treatment rules of Alzheimer's disease were treated with APP/PS1 double transgenic mice as the animal model of Alzheimer's disease. The effect of tonifying kidney and eliminating phlegm on the learning and memory level, the pathological morphology of brain tissue and the effect of ubiquitin proteasome pathway were observed. The effect and effect of the method of invigorating kidney and removing phlegm and opening the orifices on the model were discussed. The mechanism is designed to provide theoretical basis and experimental basis for the study on the prevention and treatment of senile disease by Tonifying the kidney and eliminating phlegm and opening the orifices. Method: 1. theoretical discussion: to organize the ancient and modern literature related to the method of invigorating the kidney and chemical phlegm, analyzing the concept and connotation of the method of invigorating the kidney and removing phlegm and opening the orifices, discussing the theoretical basis of the method of invigorating the kidney and resolving phlegm, and the theory of the prevention and treatment of senile dementia. According to.2., APP/PS1 double transgenic mice were used as the animal model of Alzheimer's disease, and the ubiquitin proteasome pathway was used as the target to study the effect and mechanism of kidney tonifying and phlegm orifices on Alzheimer's disease. The effect of Morris water maze test on behavioral learning in mice was observed, and the learning and memory ability of mice was detected, HE staining and immune tissue were observed. The pathological changes of ubiquitin proteasome pathway in hippocampal tissues of mice were observed by chemical staining. The content of the main proteins of ubiquitin proteasome pathway in hippocampal tissues of mice was detected by Western Blot method; the expression of ubiquitin m RNA in hippocampus of mice was detected by Real Time-PCR. Results: the results of 1.Morris water maze test: blank group comparison, model group mice It was found that the time of the platform was prolonged (P0.01) and the total distance of swimming was significantly increased (P0.01). Compared with the model group, the mice found the platform time shortened (P0.05) and the total swimming course decreased (P0.05). There was no significant difference (P0.05) between the group and the donepezil group (P0), and there was no significant difference in the total swimming course (P0). .05). Compared with the Acorus Yizi group, the two groups of mice found no significant difference (P0.05), and there was no significant difference in the total distance between the two groups (P0.05). After 90 days of the test, the number of errors in the model group was significantly increased (P0.01) and the incubation period was significantly shorter than that in the blank group (P0.01). In the model group, the number of errors in the donepezil group and the other mice decreased significantly (P0.01). The latency of the mice in the donepezil group, the Yizi Wendy group and the calamus Yizi pill group were obviously prolonged (P0.01). There was no significant difference between the Yizi Wendy group and the calamus Yizi pill group (P0.05). There was a difference in the frequency of error and incubation period in the kidney group (P0.05), and the latency of the phlegm group was different (P0.05) in the.3.HE staining model group, the most obvious degeneration of the brain neurons, the neurofibrillary tangles and amyloid plaques, and the effect of donepezil group and kidney supplement on the hippocampus pathological morphology of model transgenic mice was not affected. It was obvious that the number of neurons in the donepezil group could only be observed, and the structure of the cells was neatly arranged. The structure of the tonifying group was more neat and the structure was more complete. The protection of the neurons, the vacuoles degeneration and the nucleus enlargement of the group of Yizi wendyen, the calamus Yizi pill group and the phlegm group were also improved. The cell morphology and cell morphology were better, and the neurofibrillary tangles were not obvious in the hippocampal neuronal cytoplasm. The immunohistochemical results showed that in the model group, a large number of ubiquitin was found in the hippocampal CA1 area in the donepezil section, and some ubiquitin aggregation was found in the hippocampus CA1 area of the tonifying kidney group. The difference between the Yizi Wendy group and the calamus Yizi pill group was not much, the two was the least. The gray value of the hippocampal CA1 area in the blank group was significantly lower (P0.01), but there was no significant difference between the donepezil group and the group of donepezil (P0.05); the gray value of the hippocampus CA1 area of the rats of the Yizi Wendan group, the kidney tonifying group, the phlegm group and the calamus Yizi pill group was significantly different from that of the.5. mice (P0.01). Western Blot test results of hippocampal tissue in the hippocampus of APP/PS1 double transgenic mice showed that: compared with the blank group, there was a significant difference in the relative expression of ubiquitin in the hippocampus of the model group (P0.01). Compared with the model group, the relative expression of ubiquitin in the hippocampus of donepezil group was not significant difference. Compared with the other groups, the model group showed an increase in the expression of Real Time-PCR in the hippocampus of.6. mice compared with the blank group. The expression of ubiquitin m RNA in the model group was obviously up-regulated, and the level of ubiquitin m RNA in the Yizi Wendan group, the tonifying group, the phlegm group and the Acorus Yizi pill group were up to be up. Compared with the model group, the donepezil group and the beneficial intelligence were improved. The ubiquitin m RNA in the group of warm gallbladder, kidney tonifying, phlegm and Yizi Yizhi group have different downgrades. Conclusion: 1. the kidney deficiency and phlegm Mongolia clear orifices are the basic pathogenesis of senile dementia, the method of invigorating the kidney and eliminating phlegm and opening the orifices can effectively prevent and control the.2. invigorating phlegm and the orifices of the senile dementia, which can improve the learning and memory ability of the APP/PS1 double transfer gene mice,.3. Tonifying kidney, resolving phlegm and inducing resuscitation can effectively improve the function of hippocampus ubiquitination pathway in animals.
【學(xué)位授予單位】：湖北中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類(lèi)號(hào)】：R277.7

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2 王玉娜;補(bǔ)腎化痰活血法治療多囊卵巢綜合征的文獻(xiàn)研究[D];黑龍江中醫(yī)藥大學(xué);2010年

3 楊雪;補(bǔ)腎化痰通經(jīng)湯治療腎虛痰瘀型青春期閉經(jīng)臨床觀察[D];湖北中醫(yī)藥大學(xué);2012年

4 王琳琳;補(bǔ)腎化痰通絡(luò)方延緩原發(fā)性高血壓左室重構(gòu)的療效觀察[D];山東中醫(yī)藥大學(xué);2014年

5 楊蘭;針刺結(jié)合補(bǔ)腎化痰方治療多囊卵巢綜合征的臨床療效觀察[D];南京中醫(yī)藥大學(xué);2013年

6 李文英;“補(bǔ)腎化痰益氣活血”法預(yù)防腦缺血神經(jīng)保護(hù)作用的實(shí)驗(yàn)研究[D];黑龍江中醫(yī)藥大學(xué);2003年

7 戚敏;補(bǔ)腎化痰序貫療法對(duì)PCOS患者血清Apelin表達(dá)的影響[D];山東中醫(yī)藥大學(xué);2014年

8 賀亞蕾;補(bǔ)腎化痰方對(duì)PCOS大鼠糖脂代謝功能的影響[D];河南中醫(yī)學(xué)院;2014年

9 鄭長(zhǎng)春;補(bǔ)腎化痰法聯(lián)合二甲雙胍治療腎虛痰阻型PCOS伴IR臨床觀察[D];湖北中醫(yī)藥大學(xué);2012年

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