本文選題：β淀粉樣蛋白 + 阿爾茨海默病　； 參考：《遼寧中醫(yī)藥大學(xué)》2015年博士論文

【摘要】：目的:通過高脂飲食建立高脂致Aβ增高小型豬動物模型,觀察高膽固醇血癥膽固醇代謝異常狀態(tài)對腦內(nèi)β淀粉樣蛋白生成的影響,探討膽固醇代謝異常狀態(tài)下腦膽固醇代謝紊亂相關(guān)因素與AD早期病理改變β淀粉樣蛋白生成的關(guān)系;運用具有不同功效的中藥復(fù)方干預(yù)此動物模型,并對其作用機制進(jìn)行探討,為在早期階段通過減少AD典型病理特征Aβ生成及沉積從而防治AD提供實驗依據(jù)。材料與方法:廣西巴馬小型豬55只,月齡6-8個月,體重(29.88±2.27)kg,雄性,腸道菌群正常,體健,由泰州泰和生物科技有限公司提供。倫理符合中華人民共和國衛(wèi)生部動物實驗管理條例(No.55,2001)和遼寧中醫(yī)藥大學(xué)實驗動物管理條例規(guī)定。實驗室溫度為(20.80±2.97)℃,濕度(47.30±15.31)%,實驗動物均為單籠飼養(yǎng)。適應(yīng)性飼養(yǎng)1周后,采用隨機數(shù)字表法將55只小型豬分為兩組:正常組5只,手術(shù)組50只。正常組給予基礎(chǔ)飲食,手術(shù)組(第2周進(jìn)行冠脈球囊擠壓伴拉傷,術(shù)前禁食8h,禁水2小時),術(shù)后繼續(xù)予以高熱量高脂適量膽鹽喂飼(防止小豬連續(xù)吃大量油膩食物出現(xiàn)厭食,加膽鹽是防止消化不良)。日進(jìn)食量為上周所稱體重的3%,自由飲水。術(shù)中有8只小型豬死亡;應(yīng)用隨機數(shù)字表將存活的42只小型豬重新進(jìn)行隨機分組:模型組(n=7)、健脾祛痰化瘀組(n=7)、健脾祛痰組(n=7)、健脾組(n=7)、祛痰化瘀組(n=7)、西藥組(n=7)至此,健脾組加入健脾方,健脾祛痰組加入健脾祛痰方,祛痰化瘀組加入祛痰化瘀方,健脾祛痰化瘀復(fù)方組加入健脾祛痰化瘀方,加入相應(yīng)劑量的中藥復(fù)方按照體重對應(yīng)標(biāo)準(zhǔn),飼喂至24周,指標(biāo)檢測如下:1.實驗期間每4周稱量小型豬的體重,觀察小型豬體重變化情況。2.實驗第0周及實驗開始后第4周,第12周,第24周禁食16h后耳緣靜脈抽血3ml,全自動生化分析儀檢測血脂(TC、TG、LDL、HDL)水平,觀察不同時間點血脂四項的變化情況。3.免疫組織化學(xué)染色法檢測Aβ陽性細(xì)胞在大腦組織的分布。4.免疫組織化學(xué)法觀察Aβ在大腦組織的含量及水平。5.免疫組織化學(xué)法檢測Aβ代謝相關(guān)因子BACE1及腦膽固醇代謝關(guān)鍵因子HMG-Co AR、CYP46a、ABCA1在腦內(nèi)的含量及在神經(jīng)元細(xì)胞內(nèi)的表達(dá)。6.免疫蛋白印跡法(Western Blot)檢測Aβ代謝相關(guān)因子BACE1及腦膽固醇代謝關(guān)鍵因子HMG-Co AR、CYP46a、Apo E、ABCA1的蛋白表達(dá)。本實驗所有數(shù)據(jù)結(jié)果使用SPSS17.0軟件進(jìn)行統(tǒng)計分析,結(jié)果用平均值加減標(biāo)準(zhǔn)差(—X±S)表示,組與組之間比較采用單因素方差分析。(P0.05說明有顯著性差異,P0.01說明有非常顯著性差異。)結(jié)果:1.正常組與模型組小型豬的體重增長變化顯示:模型組與正常組在第0周體重增長并無明顯差異,兩組巴馬小型豬體重增長從第4周起開始有顯著性差異(P0.05);2.正常小豬在成長過程中,血脂的變化不明顯;而模型組巴馬小型豬血脂四項變化顯著,結(jié)果顯示:高脂飲食1周,可使小型豬甘油三酯升高2.86倍,使總膽固醇升高3.84倍,使低密度脂蛋白升高7.40倍,使高密度脂蛋白升高2.56倍;隨著高脂飲食時間延長,小型豬甘油三酯(TG)、總膽固醇(TC)、低密度脂蛋白(LDL)持續(xù)升高,第24周趨于穩(wěn)定,而高密度脂蛋白隨高脂飲食時間增加呈降低趨勢;3.免疫組織化學(xué)法顯示高脂飲食造成的高血膽固醇水平增加了腦Aβ含量,模型組巴馬小型豬腦Aβ含量顯著高于正常組(0.26±0.03 pg/mg protein vs 0.1110±0.01 pg/mg protein,p0.01)。免疫組織化學(xué)法檢測平均光密度值(AOD)正常組與模型組相比較,有統(tǒng)計學(xué)意義。(P0.01)。免疫組織化學(xué)染色顯示與正常組相比較,模型組腦組織神經(jīng)元細(xì)胞結(jié)構(gòu)呈棕黃色顆粒聚集,表示腦組織內(nèi)Aβ陽性細(xì)胞表達(dá)增多;4.健脾祛痰組調(diào)節(jié)TC、HDL、LDL的能力強于其余兩組,健脾祛痰化瘀組調(diào)節(jié)TG的能力強于其余兩組;5.與模型組比較,健脾組、健脾祛痰組、祛痰化瘀組、健脾祛痰化瘀組均能明顯降低腦內(nèi)Aβ含量(P0.01);健脾祛痰化瘀組較健脾組、健脾祛痰組、祛痰化瘀組減少Aβ含量,且有統(tǒng)計學(xué)意義(P0.01)健脾組、健脾祛痰組、祛痰化瘀組之間比較,無統(tǒng)計學(xué)差異(P0.05);6.經(jīng)健脾祛痰化瘀法干預(yù)后,健脾組、健脾祛痰組、祛痰化瘀組、健脾祛痰化瘀組腦內(nèi)BACE1蛋白表達(dá)明顯降低;證明血膽固醇降低引起的腦Aβ降低與Aβ生成相關(guān)因子的蛋白表達(dá)的改變有關(guān);7.免疫組織化學(xué)法檢測ABCA1實驗結(jié)果顯示,與正常組相比較,模型組及中藥復(fù)方各組腦組織ABCA1含量增加,差異有統(tǒng)計學(xué)意義(P0.01);與模型組相比較,健脾組、健脾祛痰組、祛痰化瘀組、健脾祛痰化瘀組ABCA1含量升高,差異有統(tǒng)計學(xué)意義(P0.01);與健脾祛痰化瘀組相比,健脾組、健脾祛痰組、祛痰化瘀組ABCA1含量降低,差異有統(tǒng)計學(xué)意義(P0.01);健脾組、健脾祛痰組、祛痰化瘀組組間比較差異無統(tǒng)計學(xué)意義(P0.01)。免疫組化染色顯示模型組陽性細(xì)胞表達(dá)增多,而藥物組陽性細(xì)胞表達(dá)減少,與模型組相比,差異有統(tǒng)計學(xué)意義(P0.01);8.免疫組織化學(xué)法檢測HMG-Co AR結(jié)果顯示,與正常組相比,模型組及中藥復(fù)方各組腦組織HMG-Co A含量增加,且有顯著性差異(P0.01);與模型組相比較,健脾組、健脾祛痰組、祛痰化瘀組及健脾祛痰化瘀組HMG-Co AR含量增加,差異有統(tǒng)計學(xué)意義(P0.01);與健脾祛痰化瘀組相比,健脾組,健脾祛痰組、祛痰化瘀組HMG-Co AR含量減少,差異有統(tǒng)計學(xué)意義(P0.01);健脾組,健脾祛痰組,祛痰化瘀組之間比較,HMG-Co AR差異無統(tǒng)計學(xué)意義(P0.01),免疫組化染色顯示模型組陽性細(xì)胞表達(dá)增多,而藥物組陽性細(xì)胞表達(dá)減少,與模型組相比,差異有統(tǒng)計學(xué)意義;9.免疫組織化學(xué)法檢測CYP46a結(jié)果顯示,與正常組相比較,模型組、復(fù)方各組腦組織CYP46a含量增加,且有顯著性差異(P0.01);與模型組相比較,復(fù)方組腦組織內(nèi)CYP46a含量增加,差異有統(tǒng)計學(xué)意義(P0.01);與健脾祛痰化瘀組相比,健脾組、健脾祛痰組、祛痰化瘀組腦組織CYP46a含量減少,差異有統(tǒng)計學(xué)意義(P0.01);與健脾組相比,健脾祛痰組、祛痰化瘀組腦組織CYP46a含量增加,且差異有統(tǒng)計學(xué)意義(P0.01);與健脾祛痰組相比,祛痰化瘀組CYP46減少,差異無統(tǒng)計學(xué)意義。(P0.05)。免疫組化染色顯示模型組陽性細(xì)胞表達(dá)增多,而藥物組陽性細(xì)胞表達(dá)減少,與模型組相比,差異有統(tǒng)計學(xué)意義(P0.01);10.Western Blot結(jié)果顯示,與正常組相比較,模型組、中藥復(fù)方組腦組織HMG-Co AR蛋白表達(dá)升高,且差異有統(tǒng)計學(xué)意義(P0.01);與模型組相比較,復(fù)方組HMGR蛋白表達(dá)升高,且差異有統(tǒng)計學(xué)意義(P0.01);與健脾祛痰化瘀組相比,健脾組、健脾祛痰組、祛痰化瘀組腦組織HMG-Co AR蛋白表達(dá)降低,差異有統(tǒng)計學(xué)意義(P0.01)與健脾組相比,健脾祛痰組、祛痰化瘀組HMG-Co AR蛋白表達(dá)升高,差異有統(tǒng)計學(xué)意義(P0.05);健脾祛痰組、祛痰化瘀組相比,HMGR蛋白表達(dá)差異無統(tǒng)計學(xué)意義(P0.05);11.Western Blot結(jié)果顯示,與正常組相比,模型組、復(fù)方組腦組織CYP46a蛋白表達(dá)升高,差異有統(tǒng)計學(xué)意義(P0.01);與模型組相比較,復(fù)方組腦組織內(nèi)CYP46a1蛋白表達(dá)均有升高,差異有統(tǒng)計學(xué)意義(P0.01);與健脾祛痰化瘀相比,祛痰化瘀組、健脾祛痰組及健脾組腦組織CYP46a蛋白表達(dá)降低,差異有統(tǒng)計學(xué)意義;健脾組、健脾祛痰組、祛痰化瘀組腦組織CYP46a1蛋白表達(dá)無顯著性差異。(P0.05);12.Western Blot結(jié)果顯示,與正常組相比較,模型組、復(fù)方組腦組織Apo E蛋白表達(dá)升高,且有顯著性差異(P0.01);與模型組相比,復(fù)方組腦組織內(nèi)Apo E蛋白表達(dá)降低,差異有統(tǒng)計學(xué)意義(P0.01);與健脾祛痰化瘀組相比較,祛痰化瘀組、健脾祛痰組及健脾組腦組織Apo E蛋白表達(dá)升高,有顯著性差異(P0.01);健脾組、健脾祛痰組、祛痰化瘀組腦組織蛋白表達(dá)差異無統(tǒng)計學(xué)意義。(P0.05);13.Western Blot結(jié)果顯示,與正常組相比,模型組、復(fù)方組腦組織ABCA1蛋白表達(dá)升高,且有顯著性差異(P0.01);與模型組相比較,復(fù)方組對腦組織內(nèi)ABCA1蛋白表達(dá)升高,差異有統(tǒng)計學(xué)意義(P0.01);與健脾祛痰化瘀組相比,健脾祛痰組、祛痰化瘀組及健脾組腦組織ABCA1蛋白表達(dá)降低,差異有統(tǒng)計學(xué)意義(P0.01);健脾組、健脾祛痰組、祛痰化瘀組之間相比,腦組織ABCA1蛋白表達(dá)無顯著性差異。(P0.05)。結(jié)論:1.高脂飲食可影響血脂四項水平,高脂飲食1周可造成冠狀動脈粥樣硬化模型豬血脂異常,隨高脂飲食時間延長,血脂四項中甘油三酯(TG)、總膽固醇(TC)、低密度脂蛋白(LDL)的濃度升高,高密度脂蛋白(HDL)濃度降低;2.高脂飲食增加了外周血血脂濃度,外周血脂水平可影響巴馬小型豬腦內(nèi)Aβ的水平的改變,使腦內(nèi)Aβ含量增加,高脂飲食24周可建立高脂致Aβ增高動物模型;3.健脾方、健脾祛痰方、祛痰化瘀方、健脾祛痰化瘀方可調(diào)節(jié)巴馬小型豬模型的血脂水平,健脾祛痰化瘀方優(yōu)于健脾、健脾祛痰及祛痰化瘀方;4.健脾方、健脾祛痰方、祛痰化瘀方、健脾祛痰化瘀方可減少Aβ在腦內(nèi)的沉積,減少Aβ的表達(dá);5.健脾方、健脾祛痰方、祛痰化瘀方、健脾祛痰化瘀方減少Aβ含量和生成的可能機制之一是通過降低Aβ代謝關(guān)鍵因子BACE1蛋白表達(dá)來實現(xiàn)的;6.腦膽固醇代謝異常能夠影響Aβ生成與代謝;7.健脾方、健脾祛痰方、祛痰化瘀方、健脾祛痰化瘀方可使腦中膽固醇合成及轉(zhuǎn)出的關(guān)鍵酶Apo E蛋白下調(diào),使HMGR、ABCA1、和CYP46的蛋白表達(dá)上調(diào),其中健脾祛痰化瘀方作用優(yōu)于健脾、健脾祛痰、祛痰化瘀、健脾祛痰化瘀方。8.健脾方、健脾祛痰方、祛痰化瘀方、健脾祛痰化瘀方干預(yù)高脂所致Aβ增高動物模型的可能機制是通過下調(diào)Apo E蛋白表達(dá),上調(diào)HMGR、ABCA1、和CYP46的蛋白表達(dá)來發(fā)揮作用。9.健脾方、健脾祛痰方、祛痰化瘀方、健脾祛痰化瘀方可能是通過調(diào)節(jié)巴馬小型豬的外周血脂水平、調(diào)節(jié)Aβ生成相關(guān)分泌酶β分泌酶的活性及BACE1蛋白表達(dá)、下調(diào)Apo E蛋白表達(dá),上調(diào)CYP46a HMG-Co AR、ABCA1等蛋白的表達(dá)來干預(yù)阿爾茨海默病早期β淀粉樣蛋白形成的病理過程,從而在早期阻抑或延緩AD的發(fā)生發(fā)展。10.健脾方、健脾祛痰方、祛痰化瘀方、健脾祛痰化瘀方在調(diào)脂、調(diào)節(jié)Aβ生成及改善腦膽固醇代謝方面效果顯著,健脾祛痰化瘀方作用大于健脾祛痰方、祛痰化瘀方大于健脾方。說明健脾祛痰化瘀方配伍精良,健脾、祛瘀、化痰三者均不可少。
[Abstract]:Objective: to establish an animal model of high fat induced A beta hyperlipidemia pig by high fat diet, to observe the effect of abnormal cholesterol metabolism in hypercholesterolemia on the formation of beta amyloid in the brain, and to explore the relationship between the related factors of cholesterol metabolism in the abnormal state of cholesterol metabolism and the early pathological changes of beta amyloid protein in AD. In order to provide experimental basis for the prevention and treatment of AD by reducing the typical pathological features of AD, A beta generation and deposition in the early stage, 55 Chinese Bama miniature pigs, 6-8 months of age, weight (29.88 + 2.27) kg, male and normal intestinal flora, were provided for the prevention and treatment of AD in the early stage by reducing the typical pathological characteristics of AD. Body health, provided by Taizhou Taihe biological science and Technology Co., Ltd., the ethics conforms to the regulations of the animal experiment management regulation (No.552001) of the Ministry of health of People's Republic of China and the regulations of the experimental animal management of Liaoning University of Traditional Chinese Medicine. The laboratory temperature is (20.80 + 2.97) and humidity (47.30 + 15.31)%, the experimental animals are kept in single cage. After 1 weeks of adaptive breeding, 55 small pigs were divided into two groups by random digital table: 5 in the normal group and 50 in the operation group. The normal group was given the basic diet, the operation group (second weeks of coronary balloon extrusion accompanied by pull, the preoperative fasting 8h, the water prohibition 2 hours), and the postoperative continuous high calorie, high fat and moderate salt feeding (preventing the piglets to eat a large amount of greasy food in an anorexia. Bile salt is to prevent dyspepsia). Intake of daily intake is 3% of weight of last week's weight, free drinking water. 8 small pigs died in the operation; 42 small pigs survived were randomly divided into model group (n=7), spleen eliminating phlegm and stasis group (n=7), spleen eliminating phlegm group (n=7), spleen invigorating group (n=7), phlegm removing stasis group (n=7), western medicine group (n=7) The Jianpi group joined Jianpi prescription, Jianpi dispelling phlegm group joined Jianpi dispelling phlegm prescription, dispelling phlegm and removing stasis group joined the prescription of eliminating phlegm and removing stasis, the compound group of Jianpi dispelling phlegm and removing stasis was added to Jianpi dispelling phlegm and removing stasis prescription, and the corresponding dosage of Chinese medicine compound was added to the body weight corresponding standard for 24 weeks, and the indexes were measured as follows: the weight of small pigs was weighed every 4 weeks during the 1. experiment. The body weight change of small pigs was examined by.2. zeroth weeks and fourth weeks, Twelfth weeks and twenty-fourth weeks after the experiment. 3ml was extracted after 16h, and the level of blood lipid (TC, TG, LDL, HDL) was detected by automatic biochemical analyzer, and the changes of blood lipid four items at different time points were observed..3. free histochemical staining was used to detect the expression of A beta positive cells in the brain tissue. The content and level of A beta in brain tissue were observed by distributed.4. immunohistochemical method and.5. immunohistochemical staining was used to detect A beta metabolism related factor BACE1 and the key factor of brain cholesterol metabolism HMG-Co AR, CYP46a, ABCA1 in the brain and the expression of.6. immunoblotting (Western Blot) in the neuron cells. BACE1 and the key factors of brain cholesterol metabolism HMG-Co AR, CYP46a, Apo E, ABCA1 protein expression. All data results in this experiment were statistically analyzed using SPSS17.0 software, and the results were added and subtracting the standard deviation (- X + S) with the average value (- X + S), and the group and the group were compared with the single factor variance analysis. (P0.05 shows a significant difference, P0.01 instructions have a very significant difference. Results: the body weight growth of 1. normal groups and model group small pigs showed that there was no significant difference between the model group and the normal group at zeroth weeks. The weight growth of the two groups of Bama miniature pigs began to have significant difference from fourth weeks (P0.05); 2. normal piglets had no significant changes in blood lipid during the growth process; The four changes of blood lipid were significant. The results showed that high fat diet for 1 weeks could increase triglyceride of small pigs by 2.86 times, increase total cholesterol by 3.84 times, increase low density lipoprotein by 7.40 times, make high density lipoprotein (HDL) increase 2.56 times. With the prolongation of high fat diet, small pig triglyceride (TG), total cholesterol (TC), low density lipoprotein (LDL) (LDL) continued to rise, and the twenty-fourth weeks tended to stabilize, while high density lipoprotein decreased with the increase of high fat diet time; 3. the high blood cholesterol level caused by high fat diet increased the content of brain A beta in the high fat diet, and the A beta content in the brain of the model group was significantly higher than that of the normal group (0.26 + 0.03 pg/mg protein vs 0.1110 + 0.01 pg/m G protein, P0.01). Compared with the model group, the average light density (AOD) in the normal group was compared with the model group. (P0.01). (P0.01). Immunohistochemical staining showed that the structure of neurons in the brain tissue of the model group was brown yellow particles gathering, indicating that the expression of A beta positive cells in the brain tissue increased; 4. The ability of regulating TC, HDL, LDL in the phlegm group is stronger than the other two groups, and the ability of regulating TG in the invigorating spleen and removing phlegm and removing stasis group is stronger than the other two groups. 5. compared with the model group, the group of invigorating the spleen, invigorating the spleen and removing phlegm, removing phlegm and removing stasis, and eliminating phlegm and removing stasis group can obviously reduce the content of A beta in the brain (P0.01); the group of Jianpi dispelling phlegm and removing stasis is more than the invigorating spleen group, invigorating the spleen and expectorant group and removing phlegm and removing stasis group. Reduce the content of A beta, and have statistical significance (P0.01) in the invigorating spleen group, invigorating the spleen and eliminating phlegm group, eliminating phlegm and removing stasis group, there is no statistical difference (P0.05); 6. after strengthening the spleen and removing phlegm and removing stasis method, the expression of BACE1 protein in the brain of Jianpi group, Jianpi dispelling phlegm group, dispelling phlegm and removing stasis in the brain of Jianpi dispelling phlegm and removing stasis is obviously reduced, and the brain A beta caused by the decrease of blood cholesterol is proved to be the brain A beta The changes in the protein expression of A beta generation related factors were reduced; 7. the results of ABCA1 test by immunohistochemistry showed that the content of ABCA1 in the brain tissue of the model group and the traditional Chinese medicine compound was increased compared with the normal group, and the difference was statistically significant (P0.01); compared with the model group, the spleen strengthening group, the invigorating spleen and eliminating phlegm group, eliminating phlegm and removing stasis group and invigorating the spleen and removing the spleen were compared with the model group. The content of ABCA1 in the phlegm and removing stasis group was higher, the difference was statistically significant (P0.01). Compared with the invigorating spleen and eliminating phlegm and removing stasis group, the content of ABCA1 in the Jianpi group, the invigorating spleen and eliminating phlegm group and the dispelling phlegm and removing stasis group were statistically significant (P0.01), and there was no significant difference between the Jianpi group, the invigorating spleen and eliminating phlegm group and the expectorant and removing stasis group (P0.01). Immunohistochemical staining display model was not significant. The expression of positive cells in the group increased and the expression of positive cells in the drug group decreased. Compared with the model group, the difference was statistically significant (P0.01). The results of HMG-Co AR by 8. immunohistochemical staining showed that the content of HMG-Co A in the brain tissues of the model group and the traditional Chinese medicine group was significantly higher than that in the normal group, and there were significant differences (P0.01), compared with the model group, The content of HMG-Co AR in Jianpi group, invigorating spleen and eliminating phlegm group, eliminating phlegm and removing stasis group and promoting phlegm and removing stasis group increased with statistical significance (P0.01). Compared with the group of Jianpi dispelling phlegm and removing stasis, the content of HMG-Co AR in Jianpi group, Jianpi dispelling phlegm group and dispelling phlegm and removing stasis group was statistically significant (P0.01), and the comparison between Jianpi group, Jianpi dispelling phlegm group and eliminating phlegm and removing stasis group was compared. The difference of HMG-Co AR was not statistically significant (P0.01). Immunohistochemical staining showed that the expression of positive cells in the model group increased, but the expression of positive cells in the drug group decreased, and the difference was statistically significant compared with the model group. 9. the results of CYP46a detection by immunohistochemistry showed that the content of CYP46a in the brain tissue of the model group and the compound group was increased compared with the normal group. And there was significant difference (P0.01). Compared with the model group, the content of CYP46a in the brain tissue of the compound group increased, and the difference was statistically significant (P0.01). Compared with the invigorating spleen and eliminating phlegm and removing stasis group, the content of CYP46a in the brain tissue of the invigorating spleen group, the invigorating spleen and eliminating phlegm group and the dispelling phlegm and removing stasis group was statistically significant (P0.01); compared with the Jianpi group, the spleen eliminating phlegm group and expectorant were compared with the invigorating spleen group. The content of CYP46a in the brain tissue of the Huayu group increased and the difference was statistically significant (P0.01). Compared with the invigorating spleen and eliminating phlegm group, the difference was not statistically significant (P0.05). (P0.05). Immunohistochemical staining showed that the expression of positive cells in the model group increased and the positive cell expression of the drug group decreased, compared with the model group, the difference was statistically significant (P0. 01); the results of 10.Western Blot showed that the expression of HMG-Co AR protein in the brain tissue of the model group and the compound group was higher than that of the normal group, and the difference was statistically significant (P0.01). Compared with the model group, the expression of HMGR protein in the compound group was higher, and the difference was statistically significant (P0.01). Compared with the group of invigorating the spleen and removing phlegm and removing stasis, the spleen and the spleen and eliminating phlegm group were compared. The expression of HMG-Co AR protein in the brain tissue of the dispelling phlegm and removing stasis group decreased, and the difference was statistically significant (P0.01), compared with the Jianpi group, the expression of HMG-Co AR protein in the group of invigorating the spleen and removing phlegm and removing stasis was statistically significant (P0.05). There was no significant difference in the expression of HMGR protein in the group of invigorating the spleen and eliminating phlegm and removing phlegm and removing stasis (P0.05) and 11.Western Blot junction. Compared with the normal group, the expression of CYP46a protein in the brain tissue of the model group and the compound group increased, and the difference was statistically significant (P0.01). Compared with the model group, the expression of CYP46a1 protein in the brain tissue of the compound group increased, and the difference was statistically significant (P0.01). Compared with the invigorating spleen and eliminating phlegm and removing stasis, the group of dispelling phlegm and removing stasis, strengthening the spleen and eliminating phlegm and invigorating the spleen group brain group The expression of CYP46a protein decreased, and the difference was statistically significant. There was no significant difference in the expression of CYP46a1 protein in the brain tissue of the Jianpi group, the invigorating spleen and eliminating phlegm group and the removing phlegm and removing stasis group. (P0.05); the results of 12.Western Blot showed that the expression of egg white in the model group and the compound group brain tissue was higher than that of the normal group, and there was a significant difference (P0.01) in the model group (P0.01). Compared with the model group, the expression of the protein expression was significantly higher than that of the normal group (P0.01). The expression of Apo E protein in the brain tissue of the compound group decreased (P0.01), and the expression of Apo E protein in the brain tissue of the anti phlegm and removing stasis group, the invigorating spleen and eliminating phlegm group and the invigorating spleen group increased significantly (P0.01), and there was no significant difference in the protein expression of the brain tissue in the Jianpi group, the invigorating spleen and eliminating phlegm group and the dispelling phlegm and removing stasis group. (P0.05); the results of 13.Western Blot showed that the expression of ABCA1 protein in the brain tissue of the model group and the compound group was higher than that in the normal group, and there was a significant difference (P0.01). Compared with the model group, the expression of ABCA1 protein in the brain tissue was higher, the difference was statistically significant (P0.01). Compared with the group of invigorating the spleen and removing phlegm and removing stasis, the group of invigorating the spleen and removing phlegm and removing phlegm and removing stasis The expression of ABCA1 protein in the brain tissue of the group and the invigorating spleen group decreased, and the difference was statistically significant (P0.01). There was no significant difference in the expression of ABCA1 protein in the brain tissue between the Jianpi group, the invigorating spleen and eliminating phlegm group and the dispelling phlegm and removing stasis group. (P0.05) 1. high fat diet could affect the level of blood lipid, and the high fat diet could cause coronary atherosclerosis model pig blood for 1 weeks. Lipid abnormality, with the prolongation of high fat diet time, the concentration of triglyceride (TG), total cholesterol (TC), low density lipoprotein (LDL) and high density lipoprotein (HDL) decreased in four lipids, and 2. high fat diet increased the concentration of blood lipid in peripheral blood, and the level of peripheral blood lipid could affect the level of A beta in the brain of Bama small pig, and increase the content of A beta in the brain. Plus, high fat diet for 24 weeks can establish high fat induced A beta animal model; 3. Jianpi Recipe, invigorating the spleen and eliminating phlegm prescription, dispelling phlegm and removing stasis prescription, invigorating the spleen and Removing Blood Stasis Recipe can regulate the blood lipid level of Bama miniature pig model, strengthening spleen and removing phlegm and removing stasis prescription is superior to the spleen, invigorating the spleen and eliminating phlegm and removing phlegm and removing stasis; 4. invigorating spleen, eliminating phlegm and removing phlegm, eliminating phlegm and removing stasis, invigorating the spleen and removing phlegm and removing stasis It can reduce the deposition of A beta in the brain and reduce the expression of A beta; 5. the prescription of Jianpi Fang, Jianpi Dispelling Phlegm Recipe, dispelling phlegm and removing stasis, reducing the content and formation of A beta by eliminating phlegm and removing stasis is realized by reducing the expression of BACE1 protein, the key factor of A beta metabolism; the 6. brain cholesterol metabolism abnormality can affect the formation and metabolism of A beta; 7. spleen invigorating prescription, The recipe for invigorating the spleen and removing phlegm, eliminating phlegm and removing stasis, can reduce the protein expression of the key enzyme of cholesterol synthesis and transformation in the brain, and make the expression of the protein of HMGR, ABCA1, and CYP46 up regulation. The effect of strengthening spleen and removing phlegm and removing stasis is superior to the spleen strengthening, invigorating the spleen and eliminating phlegm, removing phlegm and removing stasis, strengthening spleen and removing phlegm and removing blood stasis prescription, invigorating spleen and eliminating phlegm, dispelling phlegm and removing stasis The possible mechanism of invigorating spleen and removing phlegm and removing stasis to interfere with high fat A beta animal model is to regulate the expression of Apo E protein, increase the expression of HMGR, ABCA1, and CYP46 to play the role of.9. Jianpi Recipe, invigorate the spleen and dispelling phlegm prescription, dispelling phlegm and removing stasis prescription, the prescription for regulating spleen and removing phlegm and removing stasis may be regulated by the peripheral blood lipid level of Bama miniature pig, regulating A Beta production of secretory enzyme beta secretase activity and BACE1 protein expression, downregulation of Apo E protein expression, up regulation of CYP46a HMG-Co AR, ABCA1 and other proteins
【學(xué)位授予單位】：遼寧中醫(yī)藥大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：R-332;R543.3

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