發(fā)布時(shí)間：2018-04-11 02:40

  本文選題：Aβ + APP代謝酶　； 參考：《天津醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:觀察β片層阻斷肽H102對(duì)APP/PS1雙轉(zhuǎn)基因AD小鼠腦內(nèi)Aβ、APP代謝分泌酶及AD小鼠學(xué)習(xí)記憶能力的影響。方法:1.隨機(jī)將30只6個(gè)月齡的APP/PS1雙轉(zhuǎn)基因小鼠分為AD組和H102組,同月齡和同背景的15只C57BL/6J小鼠作為對(duì)照組。采用鼻腔給藥的方法,同一時(shí)間段,每天給藥一次,H102組小鼠給予H102溶液(5.8 mg/kg)5μl,AD組和對(duì)照組給予輔料溶液5μl;連續(xù)給藥30天后通過(guò)新物體識(shí)別實(shí)驗(yàn)和Morris水迷宮實(shí)驗(yàn)測(cè)試各組小鼠的學(xué)習(xí)記憶能力。2.行為學(xué)實(shí)驗(yàn)完成之后,10%的水合氯醛腹腔注射麻醉,PBS灌流、處死小鼠、冰上取腦,將一側(cè)腦組織放于提前準(zhǔn)備好的4%的多聚甲醛中,24h后進(jìn)行包埋,將包埋好的組織制作成切片,用于免疫組織化學(xué)法檢測(cè)腦內(nèi)ADAM10、ADAM17、BACE1、PS1、PEN-2、APH1-a、Aβ蛋白的表達(dá);另一半將海馬和皮質(zhì)分離后,做好標(biāo)記先短暫保存在液氮中,然后再?gòu)囊旱腥〕龇湃?80℃冰箱保存,用于ELISA檢測(cè)Aβ蛋白的表達(dá)和Western blot檢測(cè)腦內(nèi)ADAM10、ADAM17、BACE1、PS1、PEN-2、APH1-a蛋白的表達(dá)。結(jié)果:1.新物體識(shí)別實(shí)驗(yàn):AD組相比對(duì)照組,新物體的識(shí)別指數(shù)明顯降低(P0.05);H102組相比AD組,新物體的識(shí)別指數(shù)明顯升高(P0.05);H102組與對(duì)照組相比,無(wú)顯著性差異(P0.05)。2.定位航行實(shí)驗(yàn):AD組相比對(duì)照組,逃避潛伏期延長(zhǎng)(P0.05);H102組相比AD組,逃避潛伏期縮短(P0.05);然后進(jìn)行空間探索實(shí)驗(yàn),AD組相比對(duì)照組,第三象限停留時(shí)間和跨越隱匿平臺(tái)的次數(shù)明顯減少,入水朝向角明顯增大(P0.05);H102組相比AD組,第三象限停留時(shí)間和跨越隱匿平臺(tái)的次數(shù)明顯增加,入水朝向角減小(P0.05);H102組與對(duì)照組相比,均無(wú)顯著性差異(P0.05)。3.免疫組化測(cè)試:AD組小鼠腦內(nèi)BACE1、PS1、PEN-2、APH1-a蛋白表達(dá)較對(duì)照組顯著升高,ADAM10和ADAM17的蛋白表達(dá)相比于對(duì)照組顯著降低(P0.05);H102組Aβ、BACE1、PS1、PEN-2、APH1-a蛋白比AD組顯著降低,ADAM10、ADAM17蛋白表達(dá)比AD組顯著升高(P0.05);H102組與對(duì)照組相比,均無(wú)顯著性差異(P0.05)。4.Western blot檢測(cè):AD組小鼠腦內(nèi)BACE1、PS1、PEN-2、APH1-a蛋白表達(dá)比對(duì)照組增高,ADAM10和ADAM17蛋白的表達(dá)比對(duì)照組減少(P0.05);H102組BACE1、PS1、PEN-2、APH1-a蛋白比AD組顯著降低,ADAM10和ADAM17蛋白的表達(dá)比AD組顯著升高增加(P0.05);H102組與對(duì)照組相比,均無(wú)顯著性差異(P0.05)。5.Aβ蛋白檢測(cè)結(jié)果:免疫組化和ELISA實(shí)驗(yàn)方法檢測(cè)均表明,AD組相比對(duì)照組,Aβ蛋白的表達(dá)顯著升高(P0.05);H102組相比AD組,Aβ蛋白的表達(dá)明顯減少(P0.05);H102組與對(duì)照組相比,無(wú)顯著性差異(P0.05)。結(jié)論:β片層阻斷肽H102可影響AD小鼠腦內(nèi)APP的代謝過(guò)程,減少Aβ、BACE1、PS1、PEN-2、APH1-a蛋白的表達(dá),增加ADAM10、ADAM17蛋白的含量,減少Aβ的生成,H102可以增強(qiáng)AD小鼠的學(xué)習(xí)記憶能力。
[Abstract]:Aim: to observe the effects of 尾 -lamellar blocking peptide H102 on the metabolic secretory enzymes of A 尾 -app and the learning and memory ability of AD mice in APP/PS1 double transgenic AD mice.Method 1: 1.Thirty 6-month-old APP/PS1 double transgenic mice were randomly divided into AD group and H102 group. Fifteen C57BL/6J mice of the same age and the same background were used as control group.By nasal administration, at the same time,The mice in H102 group were given 5 渭 l excipient solution once a day with 5.8 mg/kg)5 渭 l AD and 5 渭 l in control group, and the learning and memory abilities of each group were tested by new object recognition test and Morris water maze test after 30 days of continuous administration.After the behavioral experiment was completed, 10% chloral hydrate was injected intraperitoneally into the anesthetized PBS, the mice were killed, the brain was taken from ice, one side of the brain tissue was embedded in 4% paraformaldehyde prepared in advance for 24 hours, and the embedded tissue was made into sections.Immunohistochemical method was used to detect the expression of ADAM10, ADAM17, BACE1, PS1, PEN-2, APH1-a 尾, and the other half, after separating hippocampus and cortex, stored in liquid nitrogen for a short time, then removed it from the liquid nitrogen and stored it in -80 鈩，

本文編號(hào)：1734095